Nonalcoholic Fatty Liver DiseaseMASLDNCT07740109
The Effects of 5-methyltetrahydrofolate Supplementation in Patients With Metabolic Dysfunction-Associated Steatotic Liver Disease
A study testing 5-methyltetrahydrofolate (5-MTHF) in people with Nonalcoholic Fatty Liver Disease and Nonalcoholic Fatty Liver Disease (NAFLD), run by Tabriz University of Medical Sciences at 1 site in Iran.
Trial summary
To determine the effect of MTHF supplementation on serum folate and homocysteine level, metabolic, nutritional status, liver function, and PPARα and TNFα gene expression in patients with MASLD Read more about this study
- Who is it for?
- Ages 18 to 50 years
- Conditions the study is for
- Nonalcoholic Fatty Liver Disease, Nonalcoholic Fatty Liver Disease (NAFLD), MASLD - Metabolic Dysfunction-Associated Steatotic Liver Disease
- What is being tested?
- 5-methyltetrahydrofolate (5-MTHF)
- Type of treatment
- Pill or oral medicine
- Could I receive a placebo?
- Yes, some participants may receive a placebo
- How many people
- 44 participants
- Where
- 1 siteIran
- Study dates
- 30 Sep 2026to 30 Aug 2027
- Phase
- Interventional
- Are there results?
- Not yet: the study is under way
Key requirements
Who can take part, as listed by the sponsor. Only the study team can confirm whether you are eligible, so take this list to your doctor.
- Ages 18 to 50 years
- All sexes
- No healthy volunteers
- Adult
Who may be able to join
- Adult men or women (18-50 years)
- Diagnosis of MASLD (grade 1 or 2 of steatosis confirmed by ultrasound)
- Body mass index (BMI) = 25-34.9 kg/m²
- Providing written informed consent
Who cannot join
- Pregnancy, lactation, or plans to get pregnant during the next three months.
- Liver disease (viral hepatitis, autoimmune liver disease, cirrhosis, drug-induced hepatotoxicity, or alcoholic fatty liver disease), heart or renal failure, kidney stones, any neoplasia, inflammatory disease, hypothyroidism, hypercortisolism, or hypertension
- Taking drugs affecting glucose or lipid metabolism, folate supplements, anti-obesity medications, weight-loss diets, or dietary supplements
- Lifestyle factors known to impact folate status (current smoking, alcohol intake, recreational drug use)
- Pre-existing conditions affecting folate status (malabsorptive or inflammatory bowel diseases, active celiac disease, gastric bypass surgery, atrophic gastritis, epilepsy, advanced liver disease, kidney dialysis, type 1 or 2 diabetes mellitus, or sickle cell trait/anemia)
- Medications that interfere with B-vitamin metabolism (chloramphenicol, methotrexate, metformin, sulfasalazine, phenobarbital, phenytoin, primidone, triamterene, barbiturates)
Trial locations and contacts
1 site in 1 country. Contact the study team or a site near you to ask about taking part.
Bahram Pourghassem Gargari, PhD. Professor at TUMS contact
Iran1 site
Liver Clinic of Valiasr Hospital (Tabriz, Iran)
Tabriz
Fatemeh Tamjid, MSc, PhD student, +989144755462, tamjid.f@tbzmed.ac.ir
Bahram Pourghassem Gargari, PhD. Professor at TUMS, +989143165247, pourghassemb@tbzmed.ac.ir, bahrampg@yahoo.com
About this study
Official title: The Effect of 5-methyltetrahydrofolate Supplementation on Serum Folate and Homocysteine Level and PPARα and TNFα Gene Expression in Patients With Metabolic Dysfunction-Associated Steatotic Liver Disease: a Double-blind, Parallel Randomized Controlled Trial Study
To determine the effect of MTHF supplementation on serum folate and homocysteine level, metabolic, nutritional status, liver function, and PPARα and TNFα gene expression in patients with MASLD
Read the detailed description
Metabolic dysfunction-associated steatotic liver disease (MASLD), formerly termed non-alcoholic fatty liver disease (NAFLD), is diagnosed via liver biopsy or imaging when steatosis is present in the absence of alcohol intake or other hepatic disorders. As the liver manifestation of metabolic syndrome, it commonly coexists with obesity, diabetes, dyslipidemia, hypertension, and related conditions. Global prevalence of MASLD continues to rise.
