The Effect of Fermented Grape Juice Consumption on Fibromyalgia Patients

Official title

Determination of the Effect of Fermented Grape Juice Consumption on Oxidative Stress and Inflammation Biomarkers and SIRT-1 Levels in Fibromyalgia Patients

Overview

The goal of this clinical trial is to learn if a traditional grape-based fermented drink called hardaliye helps treat fibromyalgia in adult women. We will also learn whether adding hardaliye to a personalized nutrition plan works better than using either one alone. The main questions are: Does diet + hardaliye improve antioxidant status and lower oxidative stress more than diet only or hardaliye only? Does diet + hardaliye lower inflammation (TNF-α, IL-6, hs-CRP) and raise SIRT1 levels? Do these changes relate to less pain and better symptoms (fibromyalgia impact, gut symptoms, sleep, mood)? Are there benefits for weight, waist size, blood pressure, and blood lipids? How the groups are compared: Researchers will compare three groups for 8 weeks: Diet + Hardaliye (personalized medical nutrition plan + one 200 mL bottle of hardaliye daily) Diet only (personalized medical nutrition plan) Hardaliye only (usual diet + one 200 mL bottle of hardaliye daily) Participants will: Drink one 200 mL bottle of hardaliye each day if assigned to Diet + Hardaliye or Hardaliye only Follow a personalized nutrition plan if assigned to Diet + Hardaliye or Diet only Visit the clinic at the start and end of the study, with brief check-ins around weeks 2, 4, and 6 Give blood samples before and after the 8-week period Complete short questionnaires on pain, fibromyalgia impact, gut symptoms, sleep, and mood Keep simple logs of daily drink intake and diet plan adherence, and report any side effects Who can join: Adult women (20-40 years) with doctor-diagnosed fibromyalgia and BMI 25.0-29.9 kg/m² who meet the study's health and medication criteria.

Detailed description

This is a single-center, parallel-group, randomized clinical trial with three intervention conditions implemented over an 8-week period. After screening and baseline procedures, participants are assigned in a 1:1:1 ratio using a computer-generated block randomization sequence prepared by an independent researcher not otherwise involved in study conduct. Allocation concealment is maintained using sequentially numbered, opaque, sealed envelopes opened only after confirming eligibility.

Due to the dietary nature of the interventions, participant-level blinding is not feasible. However, biospecimen handling and laboratory analyses are performed on coded (anonymized) samples; laboratory personnel remain blinded to group assignment and time point to minimize measurement bias.

Personalized medical nutrition therapy (MNT) is delivered using a standardized counseling framework with individualized targets. Resting metabolic rate is estimated using the Mifflin-St Jeor equation, and daily energy prescription is derived using physical activity level (PAL). Macro-distribution targets are standardized (10-20% protein, 45-60% carbohydrate, 25-30% fat), with saturated fat \<7% of energy and trans fat \<1% to align with guidance-based cardiometabolic risk control.

To reduce confounding from background polyphenol exposure, participants are instructed to avoid high-polyphenol red/purple fruits and vegetables according to protocol-defined lists; diet plans are structured as three main meals plus 2-3 snacks, and standardized exchange lists are provided to support adherence.

Hardaliye intervention (product handling and quality assurance): The study beverage is sourced from a traditional producer in Kırklareli and manufactured via controlled spontaneous fermentation without starter culture.

Hardaliye is dispensed in 200 mL bottles and distributed weekly under cold-chain conditions; participants are instructed to consume the beverage daily within a consistent intake window (16:00-16:30) to standardize exposure timing.

To ensure batch-to-batch consistency, each production batch undergoes predefined acceptance checks (°Brix, pH, and titratable acidity), and key compositional markers are measured post-bottling (total phenolics via Folin-Ciocalteu, total monomeric anthocyanins via pH-differential method, and antioxidant capacity via ABTS-based TEAC). Analytical repeatability is monitored (replicates/parallel runs; CV target \<10% with repeat testing if unmet).

Batches outside acceptance ranges (e.g., °Brix 20.5-21.8; pH 3.50-4.00; acidity 0.68-0.75 g/100 mL; and protocol-defined total phenolic/anthocyanin targets) are excluded from participant distribution.

