Gene Modulation of NLRP3, IL-1β and TNF-α in Peripheral Blood of Patients With Exogenous Obesity Treated With Berberine
Phase: NA
Conditions: Obesity, Overweight, Berberine
Sponsor: National Polytechnic Institute, Mexico

 
1 
 
 
 
 
NATIONAL POLYTECHNIC INSTITUTE 
 
      School of Medicine 
 
                     SECTION OF GRADUATE STUDIES AND RESEARCH 
 
 
 
 
 
Principal Clinical Research 
 
PhD Modesto Gómez López 
PhD Eleazar Lara Padilla 
 
 
Clinical sub-investigator 
 
PhD Nadia Mabel Pérez Vielma 
 
 Nutritionist María del Refugio Moyetón Hernández 
 
ESM CE-01/7-12-2015 
 
 
Mexico City, 15 August 2015
“Gene modulation of NLRP3, IL-1β and TNF-α in 
peripheral blood of patients with exogenous obesity 
treated with Berberine.” 

 
1 
ABSTRACT 
 
Background: In the treatment of exogenous obesity more than 800 traditional plants are 
used, such is the case of Berberine a phytopharmaceutical. This phytodrug has been 
shown to be anti-obesogenic in the Asian population, however its effect on our population 
is unknown.   Objective: To determine the effects of Berberine on the gene modulation 
of NLRP3, IL-1β and TNF-α in peripheral blood of patients with exogenous obesity. 120 
patients were studied, treated with: hypocaloric nutritional plan, hypocaloric nutritional 
plan and moderate aerobic exercise, hypocaloric nutritional plan, moderate aerobic 
exercise plus Berberine and finally a normo-weight group without treatment. 
Anthropometric measurements, biochemical tests, clinical and gene expression data of 
NLRP3, IL-1β and TNF-α were obtained. The anthropometric measurements, the 
treatment with Berberina obtained, with the exception of ICC, statistically significant 
results, while in the biochemical results the groups 2 and 3 showed similar results, those 
treated with Berberina only the VLDL 
 
 
 
. 
  
 
 
 
 
 
 
 

 
2 
1. INTRODUCCIÓN 
 
Obesity is a systemic, progressive, chronic disease of multifactorial etiology, 
currently the most important and growing public health problem1. 1 This pathological 
condition has been present since remote antiquity, persists and is further increased by 
genetic and environmental factors, and is currently a pandemic with close links to the main 
causes of morbidity and mortality2. 
 
The process of globalization has brought about changes that have modified 
customs and habits, but above all, our lifestyle. This has led to a considerable increase in 
chronic non-communicable diseases, such as obesity3. 
 
According to the Clinical Practice Guideline 'Prevention, diagnosis and treatment 
of overweight and exogenous obesity', as well as NOM-008-SSA3-2017, for the 
comprehensive treatment of overweight and obesity, a personalized nutritional plan and 
physical activity/exercise is the initial basis in the management of this pathology, since 
lifestyle modification is the basis of treatment. The second option for treatment would be 
pharmacotherapy and ultimately bariatric surgery, depending on the Body Mass Index 
(BMI) of each patient4,5. 
 
Despite the advances in the treatment of obesity that Mexico has made in health in 
recent years, this continues to be a major problem to be solved in the country. This 
pathology is due to inadequate eating habits and sedentary lifestyles, which implies 
increasing the efforts of the population/authorities to reduce its prevalence. Therefore, the 
study proposes the implementation of a nutritional plan, moderate aerobic physical 
activity, as well as the use of phytopharmaceuticals as an alternative coadjuvant therapy 
for the management of obesity, from a clinical, biochemical and molecular point of view. 
 
 
 

 
3 
2.  THEORETICAL FRAMEWORK 
 
2.1. 
 Obesity 
““Chronic low-grade systemic inflammatory process due to abnormal secretion of 
cytokines and other inflammatory factors. “6 
  
2.2. 
Definition 
 
Obesity is defined, according to the World Health Organization (WHO), as an 
abnormal or excessive accumulation of fat that can be detrimental to health, although this 
concept has changed over time, from excessive accumulation of body fat, greater than or 
equal to 20% of body weight, greater energy intake, compared to that used, causing an 
increase in body weight/adipose tissue or as an unfavorable health condition, caused by 
a positive energy balance maintained over time, characterized by an excessive increase 
in body fat deposits and body weight 7, 8, 9.  
 
According to the World Health Organization (WHO), obesity is defined as abnormal 
or excessive accumulation of fat that can be harmful to health.  This concept has changed 
over time, from excessive accumulation of body fat, greater than or equal to 20% of body 
weight, greater energy intake, compared to that used, causing an increase in body 
weight/adipose tissue or as an unfavorable health condition, caused by a positive energy 
balance maintained over time, which is characterized by an excessive increase in body 
fat stores and body weight 7, 8, 9. It is not understood how the concept of obesity has 
changed. I suggest just leaving the WHO and the most recent or complete. Separate the 
ideas or definitions with a period. 
 
While NOM 174-SSA1-1998 specifies “a disease characterized by the excess of 
adipose tissue in the body”, on the other hand, the Official Standard for the Treatment of 
Obesity in Mexico 2010 defines it as “a disease characterized by the excess of adipose 
tissue in the body, determined with a BMI equal or greater than 30 kg/m2 and 25 kg/m2 in 
subjects with height below average height ”10, determined with a BMI equal to or greater 

 
4 
than 30 kg/m2 and 25 kg/m2 in subjects with height below the average height “10 , and 
NOM 008-SSA3-2010, for the comprehensive treatment of overweight and obesity, 
defines it as ‘abnormal and excessive accumulation of body fat’. Both, overweight and 
obesity, are accompanied by metabolic alterations that increase the risk of developing 
comorbidities such as arterial hypertension, type 2 diabetes, cardiovascular and 
cerebrovascular diseases, as well as breast, endometrial, colon and prostate neoplasms, 
among others11. Whatever the definition, there are 3 fundamental components that 
coincide: excess body fat, higher caloric intake with lower energy expenditure and weight 
gain. 
 
2.3. 
Epidemiology of Obesity 
 
Obesity has been recognized as a disease by the WHO in 1979 and is considered 
the greatest public health problem; since 1960 the prevalence of obesity has been 
increasing, and our country's transition from infectious diseases and malnutrition to having 
the first place in childhood obesity and the second place in adult obesity in the world, 
associating this pathology with other chronic non-communicable diseases12,13.  
 
In 1998, obesity was classified by the WHO as an epidemic, due to the fact that 
globally, there were more than 1 billion overweight adults and 300 million of these were 
obese. This epidemic is not restricted to developed societies, in fact, its increase is more 
rapid in developing countries, such as Mexico where it faces a double challenge together 
with its malnutrition problems14.  
 
The WHO reported in 2016, that the prevalence (1975 to 2016) of 
overweight/obesity worldwide increased to almost double with more than 1900 million 
adults aged 18 years and older, diagnosed as overweight and of these, more than 650 
million are obese. About 39% of adults are overweight (39% men and 40% women); 
overall, 13% of the world's adult population are obese (11% men and 15% women), in 
both categories the prevalence is higher in women 15. 
 

 
5 
The highest prevalence of overweight and obesity was recorded in the adult 
population in the American continent (overweight: 62% in both sexes; obesity: 26%)12 , 
with the United States of America (USA) in first place, with the highest obesity rate in the 
world, and Mexico in second place with 32%16. 
 
In the specific case of Mexico, data from the OMENT (Mexican Observatory of 
Noncommunicable Diseases) show that the epidemiological situation in the country has 
changed in recent decades17, since malnutrition and infectious diseases were the public 
health problems18. Currently we are a nation with greater obesity in adults, by 2015 a 
report by Obesity Update referred that Mexico had the fastest increase in the world in 
overweight/obesity, for that year Mexico ranked second in adult obesity with 30.7% and it 
was projected that by 2030 it will increase to 39%17.  
 
In 2016 ENSANUT reported that the prevalence of overweight and obesity in 
women increased with respect to 2012, not only in adults but also in girls and adolescents, 
where the greatest increase was in rural areas. In the case of the combined prevalence 
of overweight/obesity in adults over 20 years of age, the data indicate that seven out of 
10 adults suffer from this disease (72.5%), this percentage increased vs. 71.2% in 2012. 
The case of adult women was 75.6%, the increase was higher in rural areas (8.4%) vs. 
urban areas (1.6%). In men, the combined prevalence was 69.4%, with an increase of 
10.5% vs. 201219,20.   
 
When comparing the abdominal phenotype of obesity in adults >20 years of age 
was 76.6% with a higher prevalence in the female sex with 87.7% compared to 65.4% in 
men; the highest proportion of this disease is concentrated in groups from 40 to 79 years 
of age. In this category, the only significant percentage difference with the ENSANUT 
2012 was 4.9 points in the case of the female sex. In other words, 72% of adults continue 
to be overweight or obese 19. Therefore, there is concern that generations born in recent 
decades, because of diseases and health damage caused by obesity, may have lower 
longevity than previous generations13. 
 

 
6 
 
 
 
 
 
 
 
 
 
 
 
 
Figure 1: Comparison of the results of Prevalence of overweight and obesity in adult population in the 
ENSANUT, 2016. 
 
 
 
 
 
 
 
 
 
 
 
 
 
Figure 2: Comparison of the results of Trend of abdominal obesity in adult population in the ENSANUT, 
2016. 
 
2.4. 
Diagnosis and clinical classification 
 
The diagnosis of overweight and obesity should be obtained by anthropometry (weight, 
height, waist and hip, % body and visceral fat), blood biochemistry, medical clinical history, 
as well as dietary clinical history21,22. 
 

 
7 
  
The diagnosis of obesity is obtained by classifying the body mass index (BMI), i.e. a 
person's weight in kilograms divided by the square of the height in meters squared. A 
person with a BMI equal to or greater than 25 is considered overweight and with a BMI 
equal to or greater than 30 is considered obese7.  
 
The Mexican Official Standard 008 SSA3-2016, for the comprehensive treatment of 
overweight and obesity, makes the modification for the Mexican population according to 
short stature which, classifies it as a measurement less than 1.50 meters in women and 
less than 1.60 meters in men10. 
 
 
Table 1. Classification of overweight and obesity according to BMI. 
Classification 
BMI (Kg/m2) 
Classification 
BMI (Kg/m2) 
Low height 
Normal weight  
18.50-24.99 
 
 
Overweight  
> 25 y <29.9 
Overweight  
≥ 23 y < 24.9  
Obesity 
 
Obesity 
≥ 25 
Grade 1 
30 a 34.99 
 
 
Grade 2  
35 a 39.9 
 
 
Grade 3 
≥ 40 
 
 
. Table1. The classification of the degree of obesity using the body mass index as a criterion, distinguishing the following 
degrees referred to in Table 1: Being obesity as BMI equal to or greater than 30, while with short stature that classifies 
it as a measurement less than 1.50 meters in women and less than 1.60 meters in men referred to in Table 1, according 
to the Mexican Official Standard 008 SSA3-2016. Source: WHO and NOM 008. 
 
The clinical classification of obesity is derived in two: exogenous and endogenous23,24,25.  
 
Exogenous: this obesity results from an imbalance between calories consumed and 
calories expended due to a lack of physical activity, i.e., excessive eating and sedentary 
lifestyle; exogenous obesity is currently considered the most prevalent not only in Mexico, 
but in the world18,26.  