Evidence from an earlier systematic review and meta-analysis indicated that MASLD patients had significantly lower serum folate and higher homocysteine concentrations. Folate is an essential water-soluble B vitamin that occurs in multiple chemically related forms. Food folates are mainly reduced and polyglutamated, with 5-MTHF predominating in both the diet and systemic circulation. 5-MTHF does not require reduction by DHFR and can enter the bloodstream directly for use. Reduced folates act as methyl donors in one carbon metabolism, supporting cellular proliferation, homocysteine re-methylation to methionine, nucleic acid synthesis and methylation of DNA, RNA, proteins and phospholipids.
Experimental studies have demonstrated that diet-induced hyperhomocysteinemia promotes hepatic steatosis and liver injury and folate as a key regulator of homocysteine concentration, may exert hepatoprotective effects. Evidence suggests that folate may improve hepatic lipid metabolism by activating peroxisome proliferator-activated receptor alpha (PPARα) signaling and modulate the immune response and reduce inflammatory mediators. Nevertheless, no evidence on the effects of folate on PPARα and TNFα gene expression in MASLD patients exist. Moreover, PPARα gene expression is dysregulated in MASLD and related metabolic conditions; PPARα is highly expressed in the liver, skeletal muscle and brown adipose tissue, stimulates β-oxidation and suppresses fatty-acid synthesis. Although the effect of 5-MTHF supplementation on gene expression of PPARα and TNFα in MASLD patients has not been examined, evidence showed that folate can modulate PPARα and TNFα. As folate has been shown to affect lipid metabolism and inflammation, we hypothesized that 5-MTHF supplementation might regulate PPARα and TNFα expression in MASLD patients. This randomized, double-blind, placebo-controlled clinical trial will therefore be undertaken to determine the effects of 5-MTHF supplementation on serum levels of folate and homocysteine, and gene expression of PPARα and TNFα in MASLD patients.
- Conditions
- Nonalcoholic Fatty Liver Disease, Nonalcoholic Fatty Liver Disease (NAFLD), MASLD - Metabolic Dysfunction-Associated Steatotic Liver Disease, MASLD, NAFLD, Metabolic Dysfunction-Associated Steatotic Liver Disease
- Being tested
- 5-methyltetrahydrofolate (5-MTHF), placebo group
- Keywords
- 5-methyltetrahydrofolate, homocysteine, MASLD, NAFLD, Nonalcoholic Fatty Liver Disease, Metabolic Dysfunction-Associated Steatotic Liver Disease
- Medical subject headings
- Non-alcoholic Fatty Liver Disease
What participants receive
What participants receive. In a randomised study each person is assigned to one group by chance.
5-methyltetrahydrofolate (5-MTHF)Dietary supplement
Patients in this group will receive 5-methyltetrahydrofolate tablets (800 mcg) once a day for 90 days. Tablets will be manufactured by Ashbal Chemi pharmaceutical company (Qfol, Ashbal Chemi Co., Tehran, Iran).
placebo groupDietary supplement
Patients in this group will receive placebo for 90 days. The placebo is corn starch/ cellulose and will be consumed once a day. Placebo tablets will be manufactured by Ashbal Chemi Co. (Tehran, Iran).
| Study group | Role | What they receive |
|---|---|---|
| Intervention | Experimental | Not described Receives: Dietary Supplement: 5-methyltetrahydrofolate (5-MTHF) |
| Placebo | Placebo comparator | Not described Receives: Dietary Supplement: placebo group |
What is being measured
The primary outcome is the main question the study is designed to answer.
Serum folate level
Changes in serum folate level pre and post the 3-month intervention period.
Measured: 3 months
Serum homocysteine level
Changes in serum homocysteine level pre and post the 3-month intervention period.