Where relevant, chromatographic confirmation (HPLC-DAD) is used to verify the phenolic profile of the study product and support linkage of total phenolic estimates to compound-level composition.

Participants complete an initial screening visit followed by a baseline (Day 0) visit and an end-of-intervention (Week 8) visit, with brief interim check-ins at Weeks 2, 4, and 6 for adherence review and systematic safety monitoring. Interim contacts include beverage count/log review, reinforcement of dietary counseling where applicable, and adverse event solicitation/documentation.

Adherence is operationalized as ≥80% compliance with the assigned beverage intake and/or nutrition therapy targets; intake and adherence are captured using simple participant logs reviewed at each contact.

Dietary intake is assessed using structured food records collected at multiple time points (7-day records at baseline, Week 4, and Week 8; and additional 3-day records during specified weeks) to quantify energy and nutrient intake and to support adherence monitoring. Nutrient analyses are performed using BeBIS software and interpreted against national reference intakes.

Dietary polyphenol exposure is estimated using Phenol-Explorer (v3.6) to derive total polyphenols and relevant subclasses; an antioxidant diet quality score (DAQS) is calculated based on intake of key antioxidant nutrients. Physical activity is monitored via 24-hour recall on diary days, and PAL is derived using activity-specific PAR values combined with Mifflin-St Jeor BMR estimates to support stability of lifestyle exposure during follow-up.

Fasting venous blood is collected at baseline and Week 8 and processed according to institutional standard operating procedures (timing, centrifugation, aliquoting, storage). Samples are labeled with coded identifiers to preserve analytical blinding, and assays are performed following validated methods and kit instructions with internal QC steps documented prospectively.

Data are captured in structured case report forms and reconciled against visit checklists, distribution records, and participant logs. Primary analyses follow an intention-to-treat approach. Group differences in change over time are evaluated using repeated-measures modeling (e.g., linear mixed-effects models including group, time, and group×time interaction), with prespecified sensitivity analyses based on adherence and protocol deviations; missing-data handling and multiplicity procedures are defined in the statistical analysis plan.

Interventions

• Behavioral Medical Nutrition Therapy (MNT)
  Individually tailored medical nutrition therapy delivered using a standardized counseling framework, energy prescription derived from predictive equations and physical activity level, with standardized macronutrient targets and follow-up reinforcement per protocol.
• Other Hardaliye (200 mL/day)
  Traditional grape-based fermented beverage dispensed in 200 mL bottles; participants consume 200 mL once daily for the intervention period with standardized intake instructions and product handling.

Primary outcome measures

• Fibromyalgia Impact Questionnaire Revised [Time frame: Baseline to Week 8]
• Pain Severity (Visual Analog Scale, VAS) [Time frame: Baseline to Week 8]
• Gastrointestinal symptoms (Gastrointestinal Symptom Rating Scale, GSRS) [Time frame: Baseline to Week 8]
• Sleep Quality (Pittsburgh Sleep Quality Index, PSQI) [Time frame: Baseline to Week 8]
• Depressive Symptoms (Beck Depression Inventory, BDI) [Time frame: Baseline to Week 8]

Secondary outcome measures (7)

• Total Antioxidant Status (TAS; μmol Trolox eq/L) [Time frame: Baseline to Week 8]
• Total Oxidant Status (TOS; μmol H₂O₂ eq/L) [Time frame: Baseline to Week 8]
• Oxidative Stress Index (OSI = TOS×100 / TAS) [Time frame: Baseline to Week 8]
• Plasma Malondialdehyde (MDA; TBARS method) [Time frame: Baseline to Week 8]
• Serum 8-Hydroxy-2'-deoxyguanosine (8-OHdG) [Time frame: Baseline to Week 8]
• Inflammatory biomarkers (TNF-α, IL-6, hs-CRP) [Time frame: Baseline to Week 8]
• Serum Sirtuin-1(SIRT1) [Time frame: Baseline to Week 8]

Eligibility criteria

Inclusion criteria

• Female, 20-40 years
• BMI 25.0-29.9 kg/m²
• FM per 2016 ACR criteria; abdominal pain/cramp per SSS sub-item
• MEDAS ≤5 at screening.
• Willing to maintain habitual activity and comply with assigned intervention; able to provide informed consent.