 
8 
Endogenous: caused by metabolic alterations, also called intrinsic or secondary18,26,27,28. 
 
Body fat distribution is divided into the following types:  
 
Android (abdominovisceral or visceroportal), there is predominance of adipose tissue in 
the abdominal area, this type is more frequent in males and is strongly related to 
cardiovascular risk. Gynecoid (femoro gluteal), predominates in the lower abdomen, hips 
and buttocks and is more common in women18,29,30. The interest in the genetics associated 
with the obesity phenotype is due to the variability between individuals, where fat 
deposition is controlled by gene expression. 
  
 
 
 
 
 
 
 
 
Figura 
3. 
Distribución 
Androide 
y 
Ginecoide 
en 
Obesidad, 
tomada 
del 
sitio 
web: 
(http://reduvirtualcbs.xoc.uam.mx/obesidad/imagenes/m3_imag1.gif ) 
 
2.5. 
Etiology 
  
Obesity is a multifactorial pathology. Obesity-related habits can be divided into two 
very important fields: genetic and environmental 31,32,33,34. 
 
The most influential factor in obesity is the environmental factor, where poor 
nutrition is fundamental due to the imbalance of energy intake and lack of physical activity. 
Nowadays it is known that there is a direct relationship with energy intake, since currently 
unhealthy eating patterns such as the consumption of sugary drinks or industrialized 
foods, where Mexico has the highest per capita consumption of industrialized beverages, 

 
9 
which are the main causes of malnutrition, and although there have been public policies 
focused on the reduction of this type of products35, in addition to the intervention in 
nutritional labeling, tax increase on foods high in sugars, to mention a few 36, 37, so far they 
have not been able to reduce in a potential way the national obesity prevalence, but in 
addition this prevalence has been increasing alarmingly.  
 
There are other possible causes for the development of overweight and obesity 
that are not modifiable, such as age, genetics, race, ethnicity and sex38. While there are 
other modifiable causes such as sociocultural: family environment, economic income, 
educational level, marital status; biological: number of pregnancies (women); behavioral: 
smoking, alcoholism, depression, anxiety or some other psychopathologies; and those 
related to physical activity: daily activities, exercise or sport39,40. 
 
Among the genetic causes, genes related to the control of energy intake and use 
have been characterized and studied, in addition to their polymorphisms40,41,42,43.  
In addition, the current lifestyle with insufficient sleep, high stress, excessive use 
of electronic devices, among others, are the main causes of weight imbalance in 
Mexicans44. 
2.6. 
Comorbidities associated with obesity. 
Obesity is considered the main risk factor for the development of other 
comorbidities, among which are45,46: 
 
Type 2 diabetes: like obesity, diabetes is more prevalent in women and is more 
prevalent in urban areas than in rural areas47,48,48,49,50. Obesity is related to the causality 
of type 2 diabetes, with 90% of the prevalence attributed to it, due to the lipotoxicity 
inherent to prolonged obesity; thus, almost 10% of the Mexican population lives with 
diabetes, although it is considered that a similar number is undiagnosed. The 
complications of this disease are the leading cause of mortality in Mexico: every year, 
98,000 people die from diabetes worldwide48,49,50,51,52. 
 

 
10 
Arterial hypertension (AHT): it is fully recognized that obesity is a cardiovascular 
risk factor independent of the patient's blood pressure; android obesity is the highest risk 
factor, not only for AHT but also for increasing cardiovascular risk factors and, once again, 
the female sex has a higher prevalence 53,54,55,56,57. 
 
Dyslipidemias: an adult with obesity is up to four times more likely to be diagnosed 
with dyslipidemias, compared to an adult with a healthy weight. In Mexico, the 
measurements recorded in the last 30 years show a trend towards an increase in 
overweight and obesity42,58.  
 
2.7. 
Pathophysiology of obesity 
 
Obesity, according to its origin, can be classified in two: exogenous and endogenous. 
Endogenous obesity is related to endocrinopathies, such as: polycystic ovary syndrome, 
hypothyroidism, hyperinsulinism secondary to pancreatic tumor, Cushing's syndrome, 
hypogonadism, among others, this represents 5 to 10% (of what population?)59, 60 .Obesity 
of exogenous origin is related to epigenetic factors induced by: abuse of high -fat food 
intake, processed foods, diets rich in refined carbohydrates, sugar-sweetened beverages 
and alcohol, which promote weight gain. In addition to inappropriate physical activity 
(sedentary lifestyle).  
 
3. Nutritional plan 
Nutritional management of obesity aims to reduce weight by 5% to 10%; this 
percentage is related to improved health due to a reduction in the comorbidities of this 
pathology; these benefits are greater with greater weight reduction in dyslipidemia, 
hyperglycemia and hypertension, and a reduction of at least 10% in weight is required to 
observe clinical changes, as well as a reduction in mortality of up to 15%. For a correct 
evaluation of the nutritional plan, individual anthropometric, biochemical, clinical and 
dietary data should be taken and a specific nutritional plan should be prepared according 
to the needs and conditions of each patient62. For this it is necessary to determine the 

 
11 
basal and total energy expenditure; with the evaluation the nutritional diagnosis is made 
and the degree of obesity of the patient is known and with this the nutritional plan is 
elaborated.   
3.1 Energy intake 
Energy intake is a homeostatic process of body functioning and balances the amount of 
energy that is stored as fat and its catabolism63; this regulation resides in the 
hypothalamus where the regulation of body composition is integrated with energy intake 
and expenditure64.  
Energy expenditure is the energy used and consists of four components: basal metabolic 
rate, endogenous thermogenesis, physical activity and resting energy expenditure65. 
The basal metabolic rate (BMR) of the individual is the energy expended at complete 
physical and psychological rest, for the correct functioning of all organs and for cellular 
respiration, and accounts for 60%-70% of an individual's total energy expenditure. On the 
other hand, thermogenesis is 10% of total expenditure in sedentary individuals and 
increases in athletes, reaching 50% of total expenditure in elite athletes66, 67. 
3.2  Energy expenditure 
Basal energy expenditure (BEE) is calculated with different formulas, the Harris-Benedict 
formula being the most used, determined as follows: 
TMB Woman = 655 + (9.6 * P) + (1.8 * A) – (4.7 * E) 
TMB Men = 66 + (13.7 * P) + (5 * A) – (6.8 *  E) 
The total energy expenditure (TEE) is then estimated by multiplying the GEB by 
the individual's physical activity. The GET indicates how much is required to cover energy 
needs, the result is complemented with 24-hour recall and weekly food frequency, 
validated by ENSANUT 65.  

 
12 
The calorie prescription that induces a negative balance in obese patients is from: 
Kcal/day= GET or GEB-caloric deficit, this deficit is from 50 to 1000 Kcal to decrease 500-
1,000 g per week66. 
3.3 Macronutrient distribution 
 
CARBOHYDRATES: is the main base of the diet, they are 50% of the total caloric 
value, they are composed of (glucose): carbons (6), hydrogens (12) and oxygens (6), 
carbohydrates are divided into three groups:  
 
Monosaccharides, disaccharides and polysaccharides, the first two are considered 
simple and polysaccharides, complex; of the total carbohydrates consumed it is 
recommended that 90% be complex 67-69.   
 
FATS: constitute up to 30% of the total caloric value of the diet, and their 
composition is carbon, hydrogen and oxygen; the term fats refers to fats and oils in food 
and are a mixture of fatty acids, which are divided into three:  
 
Saturated, monounsaturated and polyunsaturated, and it is recommended that 7%-
9% calories be saturated fatty acids, monounsaturated 15%-18%, and polyunsaturated 
7%-9%: (omega 6 ratio of 10:1 and omega 3 of 5:1), and although it is promoted to ingest 
mono and polyunsaturated fatty acids for their protective effect, currently the intake of fats 
is higher than recommended with an obesogenic effect 69,70. 
 
PROTEINS: like carbohydrates and fats, proteins have carbon, hydrogen, oxygen 
and nitrogen and are determined by the biological value of the protein, which can be of 
low (vegetable origin) or high biological value (animal origin), the importance of this 
macronutrient lies in its functions of growth, cellular repair, to mention a few, and its 
recommended consumption is 20% of the total value69. 
 
 
 

 
13 
3.4  Hypocaloric diet 
 
The hypocaloric diet is the nutritional plan with energy restriction, to generate a negative 
energy balance and reduce body weight. It is recommended in overweight or obese adults, 
where the intake of macronutrients, vitamins and minerals is adequate; if the intake is less 
than 1200 Kcal, it is supplemented to ensure proper nutrition 71.  
 
4. Exercise 
 
The effect of exercise on obesity goes beyond decreasing weight, as it improves 
anthropometry, biochemistry and clinical, and improves mental health, so it is important 
to increase physical activity and exercise for greater energy consumption and, therefore, 
use the body's energy reserve. When implementing exercise should consider the 
characteristics of the individual to be programmed, be pleasant and consistent with their 
lifestyle so that they adapt appropriately.  
 
Studies in different populations where physical activity, exercise and nutritional changes 
were combined obtained good results in the treatment of overweight/obesity72. 
 
4.1 Moderate aerobic exercise 
 
Moderate aerobic exercise 30 minutes a day, at least 3 times a week increases 
lipolysis up to 3 times, since fatty acids are released and used as a source of energy and 
the risk of diseases such as hypertension, diabetes and cancer is reduced, insulin 
sensitivity increases, triglycerides decrease and cardiac output increases73, 74. 
 
5. Adipose tissue 
 
Adipose tissue is very important in obesity67; white adipose tissue (unilocular) stores 
triglycerides in a single compartment68. It is distributed in subcutaneous adipose tissue 

 
14 
and visceral adipose tissue69; adipocytes generate and receive information from the 
environment and are involved in the chronic inflammation of obesity76.  
 
In the pro-inflammatory state associated with obesity, the expansion of adipose tissue 
plays a determining role: adipose tissue increases the production of adipocytokines that 
mediate 
pro- 
and anti-inflammatory and immunological reactions, 
triggering 
pathophysiological processes related to inflammation, such as microvascular 
complications, where pro-inflammatory adipokines are elevated and anti-inflammatory 
ones are decreased75,77. In other words, although individuals are genetically programmed, 
interaction with environmental factors modulates gene expression. 
 
Cellular components of adipose tissue  
 
Adipose tissue is composed of different cell lines: endothelial, mesenchymal, vascular, 
fibroblasts, leukocytes and macrophages, the latter being part of the immune system and 
derived from monocytes formed in the bone marrow77. 
 
In obesity, inflammatory and metabolic pathways are associated with an increase in the 
number of resident macrophages in the adipose tissue78 and can be M1 (classic or pro-
inflammatory) or M2 (alternative or anti-inflammatory)79. It is not only important how many 
macrophages there are in adipose tissue, but also their function; thus, M2 macrophages 
have greater expression of anti-inflammatory genes in lean subjects, whereas in obese 
subjects there are more M1 macrophages with greater release of pro-inflammatory 
adipocytokines77. 
  
