Acute Inflammatory and Metabolic Effect of High Fructose Intake
Condition: Inflammatory Response; Metabolic Disturbance
Study type: Interventional

Summary: 
      Fructose consumption is associated with the development of metabolic diseases and low-grade
      inflammation. However, the acute effect of a single meal rich in fructose on the metabolic
      and inflammatory response is not fully understood. This study will to evaluate the acute
      metabolic and inflammatory effect caused by a meal containing fructose overload. This will be
      a three-arm crossover, randomized, double-blind clinical trial.

      Participants will undergo the three interventions for random order: (i) standardized meal
      plus sucrose overload; (ii) standardized meal plus glucose overload; (iii) standardized meal
      plus fructose overload. During the washout period (7 to 21 days), the subjects will
      instructed to maintain their usual eating behavior and physical activity.

      On the day of each intervention, participants will to the outpatient clinic in the morning
      after an overnight fast. Anthropometric data (weight, height, and waist circumference) will
      collected. Body composition will evaluated using bioimpedance (Quantum® apparatus, RJM
      Systems, Michigan) and blood pressure and heart rate (digital monitor, model HEM705CP®,
      Omron) will measured after 30 minutes of rest.

      A catheter with a three-way stopcock will inserted into the arm of the volunteers. Blood
      samples (5mL) will collected after overnight fasting (baseline) and 30, 60, 120, and 240
      minutes after the standardized meal containing sucrose or glucose or fructose overload.
      Participants will remain seated throughout the evaluation period. Participants will receive a
      standardized meal of bread, ham, and margarine plus a sweetened drink (200mL) with similar
      amounts of different carbohydrates (sucrose, glucose, or fructose) in each intervention. The
      meals will provide 25% of the energy requirements, calculated from the resting energy
      expenditure measured by indirect calorimetry (KORR®, MetaCheck) multiplied by the activity
      factor plus 10% referring to the thermal effect of food. The meal will consiste of 15% of
      protein, 30% of fat, and 55% of carbohydrate (30% of complex carbohydrates and 25% of sucrose
      or glucose or fructose).

      Serum levels of glucose, triglycerides, total cholesterol, alanine aminotransferase (ALT) and
      aspartate aminotransferase (AST) will be measured by colorimetric enzymatic test. Serum
      levels of adiponectin, leptin, resistin and TNF will be measured by Enzyme Linked
      ImmuneSorbent Assay (ELISA). Serum levels of IL-2, IL-4, IL-5, IL-6, IL-10, IL-17, IFN-γ and
      eotaxin will be obtained by the Cytometric Bead Array (CBA).
    

Detailed description: 
      This will be a three-arm crossover, randomized, double-blind clinical trial performed at the
      Alpha Institute of Gastroenterology of the Hospital das Clínicas (Belo Horizonte, MG,
      Brazil). All experimental procedures will be conducted according to the guidelines laid down
      in the Declaration of Helsinki. The research protocol was approved by the Institutional
      Review Board of the Universidade Federal de Minas Gerais (CAAE: 21852314.7.0000.5149). All
      volunteers will signed the informed consent prior to the start of the study.

      Sample The sample size of 17 subjects was calculated considering the minimum number of
      participants to detect a difference of at least 11 mg/dL in the levels of triglycerides, 1.2
      ng/mL in adiponectin and 1.9 pg/mL in TNF (pilot study) before and after the intake of a
      single meal containing a fructose overload, 95% confidence interval and statistical power of
      the 80%.

      Study design Each participant will in three visits with washout period between 7 and 21 days.
      At each visit, the participant will pass by a different intervention. Three interventions
      will curr in random order: (i) standardized meal plus sucrose overload; (ii) standardized
      meal plus glucose overload; (iii) standardized meal plus fructose overload. During the
      washout period, the subjects will instructed to maintain their usual eating behavior and
      physical activity. Participants will instructed to avoid consuming caffeine and alcoholic
      beverages and perform intense physical activity within 24 hours before each visit.

      On the day of each intervention, participants will go to the outpatient clinic in the morning
      after an overnight fast. Anthropometric data (weight, height, and waist circumference) will
      collected. Body composition will evaluated using bioimpedance (Quantum® apparatus, RJM
      Systems, Michigan) and blood pressure and heart rate (digital monitor, model HEM705CP®,
      Omron) will measured after 30 minutes of rest.

      A catheter with a three-way stopcock will inserted into the arm of the volunteers. Blood
      samples (5mL) will collected after overnight fasting (baseline) and 30, 60, 120, and 240
      minutes after the standardized meal containing sucrose or glucose or fructose overload.
      Participants remained seated throughout the evaluation period.

      Intervention Participants will receive a standardized meal of bread, ham, and margarine plus
      a sweetened drink (200mL) with similar amounts of different carbohydrates (sucrose, glucose,
      or fructose) in each intervention. The meals will provide 25% of the energy requirements,
      calculated from the resting energy expenditure measured by indirect calorimetry (KORR®,
      MetaCheck) multiplied by the activity factor plus 10% referring to the thermal effect of
      food. The meal will consiste of 15% of protein, 30% of fat, and 55% of carbohydrate (30% of
      complex carbohydrates and 25% of sucrose or glucose or fructose). The sweetened drinks will
      have artificial fruit flavor to avoid the identification of the content by the researchers
      and the volunteers.