Measured: 3 months
Expression of PPARα and TNFα genes
Changes in expression of PPARα and TNFα genes pre and post the 3-month intervention period.
Measured: 3 months
Secondary outcomes (14)
Liver biochemical parameters (ALT (alanine aminotransferase), AST (aspartate aminotransferase), and GGT (gamma-glutamyl transferase)
Changes in ALT, AST, and GGT pre and post the 3-month intervention period.
Measured: 3 months
The fibrosis-4 (FIB-4) index
Changes in FIB-4 index pre and post the 3-month intervention period. The Fibrosis-4 (FIB-4) index will be calculated using the following formula: FIB-4 = (Age [years] × AST [U/L]) / (Platelet count [10⁹/L] × √ALT [U/L]). FIB-4 values >1.3, indicate a greater likelihood of liver fibrosis.
Measured: 3 months
Quality of life using SF-36 (36-Item Short Form Health Survey) questionnaires
Changes in quality-of-life pre and post the 3-month intervention period. Health-related quality of life will be assessed using the validated 36-Item Short Form Health Survey (SF-36). The questionnaire evaluates eight health domains: physical functioning, role limitations due to physical health, bodily pain, general health, vitality, social functioning, role limitations due to emotional problems, and mental health. Scores for each domain will be transformed to a 0-100 scale according to the standard scoring algorithm, with higher scores indicating better health-related quality of life.
Measured: 3 months
Lipid profile (triglycerides, total cholesterol, LDL-C (low-density lipoprotein cholesterol), HDL-C (high-density lipoprotein cholesterol))
Changes in lipid profile (triglycerides, total cholesterol, LDL-C, HDL-C) pre and post the 3-month intervention period.
Measured: 3 monhs
Fasting blood glucose
Changes in fasting blood glucose pre and post the 3-month intervention period.
Measured: 3 months
Fasting serum insulin
Changes in fasting serum insulin pre and post the 3-month intervention period.
Measured: 3 months
QUICKI (quantitative insulin sensitivity check index)
Changes in QUICKI pre and post the 3-month intervention period. The quantitative insulin sensitivity check index (QUICKI) will be calculated as 1/[log(fasting insulin [µU/mL]) + log(fasting glucose [mg/dL])], higher values indicating greater insulin sensitivity.
Measured: 3 months
HOMA-IR (homeostatic model assessment of insulin resistance
Changes in HOMA-IR pre and post the 3-month intervention period. Insulin resistance will be assessed using the homeostatic model assessment of insulin resistance (HOMA-IR), calculated as fasting insulin (µU/mL) × fasting glucose (mg/dL) / 405, higher values indicating greater insulin resistance.
Measured: 3 months
Weight
Changes in weight pre and post the 3-month intervention period.
Measured: 3 months
Body Mass Index (BMI)
Changes in BMI pre and post the 3-month intervention period. Body mass index (BMI) will be calculated as weight (kg) divided by the square of height (m²) and expressed as kg/m².
Measured: 3 months
Waist circumference
Changes in waist circumference pre and post the 3-month intervention period.
Measured: 3 months
Waist-to-hip ratio (WHR)
Changes in WHR pre and post the 3-month intervention period. Waist-to-hip ratio (WHR) will be calculated by dividing waist circumference by hip circumference.
Measured: 3 months
Body composition (fat-free mass)
Changes in fat-free mass (%) pre and post the 3-month intervention period. Body composition will be determined using a bioelectrical impedance analyzer.
Measured: 3 months
Body composition (fat mass)
Changes in fat mass (%) pre and post the 3-month intervention period. Body composition will be determined using a bioelectrical impedance analyzer.
Measured: 3 months
Timeline
From the start of the study to its completion; the mark is today. Estimated dates are the sponsor’s plan.
- First posted31 Jul 2026
- Study start30 Sep 2026 (estimated)
- Primary completion30 Aug 2027 (estimated)
- Study completion30 Aug 2027 (estimated)
Resources
- Official record on ClinicalTrials.govThe registry entry this page is built from, with its full change history.
