Efficacy of a Nutritional Education Strategy and Physical Exercise on the Gut Microbiota in Type 2 Diabetics

NCT05261373 | Unknown

• 1 other identifier: The EDUGUTION project
• Study Type: interventional
• Target: 120
• Locations: 1 country
• Sites: 1

Brief Summary

We hypothesize that the combination of a nutritional education intervention with a HIIT-based physical exercise program improve muscle metabolism through positive modifications of gut microbiota in people with T2DM, leading to better glycaemia/insulinaemia levels, reduction of body fat mass and improving quality of life. The project is a randomized controlled clinical trial in 120 participants with T2DM and obesity, which aims to determine the efficacy of a nutritional education program and the role of physical exercise type on health related variables. The participants will be of both sexes with age between 40 and 55 years, belonging to the Province of Cádiz. The design has two 12-week interventions; the main factor has 2 levels: participants who receive the nutritional education (EDU) and controls (CG); the second factor has 3 levels: high-intensity interval training (HIIT), moderate intensity continuous training (MICT), and controls (INACT). Therefore, participants will be randomized into 6 groups (n=20), adjusted by gender (≈50% in each group): EDU+HIIT, EDU+MICT, EDU+INACT, CG+HIIT, CG+MICT, CG+INACT. The outcome variables, which will be measured before and after the intervention, will include: dietary intake assessment, physical activity assessment, quality of life, faecal samples, blood samples, blood pressure, appetite assessment, muscle biopsy samples, body composition and fluids, basal metabolism, maximal fat oxidation test and cardiorespiratory fitness.

Conditions

Diabetes Mellitus, Type 2; Cardiovascular Diseases; Fat Burn; Metabolism Disorder

Keywords

Gut microbiota; High intensity interval training; Insulin resistance; Diet; Fat metabolism

Outcome Measures

Primary Outcomes (7)

• Assessed gut microbiota population

  For the DNA extraction, fecal samples will be homogenized in a Stomacher-400 mixer. The DNA will be extracted using a mini QIAamp DNA stool kit (QIAGEN, Barcelona, Spain) as indicated by the manufacturer. Quantification is performed with a NanoDrop ND-1000 spectrophotometer (Thermo Fisher Scientific, DE, USA). For sequencing analysis, DNA extracted is amplified by Polymerase Chain Reaction (PCR). All PCRs will be performed in 25 μl of reaction volumes containing 12.5 μl of the KAPA HiFi Hotstart 2X prepared mixture (KAPA Biosystems, Woburn, MA), 5 μl of each direct and reverse primer (1 μM) and 2.5 μl of extracted DNA (10 ng) under standardized cycling conditions. PCR cleaning will be performed with beads and PCR products combined with AMPure XP beads (Beckman Coulter, USA). Determine gut microbiota composition and characterization, for classification of phylum, genus, and species of bacteria. Specifically the Firmicutes/Bacteroidetes ratio.

  12 weeks

• Assessed changes in Insulin Resistance

  They will be determined using the ELISA technique for the concentration of insulin. The Homeostatic Model Assessment for Insulin Resistance insulin sensitivity index (HOMA-IR) will be calculated.

  12 weeks

• Muscle biopsy samples: mRNA expression by quantitative real-time PCR (Transcriptomic)

  The mRNA levels of Glucose Transporter GLUT4, IRS1, PI 3-Kinase, PGC1-alpha, SIRT1, Mitofusin 1, Mitofusin 2, Complex I, III, IV, V of mitochondria (Nd1, Cyb, Co1, Atp6), CPT1B, and OPA1 will be analysed, for that, total RNA will be isolated from tissues (frozen muscle biopsies) an Illustra MiniRNA Spin kit (GE Healthcare). Then, cDNA will be obtained using a PrimeScript™ RT Master Mix (Perfect Real Time) (Takara), following the manufacturer's instructions. Relative quantification of mRNA will be performed using a Corbett RotorGene by using 6.5 ng of cDNA, forward and reverse primers at 100 nM each, and a SYBR Green PCR Master Mix Reagent kit (Life Technologies) in 10 μL of reaction. The mRNA levels will be normalized to those of housekeeping genes (b-actin, a-tubulin) and expressed as fold change.