Adipose tissue and inflammation  
Exogenous obesity is producing an accumulation of free fatty acids resulting from 
lipogenesis in the form of triglycerides to adipocytes. This accumulation and differentiation 
of mesenchymal stem cells cause hypertrophy and hyperplasia, inflaming the AT, which 
in turn causes activation of the first line of immune defense with specialized cells such as: 

 
15 
macrophages and dendritic cells; which recognize endogenous and exogenous ligands, 
this recognition is performed by Toll-like receptors (TLR's) that activate signaling 
pathways culminating in the translocation of nuclear transcription factor kappa B (NF-kB) 
which plays a fundamental role in inflammation. Because the secretion of inflammatory 
cytokines by activated leukocytes, macrophages, and other tissues are mainly regulated 
by this signaling pathway, this pathway activates inflammasome proteins which is a 
mediator of inflammation, that is, inflammation is promoted by TLR's through the 
production of proinflammatory cytokines and this condition causes an increase in the 
cellular compartments of the subcutaneous adipose tissue, increasing macrophages. 
Such infiltration by macrophages derived from circulating monocytes have the function of 
tissue repair and homeostasis, although due to inflammation such macrophages 
differentiate under the pressure of the tissue microenvironment into M1 macrophages that 
secrete a large number of inflammatory mediators, that is, the synthesis and secretion of 
proinflammatory cytokines are mainly by the macrophage, however, cytokines of the IL-
1β family are exceptional. Unlike the rest of the cytokines, these, too, are dependent on 
NLRP3 inflammasome formation, although these cytokines differ in their initial activation, 
they converge and coincide with the same inflammatory genes, to summarize; the 
interaction between proinflammatory pathways (NF-kB) and cellular response to genomic 
damage, coexist in that microenvironment of the obese AT, where, the deregulated 
activation of the NLRP3 inflammasome contributes in pathological processes, being the 
inflammation characteristic of obesity one of the crucial risk factors since plasma levels of 
certain cytokines such as: NLRP3, IL-1β and TNFα are elevated in obese subjects and 
are related to their body fat and body weight. On the other hand, total body weight loss 
has been associated with a decrease in these cytokines90, 91, 92, 93, 94, 95.   
NLRP3 
The best characterized inflammasome to date is NLRP3, its gene located on chromosome 
1q44, has nine exons; it codes for the 1034 aa cryopyrin protein, the expression of this 
protein is highly regulated and is affected by pro-inflammatory processes and local 
environmental factors. During its activation conformational changes are induced to recruit 
the adaptor protein ACS, and this interacts with procaspase-1 in its inactive form, through 

 
16 
the CARD domain, this domain is present in both proteins. Subsequently, activated 
caspase-1 is responsible for the maturation of pro.IL1β, to obtain the biologically active 
form of IL-1β90, 91, 92, 93, 94, 95. 
Tumor necrosis factor-alpha (TNF-α):  
Obesity-associated inflammation alters the functionality of AT, releasing free fatty acids 
into the circulation, resulting from TNF-α-favored lipolysis, initiating inflammatory 
signaling. TNF-α is an adipose tissue protein that is elevated in obesity and contributes to 
insulin resistance. Its gene is located on chromosome 6p21.3; this gene has a promoter 
region, four exons and three introns. This cytokine is synthesized by macrophages, T 
lymphocytes, endothelial cells among other cells, a membrane-bound precursor that 
exerts an inflammatory biological role locally and depending on the interaction with other 
cells, and another soluble one that exerts its inflammatory functions at a distance from the 
cells that synthesize it, the membrane-bound form gives rise to the soluble form of this 
cytosine (REVIEW HOW TNF IS SYNTHETIZED), the latter is made up of 233 aa and has 
a molecular weight of 26KDa while TNF-α is made up of 157 aa and has a molecular 
weight of 17 Kda; Finally, the active form of TNF-α consists of a 51 KDa homotrimer, which 
can dissociate at low concentrations, thus losing its biological activity96, 97. 
IL-1β: 
Like TNF-α, IL-1β initiates inflammatory signaling through its receptors. IL-1β is 
linked in the pathogenesis of DT2, as it promotes insulin resistance and induces apoptosis 
and alters the function of β-cells of the islets of Langerhans of the pancreas because of 
inflammation. In addition, IL-1β promotes the synthesis of TNF-α, which decreases insulin 
signaling. Saturated fatty acids released into the bloodstream cause macrophages to 
decrease AMPK (Adenosine Monophosphate-Activated Protein Kinase)98. 
Specifically, IL-1β synthesis is given in the mRNA coding for pro-IL-1β and is 
subsequently translated by polysomes in the cytosol until activated by IL-1β converting 
enzyme (ICE). This molecule can be translocated to facilitate its localization at the cell 
membrane, the converting enzyme is translated in the endoplasmic reticulum as an 

 
17 
inactive precursor, remains in the cytosol and requires two internal anchors (also by ICE) 
to form an enzymatically active heterodimer. Then, two heterodimers form a tetramer in 
association with two pro-IL-1 β molecules and anchoring occurs. After anchoring the 17kD 
IL-1 β is secreted into the extracellular milieu through a membrane channel. IL-1 β (amino 
acids 1-116) can be found both inside and outside the cell. A small amount of pro-IL-1 β 
can be transported into intact cells, presumably through the same channel. In the absence 
of ICE, pro-IL-1ß can be activated by extracellular proteases. 
In the specific case of obesity, fatty acid secretion tips the balance towards 
increased recruitment of M1, secreting pro-inflammatory factors, including TNF-α and IL-
1β, among others. This accumulation of M1 macrophages in adipose tissue is the most 
important source of IL-1β and caspase-1 through the formation of the NLRP 
inflammasome3 98, 99, 100. 
5. 
Treatment 
According to the Clinical Practice Guide 'Prevention, diagnosis and treatment of 
overweight and exogenous obesity', the initial evaluation identifies individuals with 
overweight/obesity, as well as those at risk of suffering from them, who can benefit from 
weight loss. In the study of obesity, anthropometry, blood biochemistry, clinical and diet, 
as well as economic, are evaluated to have a detailed overview of the individual and how 
to make a comprehensive intervention5. 
 
The Mexican Official Standard NOM-008-SSA3-2017 for the comprehensive 
treatment of overweight and obesity refers to the set of actions based on the complete 
and individualized study of the patient with overweight or obesity, and includes medical, 
nutritional, psychological, rehabilitation, physical activity and, where appropriate, surgical, 
aimed at changing the lifestyle, reducing health risks, comorbidities and improving the 
quality of life of the patient4. In addition, to treat an overweight/obese individual, the 
following must be available: Clinical scale with stadiometer and equipment to treat 
overweight/obese individuals, anthropometric tape, and those established by applicable 
regulations. 

 
18 
Because the diagnosis of obesity by BMI can be imprecise, other variables in 
addition to the weight and height of the individual must be taken into account; thus, the 
Edmonton Obesity Staging System (EOSS) identifies comorbidities and functional 
limitation in obesity, to have precise therapeutic objectives in individuals with a BMI 
greater than or equal to 30.101 
 
The consensus of the Spanish Society for the study of Obesity 2016 for the 
prevention, diagnosis and treatment of obesity, the treatment guidelines according to the 
BMI of the obese individual as another form of intervention in their treatment 102.Currently, 
Mexico does not have effective treatments against obesity. This is evident since ENSANUT MC 
2016 reported an increase compared to ENSANUT 2012, placing Mexico in second place 
worldwide in adult obesity and first place in childhood obesity, indicating that strategies to reduce 
this pathology have not been successful; therefore, it is necessary to adapt comprehensive and 
multidisciplinary treatments that contribute to the intervention in this pathology. 
 
The objective of using drugs is to reduce complications caused by obesity and increase 
quality of life; thus, the greatest advantage in reducing risk factors is obtained by losing 5-
10% of weight in 6 months and attenuating health risks, especially cardiometabolic ones. 
This indicates that drugs for overweight/obesity are prescribed when there is evidence of 
non-response to diet therapy, physical activity and exercise, and that in 3-6 months there 
has been no loss of >5% of weight in individuals with a BMI of 30 kg/m2 with/without 
comorbidities or in those with a BMI greater than 27 with one or more comorbidities, a 
drug is used when the aforementioned criteria are met106. 
 
There are various drugs used for weight loss such as orlistat, and recently the European 
Medicines Agency approved two new drugs: liraglutide and the combination of bupropion 
with naltrexone (2015), with side effects: nausea-vomiting, with liraglutide: headaches, dry 
mouth, nausea and dizziness, with bupropion/naltrexone: CNS alterations in the case of 
anorexigenics due to its interaction with catecholamines, which is why the implementation 
of alternative treatments with fewer adverse effects is required102, 106, 107. 
 
 

 
19 
Use of phytopharmaceuticals 
 
The General Health Law in article 224 defines phytopharmaceuticals or herbal medicine 
as: 
“Products made with plant material or a derivative thereof, whose main ingredient is the 
aerial or underground part of a plant (extracts and tinctures), as well as juices, resins, fatty 
and essential oils, presented in pharmaceutical form, whose therapeutic efficacy and 
safety has been scientifically confirmed in national or international literature”108. Currently, 
plants are used in infusion and in capsules, syrups or extracts109. While the regulation of 
health supplies defines: 
 
“Herbal medicines, in addition to containing plant material, may add excipients and 
additives in their formulation. Those that are associated with isolated and chemically 
defined active ingredients, or those proposed as injectables, are not considered herbal 
medicines. In the formulation of a herbal medicine, narcotic or psychotropic substances 
of synthetic origin, or mixtures with allopathic medicines, procaine, ephedrine, yohimbine, 
chaparral, germanium, animal or human hormones or other substances that contain 
hormonal or antihormonal activity or any other that represents a risk to health may not be 
included”110. In this way, the quality of a phytopharmaceutical and its use as an effective 
medicine for a specific disease can be guaranteed. It must also be supported by medical 
research that endorses its results111. 
 
The difference between a phytopharmaceutical and a chemical drug is that the former 
contains as an active ingredient a plant preparation instead of a synthesized chemical 
substance112, 113. 
 
Berberine 
 
Berberine is an alkaloid of Rhizoma coptidis, Cortex pellodendri and Hydrastis 
canadensis, and is used as an over-the-counter medicine in China to treat infectious 

 
20 
diarrhea, inflammation, diabetes, non-alcoholic fatty liver disease, dyslipidemia, 
cardiovascular diseases, etc.114, 115, 116. 
 
Berberine has activity against bacteria, viruses, fungi, protozoa, helminths and chlamydia, 
with in vitro and clinical phase results. Berberine is antidiarrheal by inhibiting peristalsis 
and, probably, by its intestinal antisecretory action. Different clinical studies show that 
Berberine is effective in treating acute diarrhea caused by enterotoxins from E. coli, Vibrio 
cholerae, among others. It is also more effective than ranitidine for duodenal ulcers. In th e 
cardiovascular system it has positive inotropic, negative chronotropic, anti-arrhythmic and 
vasodilatory action. The decrease in platelet aggregation and thromboxane B2 could be 
involved in its anti-ischemic effect; it also has cytostatic activity in vivo and in vitro, 
stimulates the secretion of bile and bilirubin, improves the symptoms of chronic 
cholecystitis and normalizes tyramine concentration in patients with liver cirrhosis110, 117, 
118. 
 
It also has anti-tumor effects in many types of cancer, since it suppresses proliferation by 
inducing apoptosis, inhibiting metastasis, and has very low or no toxicity in healthy tissue 
and is protective in the liver against chemically induced hepatotoxicity111,112,113,117,118,119. 
 
Berberine and its interaction with obesity and inflammation 
 
The main component of Rhizoma coptidis is Berberine, it is biologically active and is an 
over-the-counter drug with multiple properties, among which anti-inflammation stands out, 
reducing IL-1β, TNF-α, iNOS, ICAM-1, IL-6 and the activation of NF-κB in vivo. Berberine 
reduces inflammation via TLR4, since it inhibits the activation of SRC and cell mobility by 
TLR in macrophages stimulated with LPS; in addition, it inhibits the expression of TNF-α, 
IL-6, TLR 2, TLR 4 and TLR 9 in early-phase sepsis in an animal model114,120. 
 
