PsP Ganoderma Lucidum Supplementation and Biomarker Changes in Smokers (PSPGL)

Protocol for a Randomized, Single-Blind, Controlled Clinical Trial Evaluating the Effect of Peptide Polysaccharide Supplementation on Serum Malondialdehyde, Interleukin-6, and Cotinine Levels

The goal of this randomized controlled clinical trial is to evaluate whether polysaccharide peptide (PsP) supplementation from Ganoderma lucidum can modulate biomarkers of oxidative stress, inflammation, nicotine exposure, and stress response in adults exposed to cigarette smoke. The study is conducted in adults aged 20-50 years with active or passive exposure to cigarette smoke.

The main questions it aims to answer are:

• Does PsP supplementation change serum malondialdehyde (MDA) and superoxide dismutase (SOD) levels over 8 weeks?
• Does PsP supplementation affect serum interleukin-6 (IL-6), nitric oxide (NO), cotinine, nicotinic acetylcholine receptor (nAChR), and cortisol levels?

Researchers will compare participants receiving PsP supplementation with those receiving a placebo to evaluate differences in changes in the specified biomarkers.

Participants will:

• Be randomly assigned to receive PsP capsules (500 mg/day) or placebo for 8 weeks
• Provide fasting blood samples at baseline and at the end of the intervention
• Maintain usual lifestyle habits while avoiding antioxidant supplements during the study period

Study Overview

• Status: Completed
• Conditions: Inflammation; Oxidative Stress; Cigarette Smoking
• Intervention / Treatment: Dietary supplement: PsP

Detailed Description

Cigarette smoke exposure is associated with increased oxidative stress and inflammatory responses, largely mediated by excessive production of reactive oxygen species. These processes contribute to elevated levels of lipid peroxidation products and proinflammatory cytokines, as well as measurable biomarkers of nicotine exposure and stress-related physiological responses. Individuals with active or passive exposure to cigarette smoke may therefore experience sustained biochemical alterations linked to oxidative and inflammatory burden.

Polysaccharide peptide (PsP) derived from Ganoderma lucidum is composed primarily of β-(1,3)/(1,6)-D-glucans and peptide fractions that have demonstrated antioxidant and immunomodulatory activity in experimental and preclinical studies. Despite growing interest in PsP as a functional nutritional supplement, clinical evidence evaluating its effects on oxidative stress and inflammation in smoke-exposed human populations remains limited.

This randomized, single-blind, placebo-controlled clinical trial is designed to assess the effects of PsP supplementation on selected serum biomarkers related to oxidative stress, inflammation, nicotine exposure, and stress response. Eligible participants are adults aged 20-50 years with documented active or passive exposure to cigarette smoke. Participants are randomly assigned in a 1:1 ratio to receive either PsP supplementation or a visually identical placebo for a period of 8 weeks.

Participants assigned to the intervention group receive PsP capsules at a total daily dose of 500 mg, administered as one 250 mg capsule twice daily. Each capsule contains standardized amounts of total polysaccharides, including β-glucans, and peptide fractions. The control group receives placebo capsules containing inert excipients with no known biological activity. All study products meet applicable pharmacopeial standards for quality and safety. Adherence to the intervention is monitored through capsule counts and participant-maintained compliance logs.

Fasting blood samples are collected at baseline and at the end of the intervention period. Serum is separated and stored under controlled conditions until analysis. Laboratory assessments are planned using validated analytical methods to quantify biomarkers of oxidative stress, antioxidant activity, inflammation, nicotine exposure, receptor-related parameters, and stress-related hormones.

The primary outcome measures include changes in serum malondialdehyde (MDA) and superoxide dismutase (SOD) concentrations from baseline to week 8. Secondary outcome measures include changes in serum interleukin-6 (IL-6), nitric oxide (NO), cotinine, nicotinic acetylcholine receptor (nAChR) levels, and cortisol concentrations over the same period.

The planned statistical analysis includes descriptive summaries of baseline characteristics and inferential analyses to compare changes in outcome measures between study groups and within groups over time. Appropriate statistical methods are prespecified to account for baseline variability. All analyses are conducted using two-tailed tests with a prespecified level of statistical significance.

• Study Type: Interventional
• Enrollment (Actual): 200
• Phase: Not Applicable

Contacts and Locations

Study Locations

• Indonesia
  • East Java
    • Malang, East Java, Indonesia, 65145
      • University of Brawijaya

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: Adult
• Accepts Healthy Volunteers: Yes

Description

Inclusion Criteria:

1. Male or female adults aged 18 to 60 years.
2. Has smoked regularly for at least the past 12 months.
3. Able and willing to provide written informed consent.
4. Agrees to maintain usual smoking habits during the study period.
5. Able to comply with study procedures and visits.

Exclusion Criteria:

1. Current use of antioxidant supplements, vitamins, herbal supplements, or mushroom- derived products within the last 4 weeks.
2. Diagnosis of chronic inflammatory disease, autoimmune disease, diabetes mellitus, cardiovascular disease, chronic kidney disease, chronic liver disease, or cancer.
3. Acute illness, infection, or fever within the last 14 days.
4. Regular use of anti-inflammatory drugs (NSAIDs, corticosteroids) within the last 4 weeks.
5. Participation in another clinical study within the past 30 days.
6. Pregnant or breastfeeding.
7. Known allergy to Ganoderma lucidum or mushroom-derived compounds.
8. Alcohol dependence or substance abuse other than tobacco.