  12 weeks

• Muscle biopsy samples: Western blot analysis (Proteomic)

  Frozen muscle tissue will be homogenized in protein extraction buffer (RIPA buffer with phosphatase inhibitors and protease inhibitors). Protein concentration will be determined using a BCA protein assay kit (Thermoscientific). Then, samples will be separated on SDS-PAGE gels, and then transferred onto PVDF membranes (Millipore). The following primary antibodies will be used Rabbit polyclonal to PGC1 alpha, Rabbit monoclonal \[EPR19274\] to DRP1, Rabbit monoclonal \[E104\] to SIRT1, Rabbit monoclonal \[EP2109Y\] to AKT1 (phospho S473), Rabbit polyclonal to TIM44, Rabbit polyclonal to Glucose Transporter GLUT4, Rabbit polyclonal to IRS1, Rabbit polyclonal to AKT1, Rabbit monoclonal \[EPR5683\] to AMPK alpha 2 (phospho T172), Rabbit polyclonal to CPT1B, Total OXPHOS Human WB Antibody Cocktail, Rabbit polyclonal to Mitofusin 1, Rabbit polyclonal to OPA1, Mouse monoclonal \[12C4F12\] to MTCO2, Rabbit polyclonal to IRS1 (phospho S312), Rabbit polyclonal to AMPK alpha 2, Mouse monoclonal \[6A

  12 weeks

• Muscle biopsy samples: Citrate synthase (CS) activity assay (Mitochondrial content marker)

  Therefore, skeletal muscle (5 mg) will be homogenized in 100 μl cold CelLytic MT (Sigma) at pH 7.4 and protease inhibitor cocktail. The homogenized sample will be centrifuged at 12,000xg for 10 min and the supernatant containing the protein collected. After, 8 μg of protein, as determined by BCA protein assay. In addition to protein, the reaction mixture contained 1X assay buffer, 300 μM acetyl CoA and 100 μM 5,5'-Dithiobis-(2-nitrobenzoic acid). The reaction starts by adding 500 μM Oxaloacetate. CS activity is measured by continuous spectrophotometric rate determination at 412 nm, according to manufacturer instructions. Each sample will be run in triplicate.

  12 weeks

• Assessed changes from Appetite assessment

  Since appetite feeling modulates nutritional behavior, the evaluation of appetite among the intervention groups can improve the quality of the study from a comprehensive perspective. After a period of 8-10 hours of fasting, an analog visual scale (AVS) will be completed in order to ensure the appetite felt by the participant in the morning. The AVS runs from 1 to 10, being the lowest value no appetite at all, and the maximum full appetite.

  12 weeks

• Assessed changes from Body composition: Fatmass and Fat-free mass.

  Body composition will be estimated using a multifrequency bioimpedance of 8 electrodes previously validated (TANITAMC780MA). The calculation of impedance can estimate the fat mass and fat-free mass in kilograms. The patients will wear light clothing and will assume a posture in accordance with the manufacturers' instructions. Other previous considerations will be followed 24 hours before the measure: (i) to refrain from vigorous exercise, (ii) to take alcoholic drinks, (iii) to take energy drinks, and (iv) to be in a fasting state for at least 8 hours. Hydration status will be controlled through a urine color scale from clear to dark during the 7 days before assessment for adjusting variables.

  12 weeks

Secondary Outcomes (9)

• Assessed changes from dietary intake: Frequency of consumption

  12 weeks

• Assessed changes from dietary intakes: 24 hours dietary recalls

  12 weeks

• Assessed changes from accelerometry: Physical activity time

  12 weeks

• Assessed changes from physical activity

  12 weeks

• Assessed changes from resting fat oxidation

  12 weeks

Study Arms (6)

Control (CG+INACT)

NO INTERVENTION

Participants who do not receive neither nutritional education nor exercise program. They will be instructed to maintain their normal life habits with respect to physical activity and diet.

Moderate-intensity continuous training (CG+MICT)

ACTIVE COMPARATOR

Participants who do not receive nutritional education but are enrolled in a moderate-intensity continuous training exercise program.

Other: The Moderate-Intensity Continuous Training (MICT)

High-intensity interval training (CG+HIIT)

ACTIVE COMPARATOR

Participants who do not receive nutritional education but are enrolled in a high-intensity interval training exercise program.

Other: The High-Intensity Interval Training (HIIT)

Nutritional Education (EDU+INACT)

ACTIVE COMPARATOR

Participants who receive nutritional education but not an exercise program.

Other: Behavioral: Nutritional education program

Nutritional Education Moderate-intensity continuous training (EDU+MICT)

EXPERIMENTAL

Participants who receive nutritional education and are enrolled in a moderate-intensity continuous training exercise program.

Other: The Moderate-Intensity Continuous Training (MICT); Other: Behavioral: Nutritional education program

Nutritional Education High-intensity interval training (EDU+HIIT)

EXPERIMENTAL

Participants who receive nutritional education and are enrolled in a high-intensity interval training exercise program.

Other: The High-Intensity Interval Training (HIIT); Other: Behavioral: Nutritional education program

Interventions

The Moderate-Intensity Continuous Training (MICT) OTHER

3 sessions per week in a cycle ergometer, with 1-2 days off between sessions, during a total of 12 weeks under the supervision of a personal trainer. MICT program will consist of sessions of approximately ˜45 min of moderate aerobic exercise (at the intensity at which the maximal fat oxidation was achieved in the laboratory test) in cycle ergometer.