The suppression of inflammation by Berberine is a complex phenomenon, the NF-κB 
pathway plays an important role, since TLR-NF-κB mediate inflammation and could be 
targeted by Berberine to induce anti-inflammatory activity. Inflammatory stimuli include 

 
21 
TNF-α, in the NF-κB pathway; these factors could be inhibited by Berberine, which would 
negatively regulate the NF-κB pathway with a decrease in pro-inflammatory cytokines by 
macrophages and suppression of inflammation 121, 122, 123. In addition, Berberine and 
metformin promote glucose metabolism by stimulating glycolysis, which is probably due 
to the inhibition of the mitochondrial respiratory chain complex, independent of AMPK 
activation, meaning that this phytopharmaceutical has a hypoglycemic effect124. Berberine 
protects the heart from ischemia/reperfusion injury, since it reduced myocardial injury by 
decreasing myocardial autophagy and apoptosis via AMPK/mTOR in animals125. 
 
Berberine interactions with inhibitory and activating genes of inflammation. 
 
In silico Berberine shows a strong binding with TLR4 receptors and other anti- and 
pro-inflammatory genes, coinciding with results reported in vitro and experimental model. 
 
 
 
Figure 10. Image taken from the reference: Di Pierro Francesco, the at. Clinical role of a fixed combination 
of standardized Berberis aristata and Silybum marianum extracts in diabetic and hypercholesterolemic 
patients intolerant to statins. Diabetes, Metabolic Syndrome and Obesity: Targets and Therapy 2015:8. 89-
96 

 
22 
6.  Justification 
 
The inflammatory process in chronic non-communicable diseases, such as obesity, alters 
the patient's health, and also promotes tissue damage and insulin resistance, which in 
turn induce type 2 diabetes or hypertension, to name a few, so the immunomodulatory 
effect of Berberine may be a viable alternative to reduce this process in exogenous 
obesity. 
 
7. Problem statement  
 
Obesity is highly prevalent in our country and is associated with chronic inflammation and 
high morbidity and mortality. Despite scientific advances in its study, the mechanisms that 
lead to this state are not clear. In obese individuals, prone to comorbidities, the use of 
additional intervention is justified, and although there are various treatments, the 
development of new products or combinations of them is increasingly important. Animal 
studies indicate that Berberine inhibits the production of pro-inflammatory cytokines (IL-
1β and TNF-α) although they do not report on NLRP3. 
The clinical/molecular evaluation of Berberine in obese individuals will determine whether 
the phytopharmaceutical has therapeutic potential. 
 
8. Research Question 
 
What is the effect of Berberine on the gene expression of IL-1β, TNF-α and NLRP3, in 
peripheral blood cells in individuals with exogenous obesity? 
 

 
23 
9. Objectives 
 
General objective: 
To determine the effects of Berberine on gene expression of cytokines IL-1β, TNF-α 
and NLRP3 in individuals with exogenous obesity. 
 
Specific Objectives: 
1. To form groups diagnosed with exogenous obesity. 
2. To evaluate anthropometric parameters 
3. To evaluate biochemical parameters 
4. To quantify the expression (mRNA) of NLRP3, IL-1β and TNF-α 
5. To establish the differences in gene expression between individuals with exogenous 
obesity and control subjects. 
 
10. Hypothesis 
 
Berberine inhibits gene expression of NLRP3, IL-1β and TNF-α in individuals with 
exogenous obesity. 
 
 
 
 
 

 
24 
11. Material and methods 
 
Type of study: 
An experimental, analytical, longitudinal, prospective study of comparative groups was 
carried out. 
Study subjects 
Ninety patients with type I and II obesity will be selected, as well as 30 control subjects 
(healthy adults), who agreed to be donors of a peripheral blood sample, 2 tubes (1: 
biochemistry, 2: gene expression) channeled to the Obesity Program unit 
(Comprehensive Medical Service, ESM, IPN), with the following selection criteria. 
12.1. Selection criteria 
Inclusion 
Human peripheral blood obtained from adult donors with the following characteristics: 
• Patients with BMI ≥ 30kg/m2 
• With risk factors for type 2 diabetes (history of diabetic parents or siblings, sedentary 
habits) 
• Fasting blood glucose ≤ 126 mg/dL or glycosylated hemoglobin < 6.5% 
• Controlled arterial hypertension 
• >18 years of age 
• Accept and sign the informed consent 
No inclusion 
• Hypersensitivity to the components of the formula 
• History of hypoglycemic episodes 
• History of obesity of endogenous origin 
• History of serious psychological, psychiatric, eating or neurological disorders 

 
25 
• History of hyperthyroidism, heart disease, diabetes, liver or kidney failure, glaucoma, 
epilepsy 
• Pregnancy or lactation 
Exclusion criteria 
• Pregnancy 
• Diabetics 
• Presenting any serious adverse effect 
• Patients who decide to withdraw from the protocol 
Elimination 
• 
Samples contaminated during processing 
• 
Lysed blood samples 
• 
Coagulated blood simple. 
Variable Description 
 
Independent Variable 
Berberine 
Category: quantitative. 
Measurement scale: ratio. 
Measurement unit: 500 mg with excipient q.s. 
Operational definition: the tested dose of Berberine was 500 mg, given to patients for 3 
months. 
 
Dependent 
Gene expression of IL-1β, TNF-α and NLRP3. 
Category: quantitative. 
Measurement scale: discrete, continuous. 
Measurement unit: arbitrary units. 

 
26 
Operational 
definition: 
Cytokines were determined with commercial 
qPCR 
equipment/reagents, and their arbitrary units/relative expression were determined by a 
specific standard curve for each one. 
 
Observation method: open. 
 
Sampling method: non-probabilistic sampling of consecutive comparative groups that 
met the selection criteria, until reaching the sample size that constituted the study groups 
(patients with exogenous obesity). 
 
Operational description of the study: Berberine has been provided by the Medix 
laboratory, in 500 mg capsules, to treat patients for 3 months. 
 
Study groups 
 
The individuals will be distributed into four groups: 
 
• Group 1. Individuals with exogenous obesity: nutritional plan intervention for lifestyle 
change. 
• Group 2. Individuals with exogenous obesity: nutritional plan intervention, moderate 
controlled aerobic exercise for lifestyle change. 
• Group 3. Individuals with exogenous obesity: nutritional plan intervention, moderate 
aerobic exercise and Berberine phytopharmaceutical. With biweekly sessions for 3 
months. 
• Group 4. Normal-weight individuals without intervention. 
 
Blood samples were taken after informed consent from the participants, they were 
informed of the intervention and the group to which they belonged, they were also taken 
at the end of the study. 
 
 

 
27 
Procedure 
 
In the selection, obtaining information, anthropometry and blood biochemistry, the 
subjects of the study will be asked for authorization and their participation will be 
requested after a detailed explanation of the risks and benefits of the protocol, obtaining 
signed informed consent. 
 
Obtaining the samples 
 
The universe of the study will be individuals from 19 to 55 years of age with exogenous 
obesity; they will be recruited in the Obesity Projects area of the Postgraduate Studies 
and Research section of the Higher School of Medicine of the National Polytechnic 
Institute, referred by the Comprehensive Medical Service. 
 
Each participant will be asked in writing to enter the study, each one was identified, and a 
clinical history was prepared with anthropometric measurements: weight, percentage of 
body fat, percentage of visceral fat, height, BMI. Peripheral blood will also be taken for 
blood biochemistry and messenger RNA extraction and qPCR tests to determine gene 
expression of NLRP3, IL-1β and TNF-α. 
 
Biochemical tests. 
 
Peripheral blood samples will be evaluated by an external laboratory. 
Blood samples will be collected at the beginning and three months after treatment. 
Participants were called at 7 am, after fasting for at least 8 hours, and the sample was 
collected in Vacutainer tubes (BD, USA), added with ethylenediaminetetraacetic acid 
(EDTA) to prevent coagulation. 4 mL of blood was taken from everyone; from there, 1 mL 
of Trizol (Sigma-Aldrich, USA) was added to 2 2 mL tubes with 100 µL per tube. The blood 
was vortexed for 30 s and stored at -70 °C until processing. The rest of the blood was 
stored at -20 °C. 
 

 
28 
Anthropometric evaluation 
 
The somatometry of each participant will be determined by wearing light clothing and 
without shoes: weight, height, % body fat, % visceral fat. 
 
Height 
It will be defined as the distance between the vertex and the support plane. 
• Instrument: Stadiometer (Seca mod. 208). 
• The measurement will be expressed in cm with a precision of 1 mm. 
The subject stood up straight, with the heels together and supported on the back stop and 
so that the inner edge of the feet formed an angle of 60 degrees. The buttocks and the 
upper part of the back were in contact with the vertical table of the stadiometer. The 
subject was instructed to breathe deeply without getting up and maintaining the position 
of the head. The horizontal platform of the stadiometer was slowly lowered until it 
contacted the head of the subject, applying gentle pressure to minimize the effect of the 
hair and placing the head in the Frankfort plane. 
 
Weight, % body fat and % visceral fat 
• Instrument: Body composition analyzer (InBody770). 
• The measurement will be expressed in Kg, with a precision of 0.1 kg. 
It will be determined with the body analyzer, programmed with the required parameters 
(age, sex, height, among others). The subject stood in the center of the body analyzer 
platform, distributing the weight equally on both legs, without the body being in contact 
with anything around it, and placed his feet and hands on the equipment electrodes with 
his arms extended on both sides of the body, until the record was displayed on the screen 
and monitor. 
 
Body Mass Index 
 
The body mass index (BMI), also known as the Quetelet index, is an index of adiposity 
and obesity, as it is directly related to the percentage of body fat. The body mass index 

 
29 
was calculated by dividing the weight in kilograms by the square of the height in meters. 
This index has the advantage that its components (weight and height) can be easily 
measured. The nutritional status of participants was classified based on international 
anthropometric indicators, according to the WHO. 
 
Nutritional Plan 
 
There is no unanimity regarding the “ideal diet” because everyone has different 
anthropometry, blood biochemistry, clinical features, habits, economy, customs, 
pathologies, family/social interactions, which make standardization impossible. Therefore, 
based on the initial assessment, the following were prescribed: 
 
“Classic” low-calorie diet: Recommended in most guides and consensus, it establishes a 
deficit of 500 to 1,000 calories per day, and a balanced distribution of nutrients: 45-55% 
carbohydrates, 15-25% protein, 25-35% total fat, and 20-40 g of fiber 1, 41. 
 
“Low-calorie” Mediterranean diet: This is the model supported by SEEDO because it 
represents this balanced and healthy approach, with a low intake of saturated and trans 
fatty acids and added sugars, and a high consumption of vegetable fiber and 
monounsaturated fatty acids. Its health benefits, including mortality, are clearly 
established in the literature. 
 
High-protein diet: Based on the satiating effect and less loss of lean mass. The intake is 
increased to >1.2 g/kg/day, with a minimum intake of 90 g daily. 
 
Nutritional Management 
 
The nutritional management plan in this project will be aimed at improving the lifestyle and 
quality of life of patients, as well as reducing their body weight. This intervention will be 
done according to the nutritional management for obesity developed by nutritionists from 
INCMNSZ. 

 
30 
The goals of the nutritional plan were to establish healthy eating habits with a 10-15% 
reduction in body weight during treatment. 
 
Nutritional Consultations 
 
The consultations will be biweekly to evaluate body weight, anthropometry and adherence 
to the nutritional regimen, in addition to monitoring the modification of eating habits. 
 
