High-intensity Interval Training Versus Moderate Intensity Continuous Training on Gut Microbiota in Non-alcoholic Fatty Liver

non- Alcoholic Fatty Liver disease affects approximately 25% of the global population, with its prevalence increasing due to lifestyle changes such as poor diet and sedentary behavior. non- Alcoholic Fatty Liver disease is often associated with metabolic disorders, including insulin resistance, obesity, and type 2 diabetes. Emerging evidence suggests that alterations in gut microbiota may contribute to the pathophysiology of non- Alcoholic Fatty Liver disease, influencing liver function, inflammation, and metabolic regulation. However, there is limited research exploring the effects of interventions such as exercise and dietary changes on gut microbiota composition in non- Alcoholic Fatty Liver disease patients The findings of this study could have important clinical implications, particularly for developing tailored lifestyle interventions for non- Alcoholic Fatty Liver disease patients. As gut microbiota dysbiosis is increasingly recognized as a key player in the development and progression of non- Alcoholic Fatty Liver disease, interventions targeting microbiota balance could offer novel therapeutic approaches for managing this prevalent and potentially severe condition

• High intensity interval training (Other): Patients will engage in high-intensity interval training using heart rate reserve (HRR) to determine exercise intensity. The program includes three sessions per week for 12 weeks, featuring three 3-minute high-intensity intervals at 70-90% HRR, with 2 minutes of active recovery in between. Each 30-minute session will be treadmill-based under supervision, incorporating warm-up and cool-down periods. Participants will also follow a Mediterranean diet.
• moderate intensity continuous training (Other): The program will be in form of: * Frequency: 3 sessions/week for 12 weeks. * Intensity: Moderate intensity continues training at 40- 60% HRR. * Duration: the session will last 30 minutes per session. Patients will be instructed to follow Mediterranean Diet.
• Mediterranean diet (Other): the composition of the Mediterranean diet, emphasizing that vegetables and fruits make up the largest portion at 35%, followed by whole grains (20%) and fish and seafood (15%). Legumes and nuts account for 10%, while olive oil and dairy products (cheese and yogurt) each contribute 7%. Smaller portions include poultry and eggs (4%) and red meat and sweets (2%), reflecting the diet's focus on plant-based foods, healthy fats, whole grains, and moderate consumption of animal products, with minimal intake of red meat and sugary foods.

Primary outcomes

• assessment of short chain free fatty acids butyrate and acetate [at baseline and after 12 weeks]: For short chain free fatty acids analyses, fecal sample will be separated. Applying four analytes, including total Short Chain Fatty Acids, acetate, propionate, and butyrate, will be targeted. Final concentrations will be calculated based on internal standards and are expressed as micromoles per gram of wet feces (μmol/g). Techniques and predictive approaches will be used across. gas chromatography will be used to quantify Short Chain Fatty Acids from fecal samples
• assessment of lipid profile [at baseline and after 12 weeks]: Fasting blood samples were collected after an overnight fast, and serum lipid levels were analyzed using standard enzymatic methods with an automated biochemical analyzer. The lipid profile included triglycerides, total cholesterol, low-density lipoprotein cholesterol, and high-density lipoprotein cholesterol to assess lipid metabolism and cardiovascular risk.

Secondary outcomes

• assessment of quality of life [at baseline and after 12 weeks]: The short form-12 was scored using the standard scoring algorithm, which converts participants' responses into two norm-based summary measures: the Physical Component Summary and the Mental Component Summary. Each score ranges from 0 to 100, with higher scores indicating better health-related quality of life. The scores were calculated according to the official short form -12 scoring guidelines.

Sex: All, ages: 40 Years to 60 Years

Inclusion Criteria:

1. Patients from both genders aged 40 - 60 years old diagnosed with Non-alcoholic Fatty Liver disease by ultrasonography.
2. Body mass index will range from 30 to 34.9 kg/m².
3. Patients with central obesity (waist circumference 90 cm for men and 85 cm for women).
4. Patients who have taken no regular exercise in the last year (<3 times/week and 30 min/time.
5. No antibiotic or probiotic use for at least last 3 months.
6. Medical drugs such as metformin, sulfonylureas, statins, and blood pressure medication were eligible, as well as stable.

Exclusion Criteria:

1. History of liver cirrhosis or other chronic liver diseases.
2. Use of medications affecting gut microbiota (e.g., laxatives, metformin, proton pump inhibitors).
3. Recent gastrointestinal infections or surgeries.
4. Diabetes Mellitus type 1 or uncontrolled type 2.
5. Cardiovascular, respiratory, or musculoskeletal disorders limiting exercise capacity.
6. Active smokers.
7. Pregnancy or breastfeeding.
8. Uncontrolled hypertension.
9. Communication disorders.
10. Bleeding disorders.