Dietary Intervention Modifies Gut Microbiota in Type 2 Diabetes.

Study Overview
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Brief Summary
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Aim: To study the effect of a dietary intervention with functional foods on gut microbiota in subjects with type 2 diabetes Materials and methods: Patients with type 2 diabetes were enrolled in a double-blind, parallel-arm, placebo-control study were randomized to receive a dietary portfolio (DP) or placebo (P) treatment for 3 mo. The primary endpoint was the effect of a dietary portfolio based on functional foods on gut microbiota. Secondary endpoints were biochemical parameters, branched chain amino acids, TMAO

Detailed Description
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This study was a single-center, randomized, controlled, double-blind, parallel versus placebo that consisted of six visits. The first visit was a screening evaluation to determine whether subjects meet the inclusion criteria. The selected subjects were invited to a second visit that consisted of a medical history, 2-h oral glucose tolerance test (OGTT), collection of stool samples for DNA isolation and collection of 5 ml blood sample. The participants received the first stage dietary strategy for 15 days. In the third visit and second stage of dietary treatment, subjects were randomized to received the dietary portfolio (DP) or placebo (P) treatment accompanied of the reduced energy diet for 1 mo. In the fourth and fifth visits, with a 1 mo interval, dietary assessment and compliance to the DP or P was evaluated. During each follow-up visit, a 24-h dietary recall was collected, a physical activity questionnaire was filled out and anthropometric and clinical parameters were assessed. In the sixth visit, a 2-h oral glucose tolerance test (OGTT) was performed, and a stool sample for DNA isolation and 5 ml blood were collected. Dietary Intervention In the first stage, the participants consumed a reduced-energy diet tailored to provide a 500-kcal/d deficit as recommended by NIH (8) the with respect to their habitual diet for 15 days. The diet plan consisted in 45-55% carbohydrates, 15-20% protein, 25-35% fat, <7% saturated fat, 200 mg/d cholesterol, 20-35g fiber, 2000-3000 mg/d sodium based on total energy. In the second stage the participants continued to consume the reduced energy diet with the addition of a combination of functional foods (dietary portfolio; DP). The DP provided 200 kcal that were subtracted from the diet. The DP consisted of a mixture of 14g of dehydrated nopal, 4g of chia seed, 30g of soy protein, 4g of inulin,) and 1g of flavoring. The placebo (P) consisted of 28 g of calcium caseinate, 15g of maltodextrin and 1g of flavoring. The kcal, appearance and flavor were similar in DP and P. The DP and P was given in a package in dehydrated form ready to be dissolved in water. The DP was divided into two packages, the first package contained 17.3 g of DP or P given in the breakfast and dissolved in 250 ml and the second package was given at the dinner time (15:00-16:00 h) and contained 34.7g of P and DP dissolved in 300 mL of water. Dietary compliance Dietary compliance was assessed with a 24-h dietary recall and 3-d food record (food log), during each visit that were analyzed by Food Processor Nutrition Analysis Software. The compliance of the consumption of the DP or P was evaluated with the number of empty packages returned at the following visit. Physical activity was assessed using the International physical activity questionnaire (IPAQ).

Official Title
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A Dietary Intervention With Functional Foods Reduce Metabolic Endotoxemia and Attenuates Biochemical Abnormalities in Subjects With Type 2 Diabetes by Modifying the Gut Microbiota.

Conditions
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Type 2 Diabetes Mellitus

Intervention / Treatment
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* Dietary Supplement: dietary portfolio


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: clinical diagnosis of type 2 diabetes Male or female. Adults between 30 and 60 years old. BMI of 25 to 39.9 kg / m². Pharmacological treatment with metformin, a combination of metformin and glibenclamide. Evolution of the type 2 diabetes of 4 ± 3 years. Patients who knew how to read and write. Signature of informed consent. Exclusion Criteria: Diseases that produce secondary obesity. Cardiovascular event. Weight loss> 3 kg in the last 3 months after the evaluation of the criteria. Catabolic diseases such as cancer and acquired immunodeficiency syndrome. Gravidity status. Positive smoking. Treatment with antihypertensive drugs Treatment with other hypoglycemic agents that were not metformin Treatment with statins, fibrates or other drugs to control dyslipidemia, 6 months before the start of the protocol. Any drug or medication that activates intestinal motility Use of laxatives or antispasmodics 4 weeks before the study Treatment with antibiotics 6 months before the study Use of steroids, chemotherapy, immunosuppressant or radiotherapy. Uncontrolled type 2 diabetes, ( HbA1c concentration ≥ 9.9%) Fasting glucose ≥ 220 mg / dL Fasting cholesterol ≥ 240 mg / dL Fasting triglycerides ≥ 350 mg / dL Serum creatinine in women> 1.2 mg / dL in men> 1.3 mg / d