- Other trials for Nonalcoholic Fatty Liver DiseaseStudies of the same condition, open and under way.
- What the trial phases meanHow a treatment moves from first tests in people to approval.
More details
For doctors and researchers: how the study is designed, who runs it, and its registry IDs.
Study design
- Study type
- Interventional
- Phase
- Phase N/A
- Primary purpose
- Treatment
- Allocation
- Randomised
- Intervention model
- Parallel groups
- Masking
- Quadruple blind
- Who is masked
- Participant, Care provider, Investigator, Outcomes assessor
- Enrolment
- 44 participants (estimated)
Sponsor and investigators
- Lead sponsor
- Tabriz University of Medical Sciences
- Sponsor type
- University, hospital or other
- Responsible party
- Principal investigator, Fatemeh Tamjid, Tabriz University of Medical Sciences
- Investigators
- Bahram Pourghassem Gargari, Principal investigator, Tabriz University of Medical Sciences
IDs, dates and publications
- ClinicalTrials.gov ID
- NCT07740109
- Sponsor's protocol ID
- IRCT20100123003140N26
- Acronym
- MASLD
- Status last verified
- Jul 2026
- First submitted
- 22 Jul 2026
- Last updated
- 31 Jul 2026
- Results posted
- No
- Data monitoring committee
- Yes
- FDA-regulated drug
- No
- FDA-regulated device
- No
- Individual data sharing
- Yes
Study at a glance
From the registry record, last updated by the sponsor on 31 Jul 2026. Recruitment status is as the sponsor reported it: confirm with the study team.
- Official title
- The Effect of 5-methyltetrahydrofolate Supplementation on Serum Folate and Homocysteine Level and PPARα and TNFα Gene Expression in Patients With Metabolic Dysfunction-Associated Steatotic Liver Disease: a Double-blind, Parallel Randomized Controlled Trial Study
- ClinicalTrials.gov ID
- NCT07740109
- Acronym / sponsor ID
- MASLD / IRCT20100123003140N26
- Conditions
- Nonalcoholic Fatty Liver Disease, Nonalcoholic Fatty Liver Disease (NAFLD), MASLD - Metabolic Dysfunction-Associated Steatotic Liver Disease, MASLD, NAFLD, Metabolic Dysfunction-Associated Steatotic Liver Disease
- Keywords
- 5-methyltetrahydrofolate, homocysteine, MASLD, NAFLD, Nonalcoholic Fatty Liver Disease, Metabolic Dysfunction-Associated Steatotic Liver Disease
- Study type
- Interventional
- Recruitment status
- Not yet recruiting
- Enrolment
- 44 participants (estimated)
- Lead sponsor
- Tabriz University of Medical SciencesUniversity, hospital or other
- Study start
- 30 Sep 2026 (estimated)
- Primary completion
- 30 Aug 2027 (estimated)
- Study completion
- 30 Aug 2027 (estimated)
- First posted
- 31 Jul 2026 (actual)
- Results
- Not posted in the source record
Clinical summary
The research question as the sponsor registered it. This describes what the study plans to test, not what it found.
To determine the effect of MTHF supplementation on serum folate and homocysteine level, metabolic, nutritional status, liver function, and PPARα and TNFα gene expression in patients with MASLD
Detailed description
Metabolic dysfunction-associated steatotic liver disease (MASLD), formerly termed non-alcoholic fatty liver disease (NAFLD), is diagnosed via liver biopsy or imaging when steatosis is present in the absence of alcohol intake or other hepatic disorders. As the liver manifestation of metabolic syndrome, it commonly coexists with obesity, diabetes, dyslipidemia, hypertension, and related conditions. Global prevalence of MASLD continues to rise.
Evidence from an earlier systematic review and meta-analysis indicated that MASLD patients had significantly lower serum folate and higher homocysteine concentrations. Folate is an essential water-soluble B vitamin that occurs in multiple chemically related forms. Food folates are mainly reduced and polyglutamated, with 5-MTHF predominating in both the diet and systemic circulation. 5-MTHF does not require reduction by DHFR and can enter the bloodstream directly for use. Reduced folates act as methyl donors in one carbon metabolism, supporting cellular proliferation, homocysteine re-methylation to methionine, nucleic acid synthesis and methylation of DNA, RNA, proteins and phospholipids.