Exclusion criteria

• Age <20 or >40
• BMI ≤24.99 or ≥30.0 kg/m²
• Professional athletes; night-shift workers
• Food allergies; acute infection; antibiotic use within past month or during intervention.
• Use (past 3 months) of dietary supplements; recent weight-loss or special diets (e.g., ketogenic).
• Pregnancy/lactation; menopause.
• Major hepatic/renal/cardiac/immune disease; chronic GI diseases; diabetes; CNS disorders; cancer; thyroid disease; severe pulmonary disease. Inflammatory arthritis/autoimmune disease impacting weight; medications including corticosteroids, estrogens, analgesics/anti-inflammatories, anti-diabetic or lipid-lowering drugs.
• Severe psychiatric illness/substance use; heavy alcohol/smoking.
• High recent intake of grape juice/molasses; excess tea/coffee (≥7 cups/day).
• Adherence <80% to beverage or MNT during run-in/screening.

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

Healthy volunteers: No

Study design

Allocation

Randomized

Model

Parallel assignment

Masking

Open label

Primary purpose

Treatment

Study locations

Turkey (Türkiye) · 1 center

• İstanbul University-Cerrahpaşa Hospital — Istanbul

Publications

• Erel O. A new automated colorimetric method for measuring total oxidant status. Clin Biochem. 2005 Dec;38(12):1103-11. doi: 10.1016/j.clinbiochem.2005.08.008. Epub 2005 Oct 7. PMID 16214125
• Erel O. A novel automated method to measure total antioxidant response against potent free radical reactions. Clin Biochem. 2004 Feb;37(2):112-9. doi: 10.1016/j.clinbiochem.2003.10.014. PMID 14725941
• Al-Aubaidy HA, Jelinek HF. Oxidative stress and triglycerides as predictors of subclinical atherosclerosis in prediabetes. Redox Rep. 2014 Mar;19(2):87-91. doi: 10.1179/1351000213Y.0000000080. Epub 2014 Jan 13. PMID 24520969
• Jentzsch AM, Bachmann H, Furst P, Biesalski HK. Improved analysis of malondialdehyde in human body fluids. Free Radic Biol Med. 1996;20(2):251-6. doi: 10.1016/0891-5849(95)02043-8. PMID 8746446
• Gerrior S, Juan W, Basiotis P. An easy approach to calculating estimated energy requirements. Prev Chronic Dis. 2006 Oct;3(4):A129. Epub 2006 Sep 15. PMID 16978504
• Bekar C, Goktas Z. Validation of the 14-item mediterranean diet adherence screener. Clin Nutr ESPEN. 2023 Feb;53:238-243. doi: 10.1016/j.clnesp.2022.12.026. Epub 2022 Dec 29. PMID 36657918
• Martinez-Gonzalez MA, Garcia-Arellano A, Toledo E, Salas-Salvado J, Buil-Cosiales P, Corella D, Covas MI, Schroder H, Aros F, Gomez-Gracia E, Fiol M, Ruiz-Gutierrez V, Lapetra J, Lamuela-Raventos RM, Serra-Majem L, Pinto X, Munoz MA, Warnberg J, Ros E, Estruch R; PREDIMED Study Investigators. A 14-item Mediterranean diet assessment tool and obesity indexes among high-risk subjects: the PREDIMED tr PMID 22905215
• Rivas A, Romero A, Mariscal-Arcas M, Monteagudo C, Lopez G, Lorenzo ML, Ocana-Peinado FM, Olea-Serrano F. Association between dietary antioxidant quality score (DAQs) and bone mineral density in Spanish women. Nutr Hosp. 2012 Nov-Dec;27(6):1886-93. doi: 10.3305/nh.2012.27.6.6039. PMID 23588435

Identifiers

NCT: NCT07403929 · 1422809

Primary sources (government registries)

• ClinicalTrials.gov (US NIH)

View this study on ClinicalTrials.gov ↗