Ages Eligible for Study
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Minimum Age: 30 Years
Maximum Age: 60 Years

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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No

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Treatment
Allocation: Randomized
Intervention Model: Parallel Assignment
Interventional Model Description: In the first stage, the participants with type 2 diabetes consumed a reduced-energy diet tailored to provide a 500-kcal/d deficit as recommended by NIH (8) the with respect to their habitual diet for 15 days. In the second stage the participants continued to consume the reduced energy diet with the addition of a combination of functional foods (dietary portfolio; DP). The DP provided 200 kcal that were subtracted from the diet. The DP consisted of a mi The kcal, appearance and flavor were similar in DP and P. The DP and P was given in a package in dehydrated form ready to be dissolved in water. The DP was divided into two packages, the first package contained 17.3 g of DP or P given in the breakfast and dissolved in 250 ml and the second package was given at the dinner time (15:00-16:00 h) and contained 34.7g of P and DP dissolved in 300 mL of water.
Masking: Double


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: 1. Dietary portfolio (DP)<br>the dietary portfolio was given daily in the breakfast and dinner for 2.5 months | Dietary Supplement: dietary portfolio<br>* The dietary intervention was a combination of functional foods (dehydrated nopal, chia seed, soy protein and inulin) that was provided in dehydrated form in packages of 17.3 g dissolved in 250 ml water for breakfast and 34.7 g in 300 ml water for dinner.<br>|
| Placebo Comparator: 2. placebo (P)<br>the placebo (P) was based was given daily in the breakfast and dinner for 2.5 months | Dietary Supplement: dietary portfolio<br>* The dietary intervention was a combination of functional foods (dehydrated nopal, chia seed, soy protein and inulin) that was provided in dehydrated form in packages of 17.3 g dissolved in 250 ml water for breakfast and 34.7 g in 300 ml water for dinner.<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| intestinal microbiota | Measurement of intestinal microbiota by sequencing using the Illumina platform | Change from baseline gut microbiota at three months after the dietary intervention | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Glucose metabolism profile | serum glucose (mg/dl) | Change from baseline serum glucose at three months after dietary intervention | 
| Glucose metabolism profile | serum insulin (µUI/ml) | Change from baseline serum insulin at three months after dietary intervention | 
| Glucose metabolism profile | plasma glycated hemoglobin (HbA1c) (%) | Change from baseline plasma HbA1c at three months after dietary intervention | 
| Lipid metabolism profile | serum triglycerides (mg/dl) | Change from baseline serum triglycerides at three months after dietary intervention | 
| Lipid metabolism profile | serum total cholesterol (mg/dl) | Change from baseline serum total cholesterol at three months after dietary intervention | 
| Lipid metabolism profile | serum LDL cholesterol (mg/dl) | Change from baseline serum LDL cholesterol at three months after dietary intervention | 
| Lipid metabolism profile | serum HDL cholesterol (mg/dl) | Change from baseline serum HDL cholesterol at three months after dietary intervention | 
| Lipid metabolism profile | plasma free fatty acids (FFA) (mmol/L) | Change from baseline plasma free fatty acids at three months after dietary intervention | 
| metabolomic profile | plasma betaine (µmol/L) | Change from baseline plasma betaine at three months after dietary intervention | 
| metabolomic profile | plasma choline (µmol/L) | Change from baseline plasma choline at three months after dietary intervention | 
| metabolomic profile | plasma trimethylamine oxide (TMAO) (µmol/L) | Change from baseline plasma TMAO at three months after dietary intervention | 
| metabolomic profile | plasma branched chain amino acids (BCAA) (µmol/L) | Change from baseline plasma BCAA at three months after dietary intervention | 
| inflammatory profile | plasma lipopolysaccharide (LPS) (ng/ml) | Change from baseline plasma LPS at three months after dietary intervention | 
| inflammatory profile | serum C reactive protein (CRP) (mg/dl) | Change from baseline serum CRP at three months after dietary intervention | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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gut microbiota, functional foods, branched chain amino acids, TMAO, type 2 diabetes