Experimental studies have demonstrated that diet-induced hyperhomocysteinemia promotes hepatic steatosis and liver injury and folate as a key regulator of homocysteine concentration, may exert hepatoprotective effects. Evidence suggests that folate may improve hepatic lipid metabolism by activating peroxisome proliferator-activated receptor alpha (PPARα) signaling and modulate the immune response and reduce inflammatory mediators. Nevertheless, no evidence on the effects of folate on PPARα and TNFα gene expression in MASLD patients exist. Moreover, PPARα gene expression is dysregulated in MASLD and related metabolic conditions; PPARα is highly expressed in the liver, skeletal muscle and brown adipose tissue, stimulates β-oxidation and suppresses fatty-acid synthesis. Although the effect of 5-MTHF supplementation on gene expression of PPARα and TNFα in MASLD patients has not been examined, evidence showed that folate can modulate PPARα and TNFα. As folate has been shown to affect lipid metabolism and inflammation, we hypothesized that 5-MTHF supplementation might regulate PPARα and TNFα expression in MASLD patients. This randomized, double-blind, placebo-controlled clinical trial will therefore be undertaken to determine the effects of 5-MTHF supplementation on serum levels of folate and homocysteine, and gene expression of PPARα and TNFα in MASLD patients.
Study design
- Primary purpose
- Treatment
- Allocation
- Randomised
- Intervention model
- Parallel groups
- Masking
- Quadruple blind (participant, care provider, investigator, outcomes assessor)
- Number of arms
- 2
| Arm | Type | Receives |
|---|---|---|
| Intervention | Experimental | Dietary Supplement: 5-methyltetrahydrofolate (5-MTHF) |
| Placebo | Placebo comparator | Dietary Supplement: placebo group |
Who may qualify
- Ages
- 18 to 50 years
- Sex
- All
- Healthy volunteers
- Not accepted
- Age groups
- Adult
Key inclusion criteria (4 listed)
- Adult men or women (18-50 years)
- Diagnosis of MASLD (grade 1 or 2 of steatosis confirmed by ultrasound)
- Body mass index (BMI) = 25-34.9 kg/m²
- Providing written informed consent
Key exclusion criteria (6 listed)
- Pregnancy, lactation, or plans to get pregnant during the next three months.
- Liver disease (viral hepatitis, autoimmune liver disease, cirrhosis, drug-induced hepatotoxicity, or alcoholic fatty liver disease), heart or renal failure, kidney stones, any neoplasia, inflammatory disease, hypothyroidism, hypercortisolism, or hypertension
- Taking drugs affecting glucose or lipid metabolism, folate supplements, anti-obesity medications, weight-loss diets, or dietary supplements
- Lifestyle factors known to impact folate status (current smoking, alcohol intake, recreational drug use)
- Pre-existing conditions affecting folate status (malabsorptive or inflammatory bowel diseases, active celiac disease, gastric bypass surgery, atrophic gastritis, epilepsy, advanced liver disease, kidney dialysis, type 1 or 2 diabetes mellitus, or sickle cell trait/anemia)
Eligibility is decided by the study team after screening; this overview does not establish that a patient qualifies. Full inclusion and exclusion criteria.
Interventions
| Intervention | Type | Arms |
|---|---|---|
| 5-methyltetrahydrofolate (5-MTHF)Patients in this group will receive 5-methyltetrahydrofolate tablets (800 mcg) once a day for 90 days. Tablets will be manufactured by Ashbal Chemi pharmaceutical company (Qfol, Ashbal Chemi Co., Tehran, Iran). | Dietary supplement | Intervention |
| placebo groupPatients in this group will receive placebo for 90 days. The placebo is corn starch/ cellulose and will be consumed once a day. Placebo tablets will be manufactured by Ashbal Chemi Co. (Tehran, Iran). | Dietary supplement | Placebo |
Primary outcome
Serum folate level
Time frame: 3 months
Changes in serum folate level pre and post the 3-month intervention period.