Clinical-nutritional history 
 
In this clinical history, the lifestyle of the patients will be evaluate, the activities from the 
time they will wake up until bedtime will be questioned, their physical activities, allergies 
or intolerances, eating habits such as: meal times, tastes, preferences, quantities, type of 
food preparation, economic amount allocated, places of food, if they ate with electronic 
distractions, among others, to know the needs of each participant and adequately 
prescribe the food plan. 
 
Food frequency 
 
A food frequency survey from ENSANUT 2012 will be applied to learn about eating habits 
and determine the consumption of energy, important macro and micronutrients. The same 
survey analyzes and reports the results when capturing the data. With the evaluation of 
the diet and the anthropometric, biochemical, clinical, dietary and economic indicators, a 
nutritional diagnosis and the degree of obesity of each patient were established. 
 
Energy expenditure 
 
After the diagnosis, the prescription of the individualized food plan was prepared. The 
basal energy expenditure (BEE) was calculated with the Harris Benedict formula, and 
the current body weight. 
 

 
31 
While the total energy expenditure (TEE) was estimated from the physical activity of 
everyone by multiplying the BEE by the physical activity factor, according to the category 
of each one. 
 
Energy prescription 
 
A negative balance was induced to decrease 1kg of body weight per week, reducing from 
500 to 1,000 Kcal per day. The caloric deficit was determined from the TEE for their current 
weight, the daily energy intake of the participants where the number of calories was 
calculated having the total energy expenditure minus 20% to obtain homogeneity in all 
participants. 
 
Dietary distribution and composition. 
 
The nutritional distribution with 50% carbohydrates was recommended, 10% simple 
carbohydrates, from fruit consumption, while the distribution with 30% fats was 
recommended, 10% saturated fats, and finally the distribution with 20% protein was half 
of animal origin and half of vegetable origin. The meal plans were adapted according to 
the requirements of each participant. 
 
Each meal plan contained sample menus and an additional food exchange brochure, 
which contained the food groups and their equivalents, depending on whether the 
participants' weight decreased or the number of portions of each food group was 
maintained, as well as the calories of each nutritional plan. 
 
The number of mealtimes was distributed in 5: 3 main meals (breakfast, lunch and dinner) 
and two additional snacks (morning and afternoon). 
 
The lists of the Mexican System of Equivalents and the Institutional Program for the 
Prevention and Control of Diabetes Mellitus, IMSS, were used to prepare a list of portions 

 
32 
of the most consumed/common foods that were organized into food groups. The objective 
was to facilitate the participants' practice of new eating habits. 
 
Anthropometric Evaluation 
It allowed us to identify alterations in the nutritional status of each participant, as well as the 
associated risks; the anthropometric variables that were evaluated at the beginning of the 
program, every 15 days and at the end allowed us to assess the changes in the participants 
beyond weight loss, granting nutritional plans according to the specific needs for their 
anthropometric, biochemical, clinical, dietary, economic assessment and according to their 
family history. 
Phytopharmaceutical 
Therapeutic indications 
A bottle of Berberine of 500 mg with excipient cbp of 102 tablets each was given, as well as 
a new bottle at each consultation and the participants handed over the empty bottles or with 
leftover pills. The way they should ingest them was 3 tablets: 3 times a day before each 
meal. 
Treatment adherence log 
The patients will give a treatment adherence log to indicate the day and time they took the 
tablets, as well as if they did not take them. 
Adverse effects log 
The patients will give a log in which they should indicate if they had any adverse events, to 
report them. 
Controlled Aerobic Exercise 
Controlled aerobic exercise on a treadmill, supervised by a sports physician, the frequency 
with which participants carried out their sessions was 3 times a week for 30 minutes each, 
at 60-70% of maximum heart rate. Due to the sedentary lifestyle of the patients, coupled 

 
33 
with obesity and its complications, an ‘app’ was installed on the participants’ cell phones 
(with the exception of two: one did not have the device and one could not read or write, so 
they were provided with a semi-automated pedometer so that all participants had the same 
intervention), all were asked if they exercised, which was denied and their usual physical 
activity was minimal; in the initial phase of the protocol, their usual physical activity was 
determined (using the ‘app’), which they were instructed to increase, with a goal of 10,000 
steps or 150 min per week. 
Gene expression 
Extraction of total RNA 
Blood samples will be lysed, and the material was extracted with TRIzol®Reagent, a mixture 
of guanidine isocyanate and phenol-chloroform, following the manufacturer's instructions. 
Once isolated, the total RNA will be suspended in RNase-free water to prevent its 
degradation and stored at -80°C. 140 µL of peripheral blood were used. 
Extraction technique 
Sample lysis will be carried out by adding one milliliter of TRIzol reagent and allowing it to 
incubate for 10 minutes. 
1) Separation phase. Phenol-chloroform (0.2 mL per sample) was added, and the mixture 
was then centrifuged. Three phases were separated: a lower organic phase with the 
proteins, an interface with the DNA, and a transparent upper aqueous phase with the RNA. 
This fraction was transferred to a new tube to precipitate the RNA. 
2) RNA precipitation by adding 0.5 mL of isopropyl alcohol. Centrifugation at 12,000g for 10 
min at 4 C occurs, and the precipitated RNA forms a pellet or adheres to the walls and base 
of the tube. 
3) RNA washing. This is done with 1 mL of 75% alcohol and centrifugation at 7500g for 5 
minutes. The pellet is dried by vacuum centrifugation (spin vac pump) until the alcohol 
evaporates. 

 
34 
4) Re-suspension of total RNA. The samples are dissolved in RNAse-free water, treated 
with 0.1% diethylpyrocarbonate (DEPC) (nuclease inhibitor). The integrity of the tRNA is 
checked on an agarose gel. Quantification of total extracted RNA 
Sample concentration and purity will be measured by spectrophotometry (NanoDrop2000™, 
Thermo Fisher Scientific Inc., Waltham, Massachusetts, USA). Samples with a purity index 
(at 260/280nm) of 1.80-2.0 were considered adequate, indicating the absence of 
contamination. 
Complementary DNA synthesis by reverse transcription 
 
cDNA amplification will be performed using the “Transcriptor First Strand cDNA” kit for Rt-
PCR from Roche (Roche Diagnostics GmbH, Mannheim, Germany), according to the 
manufacturer’s instructions. 500 µg of the cDNA sample was used in 20 µl volume. For the 
reaction, the following was done: The cDNA was synthesized from the tRNA of the samples 
and with the Invitrogen Roche kit (CA USA), in the following reaction mixture: 
 
1) H2O mili Q - 12.5 µL (the volume varies) 
2) Buffer 10x MMLV – 2.5 µL 
3) ‘Random primer’ - 0.5 µL 
4) DTT 0.1 M—2.5 µL 
5) dNTPs mix –1 µL 
6) MMLV RT 
 
The volume of the mixture is adjusted to 25 µL and the final product after the reaction in the 
conventional thermocycler (MasterCycler, Eppendorf) for 1 hour and stored at -20ºC. 
 
Real-time polymerase chain reaction, qRT-PCR 
 
Procedure to determine the relative mRNA expression of genes of interest, using probes 
from the human transcriptome library (Roche, Library), LightCycler nano thermal cycler and 
TaqMan reaction mix (Roche Diagnostics). Oligosequences (sense and antisense) were 

 
35 
designed with ProbeFinder software version 2.45 (http://www.universalprobelibrary.com). 
The reaction mix was prepared according to the manufacturer's protocol with 5 µl of cDNA 
of the appropriate dilution, in 10 µl. The assay included a standard curve of 5 serial dilutions 
for each gene, and samples were normalized with the endogenous control β-actin. Each 
sample was performed in triplicate and data were analyzed with LightCycler nano software. 
 
The thermocycler was programmed with the following scheme: preincubation for one cycle 
of 10 minutes at 95°C; the reaction for 45 consecutive cycles (denaturation at 95°C for 10 
seconds, annealing at 60°C for 30 seconds and extension at 72°C for 1 second) and the 
final incubation at 40°C for 30 seconds. 
 
Table 5. Oligonucleotide sequences used to determine gene expression. 
Gen 
Proteins 
Sequences 
Concentration 
NLRP3 
Proteína 
3 
que 
contien
dominios NACHT, LRR y PYD
F: 5´ GCA AGA CTT TGA CAA CAT GC3´ 
R: 3´CAC CTG TTG TGC AAT CTG AAG5´ 
10nM 
TNF-α 
Factor de necrosis tumoral alf F: 5´ CAGCCTCTTCTCCTTCCTGAT3´ 
R: 3´GCCAGAGGGCTGATTAGAGA5´ 
10nM 
IL-1β 
Interleucina 1 beta 
F: 5´ TTG GGT AAT TTT TGG GAT CTT AC3
R: 3´CTG TCC TGC GTG TTG AAA GA5´ 
10nM 
Β-actina 
Gen constitutivo 
F: 5´ AGC CAC ATC GCT CAG ACA C3´ 
R: 3´GCC CAA TAC GAC CAA ATC C5´ 
10nM 
 
Statistical analysis 
The values of each determination are expressed as the mean ± SE. The differences 
between the groups were calculated with ANOVA. Differences between the groups will be 
determined by using Student’s T test. In all tests, a value less than or equal to 0.05 was 
considered significant. 
Ethical aspects 
The study will be of lower than minimum risk, since no blood was obtained from the 
participants in quantities greater than the limit established by the Regulations for 
Research on Human Beings of the General Health Law. The project was evaluated, 

 
36 
approved and accepted by the Research and Ethics Committee of the Higher School of 
Medicine of the National Polytechnic Institute. 
The study was carried out in accordance with the ethical principles: 
• 
 Declaration of Helsinki of the WMA - Ethical principles for medical research involving 
human beings126. 
• 
 Written and signed informed consent as stipulated in article 21 of the General Health 
Regulations on Health Research 127. 
• 
 As stipulated in article 17 of the General Health Regulations on Health Research, 
this project is classified as minimal risk127. 
• 
 The protocol was carried out in compliance with the recommendations established 
in NOM-087-ECOL-SSA1-2002, which addresses matters related to Environmental 
Protection and Environmental Health; Biological-infectious Hazardous Waste128. 
Material Resources 
The project will develop in the Molecular Biology Laboratory and the Obesity Project, 
which have the infrastructure for the protocol. The material, consumables and reagents 
were provided by the responsible researchers, while the Berberine was donated by Medix 
laboratories. 
Financial Resources 
The financial support was provided by the researchers' own resources and by the 
institution. As for the phytopharmaceutical, it was provided by Medix Laboratories. 
 
