Effects of Intermittent Fasting in Metabolic Dysfunction Associated Fatty Liver Disease
Phase: NA
Conditions: Intermittent Fasting, Metabolic Dysfunction-Associated Fatty Liver Disease
Sponsor: Istanbul Bilgi University

Official Title of the study: Effects of Intermittent Fasting in Metabolic Dysfunction 
Associated Fatty Liver Disease 
NCT number: NCT06664684 
Date of the document: 06.05.2022 
Study Protocol 
Patients  
This study was approved by the Ethics Committee of Marmara University School of Medicine 
(No: 09.2022.707) Informed consent was obtained from each patient included in the study. 
This is a prospective non-drug intervention study conducted by the same gastroenterologist 
on patients diagnosed with MAFLD. Individuals with fatty liver disease diagnosed by 
vibration-controlled transient elastography are diagnosed with MAFLD if at least one of the 
following three conditions is present: overweight/obesity, type 2 diabetes mellitus (T2DM), or 
underweight or normal weight with at least two metabolic risk abnormalities [waist 
circumference, blood pressure, triglycerides, high-density lipoprotein cholesterol (HDL-C), 
prediabetes, insulin resistance, high-sensitivity C reactive protein (CRP)]. All of our patients 
were diagnosed with MAFLD because their BMI was above 25 kg/m2 in addition to the 
detection of fatty liver.  
Exclusion criteria were patients with an average daily alcohol consumption of >20 g for 
females and >30 g for males; pregnant or lactating women; patients with ischemic heart 
disease or heart failure, chronic inflammatory diseases, chronic viral infections, cancer, 
moderate-to-severe kidney disease, uncontrolled hypertension, and eating disorders; those 
with a history of bariatric surgery; and those on insulin due to increased risk of hypoglycemia.  
Design  
Patients who voluntarily agreed to participate in the study were randomly assigned into two 
groups. Patients enrolled in the study were referred for an initial consultation with a dietitian 
following their diagnosis by a gastroenterologist. The energy-restricted diet group followed an 
energy-restricted diet for 8 weeks, while the energy + time-restricted diet group followed the 
same diet and a 16:8 eating pattern and restricted food intake to an 8 h window each day. The 
effectiveness of the intervention was assessed using anthropometric and biochemical 
measurements, and Fibroscan investigations were performed before the intervention (T0) 
and after the intervention (T8).  
Measurements  
Blood samples were collected from patients after 12 h of fasting. The samples were 
centrifuged for 15 min and stored at −80 ◦C after serum separation until analysis. Serum 
FGF-21, Beclin-1, and ATG-5 were analyzed using enzyme-linked  
 

 
immunosorbent assay (ELISA) kits, following the manufacturer’s protocols (Human FGF-21 
ELISA, Biovendor, Brno, Czech Republic; Human BECN1 [Beclin-1] ELISA Kit, ElabScience, 
Houston, TX, USA; Human Autophagy protein 5 [ATG5] ELISA Kit, MyBioSource, Inc., San 
Diego, USA) and in duplicate.  
- The extent of hepatic steatosis and fibrosis was determined using transient elastography on 
a Fibroscan 502 TouchR device(Echosens SA, Paris, France). All Fibroscan measurements 
were performed following the manufacturer’s instructions as specified previously.  
- Patients’ heights were measured using a stadiometer (Seca 769), barefoot, feet side by side, 
and head in the Frankfort plane. Body composition was determined using the bioelectrical 
impedance method (Tanita MC 780 P).