Mitophagy and Mitochondrial DNA Dynamics During Ramadan Dry Fasting vs 16:8 Time-Restricted Feeding

Official title

Mitophagic Flux and Mitochondrial DNA Dynamics During Circadian-Aligned Ramadan Dry Fasting Versus 16:8 Time-Restricted Feeding: A Four-Phase Translational Research Programme

Overview

This study investigates how different types of fasting affect cell health, specifically focusing on how mitochondria (the energy-producing parts of cells) are cleared and renewed. While standard intermittent fasting allows water intake, Ramadan dry fasting involves total restriction of both food and fluids from dawn to sunset. Researchers want to see if the combined effects of fluid restriction and natural daily body rhythms during Ramadan trigger a stronger cellular cleanup process (called mitophagy) compared to standard 16:8 water-permitted fasting. The study will look at how these fasting habits change blood markers related to mitochondrial DNA and metabolic health across a 30-day period. This research will help determine if dry fasting offers distinct biological benefits for cellular renewal.

Detailed description

This is a prospective, single-centre, parallel-group, longitudinal observational cohort study conducted across Ramadan 2027. The study aims to evaluate the distinct molecular and metabolic trajectories of intermittent dry fasting compared to standard fluid-permitted time-restricted feeding (TRF).

Participants will be allocated to one of two parallel groups based on their self-selected, established fasting practices:

* Group A (Ramadan Dry Fasting): Observing complete fluid and nutrient deprivation from dawn (Fajr) to sunset (Maghrib) for 30 consecutive days. * Group B (16:8 TRF): Practicing a 16-hour daily fasting window with water/fluids permitted, observed concurrently during the same calendar period.

Participants in both groups will be pairwise-matched based on age and BMI at screening. Blood samples will be collected from all participants at five distinct milestones:

* T0: Pre-Ramadan baseline (Day -3) * T1: Acute induction phase (Day 7) * T2: Mitochondrial turnover phase (Day 14) * T3: Saturation plateau / primary endpoint (Day 30) * T4: Post-Ramadan recovery phase (Day 44)

Peripheral blood mononuclear cells (PBMCs) and serum will be isolated from the collected blood. Laboratory analyses will measure mitochondrial DNA (mtDNA) copy number changes via qPCR, alongside a quantitative gene expression panel evaluating key regulators of mitophagy (such as PINK1 and Parkin), mitochondrial biogenesis, and general autophagy. Secondary evaluations include routine serum metabolic indicators, inflammatory markers, and dietary monitoring to evaluate the physiological pathways predicted by computational systems pharmacology.

Interventions

• Behavioral Ramadan Intermittent Dry Fasting (Sawm)
  Total fluid and nutrient restriction enforced daily from dawn (Fajr) to sunset (Maghrib) for 30 consecutive days. Food and fluid consumption are restricted to the nocturnal period
• Behavioral 16:8 Time-Restricted Feeding (TRF)
  A 16-hour daily fasting window where calorie-containing foods and beverages are completely restricted. Fluid intake (such as water or uncaloried drinks) is permitted throughout the fasting period

Primary outcome measures

• Between-arm difference in PINK1 and Parkin mitophagy gene expression [Time frame: Change from baseline (Day -3) to Day 30 of the fasting period]
• Between-arm difference in mitochondrial DNA copy number (mtCN) [Time frame: Change from baseline (Day -3) to Day 30 of the fasting period]

Eligibility criteria

Inclusion criteria

• Common Criteria (Both Arms):
• Age 18-50 years
• BMI 18.5-30.0 kg/m²\[cite: 1\]
• Non-smoker for at least 12 months\[cite: 1\]
• No chronic disease requiring regular prescription medication\[cite: 1\]
• Stable sleep-wake schedule (self-reported sleep midpoint deviation less than 90 minutes)\[cite: 1\]
• Willing and able to attend all five study visits at the specified times\[cite: 1\]

Arm A Specific (Ramadan Dry Fasting):

\- Documented religious commitment and intention to complete all 30 days of Ramadan 2027 fasting\[cite: 1\]

Arm B Specific (16:8 TRF Control):

• Confirmed practice of 16:8 TRF (fasting window 16 hours or more per day) throughout the study period\[cite: 1\]
• Not observing Ramadan dry fasting during the study period\[cite: 1\]

Exclusion criteria

• Established type 1 or type 2 diabetes (HbA1c 6.5% or higher or pharmacological antidiabetic therapy)\[cite: 1\]
• Severe gastrointestinal, cardiovascular, hepatic, or renal disease (eGFR less than 60 mL/min/1.73m²)\[cite: 1\]
• Active cancer or chemotherapy within five years\[cite: 1\]
• Use of metformin, statins, or exogenous antioxidant supplements (vitamins C, E, NAC, CoQ10) within 30 days of screening\[cite: 1\]
• Pregnancy, lactation, or planned conception during the study period\[cite: 1\]
• Participation in another clinical study within three months\[cite: 1\]
• Inability to provide written informed consent in Arabic or English\[cite: 1\]

Criteria are shown verbatim from the registry (in English). Final eligibility is always assessed by the study center.

Healthy volunteers: Yes

Study design

Observational model

Cohort

Study locations

Cyprus · 1 center

• Near East University — Nicosia

Publications

• Sarkar B, Hajra A. Hydro-phosphorothiolation of Styrene and Cyclopropane with S-Hydrogen Phosphorothioates under Ambient Conditions. Org Lett. 2024 Jun 21;26(24):5141-5145. doi: 10.1021/acs.orglett.4c01586. Epub 2024 Jun 7. PMID 38848455
• Dillard J, Meng X, Nelin L, Liu Y, Chen B. Nitric oxide activates AMPK by modulating PDE3A in human pulmonary artery smooth muscle cells. Physiol Rep. 2020 Sep;8(17):e14559. doi: 10.14814/phy2.14559. PMID 32914566

Identifiers

NCT: NCT07638696 · Bahcesehir Cyprus University

Primary sources (government registries)

• ClinicalTrials.gov (US NIH)

View this study on ClinicalTrials.gov ↗