 
 
 
 

 
37 
Bibliographic references 
 
1. Dávila-Torres J et al. Panorama de la obesidad en México. Rev Med Inst Mex 
Seguro Soc. 2015;53 (2):240-9. 
2. Rodríguez Scull1,  Lidia. Obesidad: fisiología, etiopatogenia y fisiopatología. Rev 
Cubana Endocrinol v.14 n.2 Ciudad de la Habana Mayo-ago. 2003. 
3. Ayuso Peraza, G. Castillo León, M. Globalización y nostalgia. Cambios en la 
alimentación de familias yucatecas. Estud.soc vol.27 no.50 México jul./dic. 2017. 
4. Secretaría de Salud.  Norma Oficial Mexicana NOM-008-SSA3-2017, Tratamiento 
integral del sobrepeso y la obesidad. Diario Oficial de la Federación, 25 de enero 
de 2017.  
5. Barrera-Cruz A et al. Guía para el control del sobrepeso y la obesidad. Rev Med 
Inst Mex Seguro Soc. 2013;51(3):344-57. 
6. Leonie K. Heilbronn, L. V. (2008). Adipose tissue macrophages, low grade 
inflammation and insulin resistance in human obesity. Current Pharmaceutical 
Design, 14(12), 1225-1230. doi:10.2174/138161208784246153 
7. WHO. Obesidad y sobrepeso. World Health Organization; [sede web]. 2015 
[actualizado octubre 2017; citado 12 de Octubre de 2017]; Disponible en: 
http://www.who.int/mediacentre/factsheets/fs311/es/ 
8. Amancio Chassin O. Ortigoza Ramírez, J. Durante Montie, I. [sede web] (2007). El 
ejercicio actual de la Medicina. Obesidad, [sin fecha de actualización; citado 02 de 
Octubre de 2017] 
9. Salinas-Martínez AM, M.-Q. A. (2014). Estimación de Sobrepeso y Obesidad en 
preescolares Normativa nacional e internacional. Revista Médica del Instituto 
Mexicano del Seguro Social, 52(1), S26-S23. 
10. NOM-008-SSA3-2010. Norma Oficial Mexicana NOM-008-SSA3-2010, para el 
tratamiento integral de la obesidad. 
11. Secretaría de Salud.  Norma Oficial Mexicana NOM-008-SSA3-2010, Tratamiento 
integral del sobrepeso y la obesidad. Diario Oficial de la Federación, 06 de Julio 
de 
2010. 
Disponible 
en: 
http://www.dof.gob.mx/nota_detalle.php?codigo=5154226&fecha=04/08/2010 

 
38 
12. Abdullah S AI-Goblan, M. A.-A. (2014). Mecanism linking diabetes mellitus and 
obesity. Dove Press Journal: Diabetes, Metabolic Syndrome and Obesity: Targets 
and Therapy, 2014:7, 587 - 591. doi:http://dx.doi.org/10.2147/DMSO.S67400 
13. Rivera, J.A. y cols. Obesidad en México. Recomendaciones para una política de 
estado, México: UNAM, Dirección General de Publicaciones y Fomento Editorial, 
2012. Rivera, J.A. y cols. Obesidad en México. Recomendaciones para una 
política de estado, México: UNAM, Dirección General de Publicaciones y Fomento 
Editorial, 2012. 
14. Moreno G Manuel. Definición y clasificación de la obesidad. Rev. Med. Clin. 
Condes 
- 
2012; 
23(2) 
124-128. 
Disponible 
en: 
https://ac.els-
cdn.com/S0716864012702882/1-s2.0-
S0716864012702882main.pdf?_tid=e23087ad-efff-4a2e-9c5b-
e3fbe3de27e8&acdnat=1529541123_f0865e6e30dcfd2e2c67295abcf94e35 
15. WHO. Obesidad y sobrepeso, datos y cifras, [sede web]. 2018 [actualizado 2015; 
citado 16 de Febrero de 2018]; Disponible en: http://www.who.int/es/news-
room/fact-sheets/detail/obesity-and-overweight 
16. OECD Obesity Update [sede web] 2017 [actualizado 2018, citado 02 de 
septiembre 
2018]. 
Disponible 
en: 
http://www.oecd.org/health/obesity-
update.htm 
17. OMENT Observatorio Mexicano de Enfermedades No Trasmisibles. Mexico ocupa 
el 2do lugar en obesidad en adultos según la OCDE. [sede web] 2017 
[actualizado 12 de julio 2017, citado 18 de agosto 2018]. Disponible en: 
http://oment.uanl.mx/mexico-ocupa-el-2o-lugar-en-obesidad-en-adultos-segun-la-
ocde/ 
18. Poveda, Duran, Manuel. Polo Ruiz-Tovar, Jaime. González, González. Juan. 
Conmorbilidades de la obesidad. En: Poveda, Duran, Manuel. Polo Ruiz-Tovar, 
Jaime. González, González. Juan. Principios de obesidad mórbida. Madrid. 
DYKINSON, S.L.2017. p. 29-30. 
19. OMENT Observatorio Mexicano de Enfermedades No Trasmisibles. Cifras de 
Sobrepeso y Obesidad en México-ENSANUT MC 2016. [sede web] 2016 
[actualizado 20 de diciembre 2016, citado 18 de agosto 2018]. Disponible en: 

 
39 
http://oment.uanl.mx/cifras-de-sobrepeso-y-obesidad-en-mexico-ensanut-mc-
2016/ 
20. WHO. Organización Mundial de la Salud, Estadísticas Sanitarias Mundiales 
[sede web].2012 
[citado el 24 de enero de 2017]. Disponible en: 
http://apps.who.int/iris/bitstream/10665/44858/1/9789243564449_spa.pdf?ua
=1 
21. González Garrido J, Ceballos Reyes G, Mélendez Bolaina E. Obesidad: más que 
un problema de peso. La Ciencia y el Hombre. 2010, 23(2). 
22. Manrique M el at. Diagnóstico, evaluación y tratamiento no farmacológico del 
paciente con sobrepeso u obesidad. Statement about diagnosis assessment ano 
non pharmacological treatment of obesity. Rev Méd Chile 2009; 137: 963-971. 
Disponible: 
https://scielo.conicyt.cl/scielo.php?script=sci_arttext&pid=S0034-
98872009000700016. 
23. Moreno, M. Definición y clasificación de la obesidad. Rev. Med. Clin. Condes. 
2012; 23 (2) 124-128. 
24. Youdim A. Obesity. Univadis [Internet]. 2013 [citado el 17 de diciembre 2016]. 
Disponible en: 
http://www.univadis.mx/msd-manual/54/Trastornos- 
nutricionales/Obesidad-y-sindrome-metabolico/Obesidad#t-v886569_es 
25. Secretaría 
de Salud. Sistema Nacional de Vigilancia Epidemiológica. 
Epidemiología. Sistema único de información. 2010; 27(43):1-5. 
26. Arturo Zárate, Lourdes Basurto Acevedo, Renata P. Saucedo García. La obesidad: 
Conceptos actuales sobre fisiopátogenia y tratamiento. Unidad de Investigación 
Médica en Enfermedades Endocrinas, Hospital de Especialidades, Centro Médico 
Nacional: Fac. Medic.; marzo-abril 2001 [citado 22 agosto 2017] volumen 44. 
27.  Breu F, Guggenbichler S, Wollmann J. Consenso SEEDO 2007. Para la 
evaluación del sobrepeso y la obesidad y el establecimiento de criterios de 
intervención terapéutica. Rev. Española Obes. 2007;7–48. 
28. Marbella J. Piñera Tamés. Caracterización clínica y comorbilidades asociadas. 
Efectividad de una intervención grupal vs individual: Madrid 2014; 26. 
29. Navarro MC, Ortega T. Plantas medicinales para el sobrepeso. Madrid: INFITO - 
Editorial Complutense; 2009. 

 
40 
30. Bray GA. Medical consequences of obesity. J Clin Endocrinol Metab. 2004; 89: 
2583-2589. 
31. ADA. American Diabetes Association.2006. Standards of Medical Care in Diabetes–
2006[Sede web] [Sin fecha de actualización; citado 03 de Julio de 2018]; Disponible 
en: http://care.diabetesjournals.org/content/29/suppl_1/s4 
32. Silvestri Eliana. Aspectos psicológicos de la obesidad. Importancia de su 
identificación y abordaje dentro de un enfoque interdisciplinario. INST. MED. INT. 
2005; 4(2); 1-36. 
33. Ravenna, Máximo. Una delgada línea. entre el exceso y la medida. Fac. Medic. 
2004. 6;105-111.  
34. Orlando Horacio. Obesidad: una carrera de regularidad contra el sobrepeso. Clarín 
Buenos Aires. SAOTA. Marzo 2005 
35. SS. Secretaria de Salud. Sobrepeso y obesidad, factores de riesgos para 
desarrollar diabetes. [Sede web] [Fecha de actualización 24 de julio de 2018; 
citado 
29 
de 
agosto 
de 
2018]; 
Disponible 
en: 
https://www.gob.mx/salud/articulos/sobrepeso-y-obesidad-factores-de-riesgos-
para-desarrollar-diabetes?idiom=es 
36. INSP. Instituto Nacional de Salud Pública. Etiquetado Nutrimental (GDA). . [Sede 
web] [Sin fecha de actualización; citado 31 de agosto de 2018]; Disponible en: 
https://www.insp.mx/epppo/blog/4680-etiquetado.html 
37. Fernando Rodríguez R., Ximen Palma L., Ángela Romo B., Daniela Escobar B., 
Bárbara Aragú G., Luis Espinoza O., Norman McMillan L. y Jorge Gálvez C. 
Hábitos alimentarios, actividad física y nivel socioeconómico en estudiantes 
universitarios de Chile. Nutr. Hosp. vol.28 no.2 Madrid mar./abr. 2013 disponible 
en: 
http://scielo.isciii.es/scielo.php?script=sci_arttext&pid=S0212-
16112013000200024 
38. WHO. Datos sobre obesidad; [Sede web]. Internacional: WHO; [sin fecha de 
actualización; 
citado 
12 
de 
Octubre 
de 
2017]; 
Disponible 
en: 
http://www.who.int/features/factfiles/obesity/facts/es/index5.html 
39. Secretaría de Salud. Estrategia Nacional Para La Prevención y El Control Del 
Sobrepeso, La Obesidad Y La Diabetes. México D.F. 2013. 

 
41 
40. Barquera al et. Obesidad en México: epidemiología y políticas de salud para su 
control y prevención Gaceta Médica de México. 2010;146:397-407 disponible en: 
https://www.anmm.org.mx/GMM/2010/n6/38_vol_146_n6.pdf 
41. Mancipe-Navarrete JA, G.-V. S.-B. (2015). Efectividad de las intervenciones 
educativas realizadas en América Latina para la prevención del sobrepeso y 
obesidad infantil en niños escolares de 6 a 17 años: una revisión sistemática. 
Nutriciòn Hospitalaria, 2015:31(1), 102-114. 
42. Juan Ángel Rivera Dommarco. Obesidad en México: recomendaciones para una 
política de Estado. 1° edición, México: Publicaciones fomento editorial; 
2013.Barabási AL. Network medicine from obesity to the "diseasome". N Engl J 
Med. 2007; 357:404-407 
43. Peralta-Romero JJ, G.-Z. J.-V. (2014). Genética de la obesidad infantil. Revista 
Médica del Instituto Mexicano del Seguro Social, 52, s78-s87 
44. SS. Secretaria de Salud. La estrategia Nacional para la prevención y el control del 
sobrepeso, la obesidad y la diabetes. [Sede web] [fecha de actualización Mayo 
2017; 
citado 
31 
de 
agosto 
de 
2018]; 
Disponible 
en: 
https://www.gob.mx/cms/uploads/attachment/file/225245/2_Estrategia_nacional_
para_la_prevencion_y_control_del_sobrepeso_-_DGPS.pdf 
45. Arteaga L. Antonio. Overweight and obesity as an universal health problem. REV. 
MED. CLIN. CONDES. 2012; 23(2): 145-153. 
46. ENSANUT. México, Encuesta Nacional de Salud y Nutrición a Medio Camino, 
[sede web]. 2017 [fecha de actualización 06 de abril 2017; citado 12 de Diciembre 
de 
2017]; 
Disponible en: 
https://www.gob.mx/salud/documentos/encuesta-
nacional-de-salud-y-nutricion-de-medio-camino-2016 
47. Freedman DS, Khan LK, Serdula MK, Galuska DA, Dietz WH. Trends and 
correlates of class 3 obesity in the United States from 1990 through 2000. JAMA 
2002; 288: 1758-1761.  
48. Antonio Arteaga l. El sobrepeso y la obesidad como un problema de salud. Rev. 
Med. Clin. Condes. 2012; 23 (2) 145-153 
49. Siani A, Cappuccio FP, Barba G, Trevisan M, Farinaro E, Lacone R, Russo O, 
Russo P, Mancini M, Strazzullo P. The relationship of waist circumference to blood 