      The amount of refined offered carbohydrates to the participants (18 - 25g in a single meal)
      will considered an overload since the World Health Organization and the National Center for
      Chronic Disease Prevention and Health Promotion recommend that the consumption of added
      sugars should not exceed 10% of the total daily calories. For example, in a 2.000 caloric
      diet, no more than 200 calories should come from added sugars (about 12 teaspoons). In
      addition, several authors who investigated short- and long-term (≥ 3 weeks) fructose
      consumption in metabolic and inflammatory biomarkers offered 25% of total energy expenditure
      (GET) in the form of sucrose, glucose, fructose or high fructose corn syrup per day. On the
      other hand, the amount of these carbohydrates offered in the present study is part of the
      eating routine of many individuals (e.g., 200mL of regular soda contains 21 grams of
      sucrose).

      Blood analysis Serum levels of glucose, triglycerides, total cholesterol, alanine
      aminotransferase (ALT) and aspartate aminotransferase (AST) will measured by colorimetric
      enzymatic test (Bioclin®, Belo Horizonte, MG, Brazil). Serum levels of adiponectin, leptin,
      resistin and TNF will measured by Enzyme Linked ImmuneSorbent Assay (ELISA) following the
      manufacturer's instructions (R&D Systems®). Serum levels of IL-2, IL-4, IL-5, IL-6, IL-10,
      IL-17, IFN-γ and eotaxin will obtained by the Cytometric Bead Array (CBA) method (BD
      Bioscience, San Diego, CA, USA) in flow cytometry. Collected blood will diluted in Turk's
      solution. The number of total white blood cell will obtained using a Neubauer chamber (×40
      objective).

      Statistical Analyses All quantitative variables will present as mean ± standard error (SEM).
      The normality of the quantitative variables will tested through the Shapiro-Wilk test.
      Student's t Simple and Mann-Whitney tests will allow comparisons between the initial
      characteristics of those who completed the study and those who did not, and between
      volunteers who will participate in the sub-sample and those who not. To compare the
      anthropometric and clinical characteristics between baseline and the subsequent assessments,
      we will use the ANOVA test for repeated measures 1-factor or Kruskal-Wallis according to the
      normality followed by Bonferroni post-test. Postprandial metabolic and inflammatory markers
      will compared by the Generalized Equation of Estimates (GEE) model to evaluate the effect of
      group allocation (carbohydrate type), time and adjusting for time-group effect (interaction).
      The variables will treated with the connection range function and the correlation matrix will
      the covariance of an unstructured and robust estimator. The Bonferroni post-test will
      identifie the presence of signifcant effects. Significance will set at P < 0.05. The data
      will analysed using the Statistical Package for the Social Sciences (SPSS) software version
      20 and GraphPad Prism version 5.0.
    

Intervention name(s): Group fructose; Group Sucrose; Group Glucose
Intervention type(s): Other; Other; Other
Intervention description(s): Participants will receive a standardized meal of bread, ham, and margarine plus a sweetened drink (200mL) with fructose. The meals will provide 25% of the energy requirements, calculated from the resting energy expenditure measured by indirect calorimetry (KORR®, MetaCheck) multiplied by the activity factor (24) plus 10% referring to the thermal effect of food. The meal will have 15% of protein, 30% of fat, and 55% of carbohydrate (30% of complex carbohydrates and 25% of fructose. The sweetened drinks will have artificial fruit flavor to avoid the identification of the content by the researchers and the volunteers.; Participants will receive a standardized meal of bread, ham, and margarine plus a sweetened drink (200mL) with sucrose. The meals will provide 25% of the energy requirements, calculated from the resting energy expenditure measured by indirect calorimetry (KORR®, MetaCheck) multiplied by the activity factor (24) plus 10% referring to the thermal effect of food. The meal will have 15% of protein, 30% of fat, and 55% of carbohydrate (30% of complex carbohydrates and 25% of sucrose. The sweetened drinks will have artificial fruit flavor to avoid the identification of the content by the researchers and the volunteers.; Participants will receive a standardized meal of bread, ham, and margarine plus a sweetened drink (200mL) with glucose. The meals will provide 25% of the energy requirements, calculated from the resting energy expenditure measured by indirect calorimetry (KORR®, MetaCheck) multiplied by the activity factor (24) plus 10% referring to the thermal effect of food. The meal will have 15% of protein, 30% of fat, and 55% of carbohydrate (30% of complex carbohydrates and 25% of glucose. The sweetened drinks will have artificial fruit flavor to avoid the identification of the content by the researchers and the volunteers.
Other intervention name(s): Placebo Comparator
Allocation: Randomized
Intervention model: Crossover Assignment
Intervention model description: This will be a three-arm crossover, randomized, double-blind clinical trial.
Masking: Triple (Participant, Investigator, Outcomes Assessor)
Primary purpose: Prevention
Keywords: Fructose; Postprandial Period; Inflammation; Metabolism