Study Plan

How is the study designed?

Design Details

• Primary Purpose: Treatment
• Allocation: Randomized
• Interventional Model: Parallel Assignment
• Masking: Single

Number of Arms

2

Arms and Interventions

Participant Group / Arm	Intervention / Treatment
No Intervention: Smoker Control Group (No Supplementation); Participants in this arm are smokers who do not receive any investigational product. They continue with their usual daily habits without additional supplements or treatments. This group serves as the control for evaluating the effects of PsP Ganoderma lucidum.
Experimental: Smokers Receiving PsP Ganoderma lucidum Supplementation; Participants in this arm are smokers who receive PsP Ganoderma lucidum as the investigational intervention. They follow the assigned PsP supplementation regimen for the full study duration to assess its effects compared with the control group.	Dietary supplement: PsP; PsP is a freeze dried Ganoderma lucidum Polysaccharide Peptide extract from mycelium cell wall. Each capsule contains 250 mg of Polysaccharide Peptide, which it equal to 180 mg β-1,3/1,6-D-Glucan.

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Change in Serum Malondialdehyde (MDA) Levels	Serum MDA concentration (nmol/mL) will be measured to assess oxidative stress status in participants receiving PsP compared with the control group	Baseline to 8 weeks
Change in Serum Superoxide Dismutase (SOD) Levels	Serum SOD concentration (U/mL) will be measured to assess oxidative stress status in participants receiving PsP compared with the control group	Baseline to 8 weeks

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Change in Serum Interleukin-6 (IL-6) Levels	Serum IL-6 concentration (pg/mL) will be measured to evaluate the inflammatory response following PsP supplementation compared with control	Baseline to 8 weeks
Change in Serum Nitric Oxide (NO) Levels	Serum NO concentration (µmol/L) will be measured to evaluate inflammatory response following PsP supplementation compared with control	Baseline to 8 weeks
Change in Serum Cotinine Levels	Serum cotinine concentration (ng/mL) will be measured as an objective biomarker of tobacco exposure to assess changes during the study period	Baseline to 8 weeks
Change in Serum Nicotinic acetylcholine receptors (nAChRs) Levels	Serum nAChRs concentration (ng/mL) will be measured as an objective biomarker of tobacco exposure to assess changes during the study period	Baseline to 8 weeks
Change in Serum Cortisol Levels	Serum Cortisol concentration will be measured as an objective biomarker of tobacco exposure to assess changes during the study period	Baseline to 8 weeks

Collaborators and Investigators

• Sponsor: University of Brawijaya
• Investigators: Principal Investigator: Titin A Wihastuti, Professor, Brawijaya University

Publications and helpful links

General Publications

• Kumboyono K, Chomsy IN, Hakim AK, Sujuti H, Hariyanti T, Srihardyastutie A, Wihastuti TA. Detection of Vascular Inflammation and Oxidative Stress by Cotinine in Smokers: Measured Through Interleukin-6 and Superoxide Dismutase. Int J Gen Med. 2022 Sep 16;15:7319-7328. doi: 10.2147/IJGM.S367125. eCollection 2022.
• Kumboyono K, Nurwidyaningtyas W, Chomsy IN, Wihastuti TA. Early Detection of Negative Smoking Impacts: Vascular Adaptation Deviation Based on Quantification of Circulated Endothelial Activation Markers. Vasc Health Risk Manag. 2021 Mar 23;17:103-109. doi: 10.2147/VHRM.S296293. eCollection 2021.
• Wihastuti TA, Heriansyah T. The inhibitory effects of polysaccharide peptides (PsP) of Ganoderma lucidum against atherosclerosis in rats with dyslipidemia. Heart Int. 2017 Apr 12;12(1):e1-e7. doi: 10.5301/heartint.5000234. eCollection 2017 Jan-Dec.

Helpful Links

• Product information for PsP (Peptide Polysaccharide) Ganoderma lucidum supplement, including composition, manufacturing details, and general product background provided by the manufacturer.

Study record dates

Study Major Dates

• Study Start (Actual): September 1, 2024
• Primary Completion (Actual): October 26, 2024
• Study Completion (Actual): November 30, 2024

Study Registration Dates

• First Submitted: December 3, 2025
• First Submitted That Met QC Criteria: December 18, 2025
• First Posted (Actual): January 5, 2026

Study Record Updates

• Last Update Posted (Actual): January 5, 2026
• Last Update Submitted That Met QC Criteria: December 18, 2025
• Last Verified: December 1, 2025

More Information

Terms related to this study

• Keywords: Inflammation; Oxidative Stress; Smokers; Ganoderma lucidum; β-Glucan; Peptide Polysaccharide (PsP)
• Additional Relevant MeSH Terms: Pathologic Processes; Pathological Conditions, Signs and Symptoms; Behavior; Smoking; Tobacco Smoking; Tobacco Use; Inflammation; Cigarette Smoking
• Other Study ID Numbers: 12757/UN10.F17.10.4/TU/2024; 01884.4/UN10.A0501/B/KS/2025 (Other Grant/Funding Number: University of Brawijaya)

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: NO
• IPD Plan Description: Individual participant data (IPD) will not be shared because the study does not include a data-sharing plan in the ethics approval or informed consent. Data contain sensitive biomarker and smoking-related information that may pose a re-identification risk for participants. Therefore, IPD will not be made publicly available.

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No