Moderate-intensity continuous training (CG+MICT); Nutritional Education Moderate-intensity continuous training (EDU+MICT)

The High-Intensity Interval Training (HIIT) OTHER

3 sessions per week in a cycle ergometer, with 1-2 days off between sessions, during a total of 12 weeks under the supervision of a personal trainer. HIIT program will consist of 3-5 series of 1 min duration at 90-130% of its maximum power (determined in the first training session), with 90 seconds of rest between sets (estimated total time of the session: 10 minutes). Depending on the number of training session, the load and the series will increase up to 40% more than the maximum load.

High-intensity interval training (CG+HIIT); Nutritional Education High-intensity interval training (EDU+HIIT)

Behavioral: Nutritional education program OTHER

The education intervention will consist of individual nutritional counselling. The nutritional education program will be conducted every 2 weeks for 12 consecutive weeks, with 20-min counselling sessions by an experienced nutritionist. Participants will be provided with an introduction (in an easy-to-understand manner) regarding the association between T2DM, gut microbiome and dietary habits. Firstly, the diet of the patient should be analysed, to determine which aspects can be improved, such as, total calories intake, amount and types of carbohydrates (highlighting the relevance of fibre), etc. Moreover, some suggestions about the combination of foods and culinary technical in order to manage the glycaemic index of foods will be provided. Finally, those pattern of the Mediterranean diet and some qualitative aspects and aids (as the size of the plat or avoid having the food platter on the table) will be taught the patients.

Nutritional Education (EDU+INACT); Nutritional Education High-intensity interval training (EDU+HIIT); Nutritional Education Moderate-intensity continuous training (EDU+MICT)

Eligibility Criteria

• Age: 40 Years - 55 Years
• Sex: all
• Healthy Volunteers: No
• Age Groups: Adult (18-64)

You may qualify if:

• Non smoking
• Non-alcoholic (\<3 standard drinks per day)
• Body mass index \>25 kg/m maintaining the habitual dietary patterns without a body mass reduction higher than 2% during the last 6 months
• Not being insulin dependent
• Absence of injury, disease or disability or other known medical condition which could affect the ability to successfully participate in physical exercise tests
• Absence of cardiovascular disease (angina, peripheral or cerebro-vascular disease, etc.).
• Absence of neurologic and psychiatric diseases.
• Absence of respiratory diseases (pulmonary hypertension, Chronic obstructive pulmonary disease, etc.).
• Absence of other metabolic diseases (hyper/hypo parathyroidism, hyper/hypothyroidism, Cushing's disease, Type 1 diabetes, etc.)
• Absence of active inflammatory bowel disease
• Absence of kidney disease
• Absence of tumours
• Absence of coagulation dysfunction
• Not under treatment with medications k known to affect glucose metabolism, recent steroid treatment (within 6 months), or hormone replacement therapy
• Be able to understand a communication in Spanish or English.

You may not qualify if:

• They do not attend more than 2 or 4 consecutive sessions of nutritional counselling or physical training respectively.
• The lose more than 4 or 6 sessions in total of nutritional counselling or physical training respectively.

Sponsors & Collaborators

• University of Cadiz: lead
• Ministerio de Ciencia e Innovación, Spain: collaborator

Study Sites (1)

Facultad de Ciencias de la Educación

Puerto Real, Cadiz, 11519, Spain

RECRUITING

MeSH Terms

Conditions

Diabetes Mellitus, Type 2; Cardiovascular Diseases; Metabolic Diseases; Insulin Resistance

Condition Hierarchy (Ancestors)

Diabetes Mellitus; Glucose Metabolism Disorders; Nutritional and Metabolic Diseases; Endocrine System Diseases; Hyperinsulinism

Study Officials

• Jesús Gustavo Ponce González, PhD

  University of Cádiz

  PRINCIPAL INVESTIGATOR

• Cristina Casals, PhD

  University of Cádiz

  PRINCIPAL INVESTIGATOR

Central Study Contacts

Alberto Marín Galindo, MsC

CONTACT

685847806; alberto.marin@uca.es

Jesús Gustavo Ponce González, PhD

CONTACT

658874134; jesusgustavo.ponce@uca.es

Study Design

• Study Type: interventional
• Phase: not applicable
• Allocation: RANDOMIZED
• Masking: SINGLE
• Who Masked: OUTCOMES ASSESSOR
• Purpose: TREATMENT
• Intervention Model: PARALLEL
• Sponsor Type: OTHER
• Responsible Party: SPONSOR

Model Details: The project is a randomized controlled clinical trial with T2DM and obesity

Study Record Dates

• First Submitted: December 17, 2021
• First Posted: March 2, 2022
• Study Start: March 10, 2022
• Primary Completion: September 1, 2023
• Study Completion: August 31, 2024
• Last Updated: November 3, 2022

Record last verified: 2022-02

Data Sharing

• IPD Sharing: Will not share

Locations

ES (1)