Nasturtium (Tropaeolum Majus L) Intake and Biochemical Parameters in Pre-diabetic Subjects in Bogota Colombia

Study Overview
=================
Brief Summary
-----------------
Brassicaceae plant family have a high content of bioactive compounds such as e.g. glucosinolates (GSLs) and isothiocyanates (ICTs) associated, recently, with diabetes prevention. This research proposal has the intention of evaluating if the ingestion of freeze-dried nasturtium has a positive effect on the insulin response, lipid profile, oxidative stress biomarkers and gene expression of RESISTIN, GLUT 4, acetyl-CoA carboxylase-a (ACC), fatty acid synthase (FASN), NRF-2, NQO1, SFRXN1, glutathione peroxidase 2 (GPx-2), FOXO1, FOXO3 and FOXO6 in subjects with glucose intolerance.

Detailed Description
-----------------
Epidemiological and animal studies have shown that consumption of fruits and vegetables decreases the risk of chronic diseases. Especially in T2D, the primary prevention is considered as a public health priority. Some studies show an inverse association between vegetables intake and T2D prevention, although the underlying mechanisms and the effect of individual antioxidant compounds are still unclear. Brassicaceae plant family (e.g., broccoli, cabbage, brussels sprouts, watercress, Nasturtium, garden cress, kale, cauliflower), have a high content of bioactive compounds such as e.g. glucosinolates (GSLs), isothiocyanates (ICTs) and polyphenols, associated, recently, with diabetes prevention (18). Human studies in diabetic patients have shown that the consumption of broccoli sprouts powder (BSP) containing high concentration of the GLS sulphoraphane for 4 weeks decreases serum insulin concentration, homoeostasis model assessment of insulin resistance (HOMA-IR), LDL, inflammation and antioxidant markers. Besides sulphoraphane other GLSs have showed a protective effect in diabetes. Recently, mice fed with very high fat diet (VHFD) supplemented with 5% of moringa concentrate rich in benzyl isothiocyanate showed that moringa isothiocyanates (MICs) improved glucose tolerance and insulin signaling; reduced plasma leptin, resistin, cholesterol, IL-1β, TNFα, and lower hepatic glucose-6-phosphatase (G6Pase) gene expression. This evidence suggest that MICs could play a role in T2D prevention probably by inhibiting rate-limiting steps in liver gluconeogenesis resulting in direct or indirect increase in insulin signaling and sensitivity. Recently in vitro studies in human and osteosarcoma hepatoma cells (HepG2 cells) stimulated with benzyl isothiocyanate (BITC) from Tropaeolum majus, show that this compound is able of 1) regulate the insulin signaling pathway, 2) down-regulate the gene and protein expression of gluconeogenic enzymes G6Pase and phosphoenol piruvatokinase (PEPCK) and 3) up-regulate the gene expression of antioxidant and phase II detoxification genes manganese superoxide dismutase (MnSOD), nuclear factor (erythroid derived)-like2 (NRF2), NAD(P)H dehydrogenase (quinone 1) (NQO1) and sulfiredoxin-1 (SFRXN1) (21). According with this evidence BITC could mimics the fasting state, in which insulin stimuli is absent and transcription factors remain in the nucleus modulating gene expression of their target genes, with the advantage of down-regulating gluconeogenesis instead of increase it suggesting that BITC could have a therapeutic role in the prevention or treatment of T2D. Despite of strong evidence of 1) chronic inflammation as an underlying cause of T2D, and 2) high levels of oxidative stress under T2D conditions the use of ITC-rich foods as therapeutics in T2D remains virtually unknown. For that reason this research proposal has the intention of evaluating if the ingestion of freeze-dried nasturtium has a positive effect on the insulin response, lipid profile, oxidative stress biomarkers and gene expression of RESISTIN, GLUT 4, acetyl-CoA carboxylase-a (ACC), fatty acid synthase (FASN), NRF-2, NQO1, SFRXN1, glutathione peroxidase 2 (GPx-2), FOXO1, FOXO3 and FOXO6 in subjects with glucose intolerance.