Serum homocysteine level
Time frame: 3 months
Changes in serum homocysteine level pre and post the 3-month intervention period.
Expression of PPARα and TNFα genes
Time frame: 3 months
Changes in expression of PPARα and TNFα genes pre and post the 3-month intervention period.
14 secondary outcomes
Liver biochemical parameters (ALT (alanine aminotransferase), AST (aspartate aminotransferase), and GGT (gamma-glutamyl transferase)
Time frame: 3 months
The fibrosis-4 (FIB-4) index
Time frame: 3 months
Quality of life using SF-36 (36-Item Short Form Health Survey) questionnaires
Time frame: 3 months
Lipid profile (triglycerides, total cholesterol, LDL-C (low-density lipoprotein cholesterol), HDL-C (high-density lipoprotein cholesterol))
Time frame: 3 monhs
Fasting blood glucose
Time frame: 3 months
Fasting serum insulin
Time frame: 3 months
QUICKI (quantitative insulin sensitivity check index)
Time frame: 3 months
HOMA-IR (homeostatic model assessment of insulin resistance
Time frame: 3 months
Weight
Time frame: 3 months
Body Mass Index (BMI)
Time frame: 3 months
Waist circumference
Time frame: 3 months
Waist-to-hip ratio (WHR)
Time frame: 3 months
Body composition (fat-free mass)
Time frame: 3 months
Body composition (fat mass)
Time frame: 3 months
Results and safety
This study is still under way. Results are not available in the source record yet.
Research links
- Official record on ClinicalTrials.govLast updated by the sponsor 31 Jul 2026.
- Individual participant dataSharing planned: Yes
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Referral and contact
AzTrials does not enrol participants or take referrals. Contact the study team or a site listed below; they decide who can take part.
Bahram Pourghassem Gargari, PhD. Professor at TUMS contact
Bahram Pourghassem Gargari principal investigator
Tabriz University of Medical Sciences
1 site in 1 country: see every site and its status.
Questions about this trial
What is this clinical trial studying?
A study testing 5-methyltetrahydrofolate (5-MTHF) in people with Nonalcoholic Fatty Liver Disease and Nonalcoholic Fatty Liver Disease (NAFLD), run by Tabriz University of Medical Sciences at 1 site in Iran.
The full registered description is in the About this study section and the official record.
Who may be eligible to take part?
The sponsor lists 4 inclusion and 6 exclusion criteria. Eligibility is decided by the study team after screening, so read the full criteria and contact a listed site.
Is this clinical trial currently recruiting?
The record says: opening soon, as of the sponsor's last update on 31 Jul 2026. Status can change between updates, so confirm with the study contact before making plans.
Where is the study taking place?
The record lists 1 site in Iran. Whether a site is taking part right now should be confirmed with that site. See every site.
What are the possible benefits and risks?
They depend on the study and the treatment, and no study can promise a benefit. Some participants in this study may receive a placebo. The study team explains the known risks during informed consent, before anyone joins; the record itself does not list them.
Are results available for this study?
This study is still under way. Results are not available in the source record yet.
How can a doctor refer a patient or contact the research team?
Contact Fatemeh Tamjid, MSc, PhD student (+989144755462, tamjid.f@tbzmed.ac.ir), or a site near the patient. AzTrials does not enrol participants or take referrals.
What does the primary outcome measure?
Serum folate level (time frame: 3 months). The study lists 3 primary outcomes in all. It is what the study was designed to measure, not a finding.
Where can I read the original study information?
Does a listing mean the treatment is approved or proven to work?
No. A listing means the study is registered. It does not show that the treatment is safe, works, or is approved by a regulator.
How often is this information updated?
AzTrials copies changed records from ClinicalTrials.gov every night and reloads the whole registry every week. The sponsor last updated this record on 31 Jul 2026; AzTrials last refreshed its copy on 26 Oct 2026.