 
42 
pressure: the Olivetti Heart Study. Am J Hypertens. [sede web].2002 [sin fecha de 
actualización; 
citado 
21 
de 
Agosto 
de 
2017]; 
Disponible 
en: 
https://www.ncbi.nlm.nih.gov/pubmed/12219872  
50. FMD. Federación Mexicana de Diabetes. Obesidad y Diabetes, mancuerna 
peligrosa para México. [sede web]. 2018 [actualizado 21 de febrero de 2018; citado 
02 de Octubre de 2017]; Disponible en: http://fmdiabetes.org/obesidad-diabetes-
mancuerna-peligrosa-mexico/ 
51. Manzur F. Alvear C. Alayón A. Adipocitos, obesidad visceral, inflamación y 
enfermedad cardiovascular Adipocytes, visceral obesity, inflammation and 
cardiovascular disease. Rev Colomb Cardiol 2010; 17: 207-213 
52. Parikh NI, Pencina MJ, Wang TJ, Lanier KJ, Fox CS, D'Agostino RB, Vasan RS. 
Increasing Trends in Incidence of Overweight and Obesity over 5 Decades. Am J 
Med. 2007 Mar;120(3):242-50. 
53. Josep Redona,b y Empar Lurbea,c. Hipertensión arterial y obesidad. Med Clin. 
2007;129(17):655-7. 
54. C.M. López de Fez, M.T. Gaztelu, T. Rubio, A. Castaño. Mecanismos de 
hipertensión en obesidad. An. Sist. Sanit. Navar. 2004, Vol. 27, Nº 2:211-219. 
55. Guihovany Alberto García Casilimasa, Daniel Augusto Martinb, María Alejandra 
Martínezb, Claudia Rocío Merchánb, Carol Anne Mayorgab y Andrés Felipe 
Barragánb. Fisiopatología de la hipertensión arterial secundaria a obesidad. 
Pathophysiology of hypertension secondary to obesity. Arch Cardiol Mex 
2017;87:336-44.  
56. De la Figuera von Wichmann, Obesidad e hipertensión arterial. Hipertens Riesgo 
Vasc 2000;17:414-6. 
57. FMD. Federación Mexicana de Diabetes. Dislipidemia en aumento por obesidad. 
[sede web]. 2017 [actualizado 18 de Noviembre de 2015; citado 30 de Agosto de 
2018]; 
Disponible 
en: 
http://fmdiabetes.org/dislipidemia-en-aumento-por-
obesidad/ 
58. Moreno, M. Definición y clasificación de la obesidad. Rev. Med. Clin. Condes. 
2012; 23 (2) 124-128. 

 
43 
59. Youdim A. Obesity. Univadis [sede web]. 2018 [sin fecha de actualización; 
citado el 21 de diciembre 2017]. Disponible en: http://www.univadis.mx/msd-
manual/54/Trastornos-nutricionales/Obesidad-y-sindrome-
metabolico/Obesidad#t-v886569_es  
60. WHO. Organización Mundial de la Salud. Estrategia mundial sobre régimen 
alimentario, actividad física y salud [sede web] 2018 [citado  02 de Septiembre de 
2018]. 
Disponible 
en: 
http://www.who.int/dietphysicalactivity/factsheet_inactivity/es/ 
61. Bulló Bonet M. La leptina en la regulación del balance energético. Nutr Hosp 2002; 
17: 42-8. 
62. Fruh SM. Obesity: Risk factors, complications, and strategies for sustainable long-
term weight management. J Am Assoc Nurse Pract. 2017 Oct; 29 (S1): S3-
S14. Doi: 10.1002 / 2327-6924.12510.  
63. Lenard, N.R. Berthoud, H.R. Central and peripheral regulation of food intake and 
physical activity: pathways and genes. Obesity. 2008. 16 Suppl 3, S11-22 
64. Esteves de Oliveira FC, de Mello Cruz AC, Gonçalves Oliveira C, Rodrigues 
Ferreira Cruz AC, Mayumi Nakajima V, Bressan J. Gasto energético de adultos 
brasileños saludables: una comparación de métodos. Nutr Hosp. 2008; 23:554-61. 
65. Vargas Z,M. Lancheros P,L. Barrera P,M. GASTO ENERGÉTICO EN REPOSO 
Y COMPOSICIÓN CORPORAL EN ADULTOS. Energy expenditure in repose 
related to body composition in adults. Rev. Fac. Med., Volumen 59, Número 1, p. 
43-58, 2011. ISSN electrónico 2357-3848. ISSN impreso 0120-0011. 
66. Obregón Perea; M. Papel de la termogénesis. [sede web]. [sin fecha de 
actualización octubre 2017; citado 12 de Octubre de 2018]; Disponible en: 
http://www.analesranf.com/index.php/mono/article/view/577/594 
67. Manzur F. Alvear C. Alayón A. Adipocitos, obesidad visceral, inflamación y 
enfermedad cardiovascular Adipocytes, visceral obesity, inflammation and 
cardiovascular disease. Rev Colomb Cardiol 2010; 17: 207-213 
68. UNAM. Tejido adiposo. [Sede web].2010. [Sin fecha de actualización; citado 23 de 
Octubre 
de 
2017]; 
Disponible 
en: 

 
44 
http://histologiaunam.mx/descargas/ensenanza/portal_recursos_linea/apuntes/teji
do_adiposo_2010.pdf 
69. FAO. Nutrición humana en el mundo en desarrollo. Macronutrientes: 
carbohidratos, 
grasas 
y 
proteínas. 
Capitulo 
9. 
Disponible 
en:  
http://www.fao.org/tempref/docrep/fao/005/w0073s/W0073S01.pdf 
70. Nakata Y. Web-based intervention to promote weight-loss maintenance using an 
activity monitor: A randomized controlled trial. Prev Med Rep. 2019 Mar 
4;14:100839. doi: 10.1016/j.pmedr.2019.100839. eCollection 2019 Jun. 
71. Alvarez-Alvarez I, et al. Dieta mediterranea hipocalorica y factores de riesgo 
cardiovascular: analisis transversal de PREDIMED-Plus. Rev Esp Cardiol. 2018. 
https://doi.org/10.1016/j.recesp.2018.08.007  
72. Jorquera C. Ejercicio, Obesidad y Sindrome Metabólico. Exercise, Obesity And 
Metabolic Syndrome. Revista Médica Clínica Las Condes. Volume 23, Issue 
3, May 2012, Pages 227-235. 
73. Roldán A, Rendón S. PROPUESTA DE PRESCRIPCIÓN DEL EJERCICIO EN 
OBESOS. Revista Politécnica ISSN 1900-2351, Año 9, Número 16, Páginas 75-
84, 2013. 
74. Aguilar Cordero el at. Programas de actividad física para reducir sobrepeso y 
obesidad en niños y adolescentes; revisión sistemática. Physical activity 
programmes to reduce overweight and obesity in children and adolescents; a 
systematic review. Nutr. Hosp. vol.30 no.4 Madrid oct. 2014 
75. Martos-Moreno G, Argente J. Bases Moleculares de la Obesidad. Ciclo de 
Conferencias. XXXVII Congreso de la Sociedad de Endocrinología Pediátrica 
Española, Oviedo 2005. 
76. Sánchez N, J. López Z.D. Pinzón D, O. Sepúlveda A, J. Adipocinas y síndrome 
metabólico: múltiples facetas de un proceso fisiopatológico complejo Adipokines 
and metabolic syndrome: multiple aspects of a complex pathophysiological 
process. Rev Colomb Cardiol 2010; 17: 167-176. 
77. Meriño,M. Briones,L. Palma,V. Herlitz,K. Escudero, C.  Rol de los receptores de 
adenosina en la interacción adipocito-macrófago durante la obesidad. Role of 

 
45 
adenosine receptors in the adipocyte-macrophage interaction during obesity. 
Endocrinol Diabetes Nutr 2017;64:317-27 
78. Blancas-Flores,G. Almanza-Pérez,J. López-Roa,R. Alarcón-Aguilar,F. García-
Macedo,R. Cruz,M. La obesidad como un proceso inflamatorio. Obesity as an 
inflammatory process. Bol. Med. Hosp. Infant. Mex.2010;. 67:.2. 
79. Fernández Regueras, M.  Almeida Parra, J. Pastor Encinas, I. Orfao, A. Díaz, M. 
Pérez Nieto, M. Laso Guzmán, M. PAPEL DE LA DIFERENCIACIÓN M1/M2 DEL 
SISTEMA MONOCITO/MACRÓFAGO INDUCIDA POR EL CONSUMO DE 
ALCOHOL. Congresos XXXIV Congreso Nacional de las Sociedad Española de 
Medicina Interna (SEMI). XXIX Congreso de la Sociedad Andaluza de Medicina 
Interna (SADEMI). Disponible en: http://www.revclinesp.es/es/congresos/xxxiv-
congreso-nacional-las-sociedad/8/sesion/varios-posters-/849/papel-de-la-
diferenciacin-m1-m2/8665/ 
80. Sarjeant K, Stephens J. Adipogenesis. Cold Spring Harb Perspect Biol. 2012 Sep; 
4(9): 
a008417. 
Disponible 
en: 
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3428766/  
81. Rojas-Gomez et al. Nuevos enfoques moleculares en la regulación de la 
adipogénesis. El papel de la conexina 43.  Archivos Venezolanos de Farmacología 
y Terapéutica [en linea] 2016, 35 [Fecha de consulta: 20 de mayo de 2019] 
Disponible en:<http://www.redalyc.org/articulo.oa?id=55949908002> ISSN 0798-
0264  
82. Aus TariqAli et al. Adipocyte and adipogenesis. European Journal of Cell Biology, 
Volume 
92, 
Issues 
6–7, June–July 
2013, 
Pages 
229-236. 
https://doi.org/10.1016/j.ejcb.2013.06.001. 
83. Espinosa 
Pedroza. 
Lípidos: 
clasificación de 
los 
lípidos, 
unidad 4 
http://www.fmvz.unam.mx/fmvz/p_estudios/apuntes_bioquimica/Unidad_4.pdf 
84. Martínez Sáman el at. Los ácidos grasos y la lipotoxicidad: implicaciones 
metabólicas. Fatty acids and lipotoxicity: metabolic implications. Rev. Fac. Med. 
(Méx.) vol.56 no.1 Ciudad 
de 
México ene./feb. 2013. 
Disponible 
en: 
http://www.scielo.org.mx/scielo.php?pid=S002617422013000500002&script=sci_arttext 