Official Title
-----------------
Effect of Consumption of a Lyophilized Nasturtium (Tropaeolum Majus L), on the Insulin Response, Lipid Profile, Antioxidant Capacity and Gene Expression in Pre-diabetic Subjects: Bogotá-Colombia

Conditions
-----------------
Pre Diabetes

Intervention / Treatment
-----------------
* Dietary Supplement: Nasturtium (Tropaeolum majus)
* Dietary Supplement: Placebo


Participation Criteria
=================
Eligibility Criteria
-----------------
Inclusion Criteria: Patients aged 25-60 years, with a clinical diagnosis of glucose intolerance for at least six months. Body mass index (BMI) > 30 kg/m2. The BMI will be restricted to > 30 kg/m2 to reduce the variability in the lipid profile parameters, which frequently are normal in subject with normal BMI. Sign informed consent Exclusion Criteria: Severe impairment of cardiac, hepatic or renal function. Gestation or lactation Use of insulin injection Pharmacotherapy with metformin or iDPP4, which alter insulin sensitivity or Glycaemia Consumption of antioxidant or vitamin supplements Consumption of estrogen or vitamin K-antagonists

Ages Eligible for Study
-----------------
Minimum Age: 25 Years
Maximum Age: 60 Years

Sexes Eligible for Study
-----------------
All

Accepts Healthy Volunteers
-----------------
No

Study Plan
=================
How is the study designed?
-----------------

Design Details

Primary Purpose: Prevention
Allocation: Randomized
Intervention Model: Crossover Assignment
Masking: Quadruple


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Placebo Comparator: Placebo (PLC)<br>3g collagen colored with green pigment used in the food industry; Each participant was supplemented with 15 grams of freeze-dried nasturtium leaf powder (NT) or placebo (PLC) per wk during 4 wk. Each participant received 3 envelopes per wk with 5 g NT each or 3 g PLC. In total each participant was given 24 envelopes (12 NT and 12 PLC) packaged in the same way to be indistinguishable from each other. The participants were instructed to take three days at wk one NT or PLC envelope diluted in 300 ml (about 10.14 oz) in chilly water in a bottle provided for the study | Dietary Supplement: Placebo<br>* collagen colored with green pigment used in the food industry<br>|
| Experimental: Nasturtium (NT)<br>5g freeze-dried nasturtium leaf powder ; Each participant was supplemented with 15 grams of freeze-dried nasturtium leaf powder (NT) or placebo (PLC) per wk during 4 wk. Each participant received 3 envelopes per wk with 5 g NT each or 3 g PLC. In total each participant was given 24 envelopes (12 NT and 12 PLC) packaged in the same way to be indistinguishable from each other. The participants were instructed to take three days at wk one NT or PLC envelope diluted in 300 ml (about 10.14 oz) in chilly water in a bottle provided for the study | Dietary Supplement: Nasturtium (Tropaeolum majus)<br>* Freeze dried nasturtium (Edible flower)<br>|

What is the study measuring?
-----------------
Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Glucose response | Fasting and postprandial blood glucose (mmol/L) was measured by the enzymatic colorimetric method (Beckman Coulter, CA, USA) | 4 weeks | 
| Lipid profile | total cholesterol (mmol/L), triglycerides level (mmol/L), High density lipoprotein cholesterol (mmol/L) and Low density lipoprotein cholesterol (mmol/L) were measured by enzymatic colorimetric analysis (Beckman Coulter, CA, USA). | 4 weeks | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| IR-HOMA | Homeostatic model assessment for Insulin Resistance was calculated with the formula (Basal Insulin (μU/L) x fasting blood glucose (mmol/L)/22.5) | 4 weeks | 
| Insulin | Insulin (uU/mL) was measured by chemiluminescent microparticle immunoassay (CMIA) (Abbott, IL, USA). | 4 weeks | 
| Castelli's risk index I and II | CT/HDL-c and LDL-c/HDL-c were calculated as cardiovascular risk factors parameters, at baseline and 4 weeks after treatment. | 4 weeks | 
| Oxidized LDL | OX-LDL (pg/mL). For quantify the concentration of serum oxidized LDL used ELISA kit (Novus Biologicals, USA) | 4 weeks | 
| Atherogenic Coefficient (AC) | non- HDLc)/ HDL-c were calculated as cardiovascular risk factors parameters, at baseline and 4 weeks after treatment. | 4 weeks | 

 Terms related to the study
=================
Keywords Provided by Centre Hospitalier Valida
-----------------
Tropaeolum majus L.