 
46 
85. Angosto, M. C. 2015. Lipogénesis de” novo” y termogénesis. Monografías de la 
Real Academia Nacional de Farmacia. 
86. Pérez 
Mayorga 
EL 
ADIPOCITO 
COMO 
ÓRGANO 
ENDOCRINO. 
IMPLICACIONES FISIOPATOLÓGICAS Y TERAPÉUTICAS. REVISTA 15 (2): 
225-242, 2007 
87. Carvajal C. Lipoproteínas: metabolismo y lipoproteínas aterogénicas. Med. leg. 
Costa Rica vol.31 n.2 Heredia Sep./Dec. 2014. 
88. Romero 
Aguilar 
el 
at. 
CUERPOS 
LIPÍDICOS: 
ORGANELOS 
METABÓLICAMENTE ACTIVOS. Revista de Educación Bioquímica (REB) 
35(4):115-124, 2016. 
89. DiPilato el at. The Role of PDE3B Phosphorylation in the Inhibition of Lipolysis by 
Insulin. Mol Cell Biol. 2015 Aug; 35(16): 2752–2760. 10.1128/MCB.00422-15. 
90. Hernández López. Activación y regulación del inflamasoma NLRP3 en las 
enfermedades infecciosas. Iatreia Vol. 25 (4): 380-390, octubre-diciembre 2012.  
91. Buelvas Jiménez. Regulación del inflamasoma NLRP3: bioquímica y más allá de 
ella. IATREIA Vol 28(2): 170-180, abril, junio 2015. 
92. Abderrazaka A. el at. NLRP3 inflammasome: From a danger signal sensor to a 
regulatory node of oxidative stress and inflammatory diseases. Redox Biology. 
Volume 4, April 2015, Pages 296-307. 
93. Kawana N. Molecular Network of NLRP3 Inflammasome Activation-Responsive 
Genes in a Human Monocyte Cell Line. Austin J Clin Immunol. 2014;1(4): 1017. 
94. Coll C. el at. A small-molecule inhibitor of the NLRP3 inflammasome for the 
treatment of inflammatory diseases. Nature Medicine volume21, pages248–
255 (2015) 
95. Yi Chen el at. The Cancer Prevention, Anti-Inflammatory and Anti-Oxidation of 
Bioactive Phytochemicals Targeting the TLR4 Signaling Pathway.  Int. J. Mol. 
Sci. 2018, 19(9), 2729; https://doi.org/10.3390/ijms19092729 
96. Gutiérrez-Rodelo. Mecanismos Moleculares de la Resistencia a la Insulina: Una 
Actualización. Gac Med Mex. 2017;153:214-28. 

 
47 
97. Fragoso el at. El factor de necrosis tumoral α (TNF-α) en las enfermedades 
autoinmunes (EA): biología molecular y genética. Gaceta medica de Mexico. 2014; 
150:333-44. 
98. Cieślak M et al. Role of pro-inflammatory cytokines of pancreatic islets and 
prospects of elaboration of new methods for the diabetes treatment. Acta Biochim 
Pol. 2015;62(1):15-21. Epub 2015 .Mar 18. 
99. Vélez-Castrillón et al. Bases moleculares de la familia de la interleuquina-1. 
Revista colombiana de reumatología vol. 11 no. 1, marzo 2004, pp. 11-3. 
100. 
Kang YE, Kim JM, Joung KH, Lee JH, You BR, Choi MJ, et al. (2016) The 
Roles of Adipokines, Proinflammatory Cytokines, and Adipose Tissue 
Macrophages in Obesity-Associated Insulin Resistance in Modest Obesity and 
Early 
Metabolic 
Dysfunction. 
PLoS 
ONE 
11(4): 
e0154003. 
https://doi.org/10.1371/journal.pone.0154003. 
101. 
Marcela Rodríguez–Flores. Diagnóstico de la obesidad más allá de índice 
de masa corporal. Salud pública Méx vol.56 no.4 Cuernavaca jul./ago. 2014. 
102. 
Lecube A et al. PREVENCION, DIAGNÓSTICO Y TRATAMIENTO DE LA 
OBESIDAD. 
POSICIONAMIENTO 
SEEDO 
2016. 
Disponible 
en: 
https://www.seedo.es/images/site/ConsensoSEEDO2016.pdf 
103. 
Michelle Mocarski, Ye Tian, B. Gabriel Smolarz, John McAna, Albert 
Crawford. Use of International Classification of Diseases, Ninth Revision Codes for 
Obesity: Trends in the United States from an Electronic Health Record-Derived 
Database. Popul Health Manag. 2018 Jun 1; 21(3): 222–230. Published online 
2018 Jun 1. doi: 10.1089/pop.2017.0092 
104. 
Secretaria de Salud. Centro Nacional de Programas Preventivos y Control 
del Enfermedades. Declaratoria de emergencia. 2018. Disponible en: 
http://www.cenaprece.salud.gob.mx/programas/interior/emergencias/declaratoria
s_emergencia.html 
105. 
Secretaria de Salud. Estrategia Nacional para la prevención y control del 
sobrepeso, 
la 
obesidad 
y 
la 
diabetes. 
Disponible 
en: 
https://www.gob.mx/cms/uploads/attachment/file/276108/estrategia_sobrepeso_d
iabetes_obesidad.pdf 
106. 
Khera R, Murad M, Chandar AK, et al. Association of Pharmacological 
Treatments for Obesity With Weight Loss and Adverse Events: A Systematic 
Review 
and 
Meta-analysis. 
JAMA. 
2016;315(22):2424-2434. 
doi:10.1001/jama.2016.7602.  
107. 
Londoño-Lemos. Tratamiento farmacológico contra la obesidad. Rev. 
Colomb. Cienc. Quím. Farm., Vol. 41 (2), 217-261, 2012 

 
48 
108. 
OMS. Productos herbarios [internet]. International WHO. Disponible en 
http://www.who.int/topics/traditional_medicine/definitions/es/. 
109. 
Gómez Castellanos, J. Rubén, El ambiente regulatorio de los medicamentos 
herbolarios en México. Antecedentes, situación actual y perspectivas al año 2025. 
Boletín Latinoamericano y del Caribe de Plantas Medicinales y Aromáticas 
[internet] 2009, 8 (Sin mes): [Fecha de consulta: 29 de agosto de 2017] Disponible 
en: http://www.redalyc.org/articulo.oa?id=85680107> ISSN 0717-7917 
110. 
RIS. 
Reglamento 
de 
insumos para 
la 
salud. 
Disponible en 
http://www.salud.gob.mx/unidades/cdi/nom/compi/ris.html 
111. 
 Bertha Sola. Fitofármacos que tan efectivo son. 11 de abril de 2016. 
Disponible en:  http://www.cronica.com.mx/notas/2012/611102.html 
112. 
Victoria Hall Ramirez, Ma Soledad Quesada Morúa, Milania Rocha Palma. 
Obesidad Fisiología y Tratamiento. Centro nacional de Medicamentos. Costa Rica, 
Diciembre 2002. 4:49-72. 
113. 
José Antonio García Del Pozo Y Marta Oliva Álvarez Martínez. Plantas 
medicinales en el tratamiento de la obesidad. OFFARM OCTUBRE 2002: VOL 21 
NÚM 9: 132-144. 
114. 
Yang L, Li Z, Haiyan S, 1 and Guang J. 1Update on Berberine in 
Nonalcoholic Fatty Liver Disease. Evid Based Complement Alternat Med. 2013; 
2013: 308134 
115. 
Jin W et al. Coptidis Rhizoma: a comprehensive review of its traditional uses, 
botany, phytochemistry, pharmacology and toxicology. Pharm Biol. 2019; 57(1): 
193–225. 
116. 
Di Pierro Francesco, Iaele Bellone, Giuliana Rapacioli, Pietro Putignano. 
Clinical role of a fixed combination of standardized Berberis aristata and Silybum 
marianum extracts in diabetic and hypercholesterolemic patients intolerant to 
statins. Diabetes, Metabolic Syndrome and Obesity: Targets and Therapy 2015:8. 
89-96. 
117. 
Wei Wei, Hongmin Zhao, Aili Wang, Ming Sui, Kun Liang, Haiyun Deng, 
Yukun Ma, Yajuan Zhang, Hongxiu Zhang, Yuanyuan Guan. A clinical study on the 

 
49 
short-term effect of berberine in comparison to metformin on the metabolic 
characteristics of women with polycystic ovary síndrome. EJE. 2012. 166; 99-105.  
118. 
 Ma. Ester Risco. Plantas medicinales y drogas vegetales. OFFARM. 
Diciembre 2012. Volumen 21;11. 157-158 
119. 
Tung‑Yueh Chuang, Hsiao‑Li Wu, Jie Min, Michael Diamond, Ricardo Azziz 
and Yen‑Hao Chen. Berberine regulates the protein expression of multiple 
tumorigenesis‑related genes in hepatocellular carcinoma cell lines. Bio.Med 
central. 2017. 17:59  
120. 
Chung-Yi C, Chiu-Li K , Chi-Ming L. The Cancer Prevention, Anti-
Inflammatory and Anti-Oxidation of Bioactive Phytochemicals Targeting the TLR4 
Signaling Pathway. Int J Mol Sci. 2018 Sep; 19(9): 2729. 
121. 
Mancini, S. J., White, A. D., Bijland, S., Rutherford, C., Graham, D., Richter, 
E. A. Salt, I. P. (2017). Activation of AMP-activated protein kinase rapidly 
suppresses multiple pro-inflammatory pathways in adipocytes including IL-1 
receptor-associated 
kinase-4 
phosphorylation. Molecular 
and 
cellular 
endocrinology, 440, 44–56. doi:10.1016/j.mce.2016.11.010 
122. 
Sujitha S1, Dinesh P1, Rasool M2. Berberine modulates ASK1 signaling 
mediated through TLR4/TRAF2 via upregulation of miR-23a. Toxicol Appl 
Pharmacol. 2018 Nov 15;359:34-46. doi: 10.1016/j.taap.2018.09.017.  
123. 
Ma X, Chen Z, Wang L, Wang G, Wang Z, Dong X, Wen B and Zhang Z 
(2018) The Pathogenesis of Diabetes Mellitus by Oxidative Stress and 
Inflammation: Its Inhibition by Berberine. Front. Pharmacol. 9:782. doi: 
10.3389/fphar.2018.00782 
124. 
Miao Xu, Yuanyuan Xiao, Jun Yin, Wolin Hou, Xueying Yu, Li Shen, Fang 
Liu, Li Wei, Weiping Jia. Berberine Promotes Glucose Consumption Independently 
of 
AMP-Activated 
Protein 
Kinase 
Activation. 
PLoS 
One. 2014; 9(7): 
e103702. Published online 2014 Jul 29. doi: 10.1371/journal.pone.0103702 
125. 
Qing Y, Dong X, Hongli L, Yanhui L. Berberine promoted myocardial 
protection of postoperative patients through regulating myocardial autophagy. 
Biomed 
Pharmacother. 2018 
Sep;105:1050-1053. 
doi: 
10.1016/j.biopha.2018.06.088. 

 
50 
126. 
Asociacion médica Mundial [sede Web]. Declaración de Helsinki de la AMM 
- Principios éticos para las investigaciones médicas en seres humanos; 2019- 
[actualizada el 21 de Marzo de 2017; acceso 22 de enero de 2017]. Disponible 
en: https://www.wma.net/es/policies-post/declaracion-de-helsinki-de-la-amm-
principios-eticos-para-las-investigaciones-medicas-en-seres-humanos/ 
127. 
Secretaria de Salud [sede Web]. REGLAMENTO de la Ley General de 
Salud en Materia de Investigación para la Salud; 2019 - [sin fecha de actualización; 
acceso 
22 
de 
enero 
de 
2017]. 
Disponible 
en: 
http://www.salud.gob.mx/unidades/cdi/nom/compi/rlgsmis.html 
128. 
Secretaria de Salud [sede Web]. NORMA Oficial Mexicana NOM-087-
ECOL-SSA1-2002, Protección ambiental - Salud ambiental - Residuos peligrosos 
biológico-infecciosos - Clasificación y especificaciones de manejo 2019 - [sin fecha 
de 
actualización; acceso 
22 
de 
enero 
de 
2017]. 
Disponible en: 
http://www.salud.gob.mx/unidades/cdi/nom/087ecolssa.html 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

 
51