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INDONESIA
The Indonesian Biomedical Journal
Published by The Prodia Education and Research Institute
ISSN : -     EISSN : -     DOI : -
Core Subject : Health, Science,
Biochemistry, Genetics & Molecular Biology Public Health
Arjuna Subject : -
Articles 10 Documents
 1 
Search results for , issue "Vol 17, No 4 (2025)" : 10 Documents clear
High-yield and Functional IgY Anti-S1 SARS-CoV-2 Delta Variant from Chicken Immunized with In-house Recombinant Protein Zarkasie, Kamaluddin; Ginting, Teridah Ernala; Poeloengan, Andrea Hynan; Andriani, Febi; Lages, Aksar Chair; Cornelia, Vina; Nugraha, Anggah; Rosidi, Bustanur; Indrasari, Setyarina; Nidom, Chairul Anwar; Yusuf, Irawan
The Indonesian Biomedical Journal Vol 17, No 4 (2025)
Publisher : The Prodia Education and Research Institute (PERI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18585/inabj.v17i4.3693

Abstract

BACKGROUND: During COVID-19 pandemic, effective preventive and therapeutic strategies was urgently necessary, especially with the emerging Delta variants. Previous existing interventions have some limitations, and passive immunization using immunoglobulin Y (IgY) is considered viable. However, the potential of IgY antibodies remains underexplored locally. Therefore, a feasible study was conducted to produce IgY against the SARS-CoV-2 Delta variant from chickens raised on a farm, instead of a laboratory environment, by using in-house recombinant protein, to offer a promising low-cost alternative for passive immunization.METHODS: In-house HEK293 cell line was produced by secreting recombinant S1 subunit protein of SARS-CoV-2 Delta variant. The protein was confirmed by western blot and used as an immunogen in two chickens. IgY was extracted from egg yolks. Titers and neutralization activity of IgY against SARS-CoV-2 Delta variant were confirmed by enzyme-linked immunosorbent assay (ELISA) and surrogate virus neutralization test (sVNT). The functionality of IgY was then tested on lysates from COVID-19 patients' nasopharyngeal swabs with dot blot method.RESULTS: The high anti-S1 IgY titers and neutralization activity was confirmed following immunization with 1 mg immunogen at 2–3-week intervals. IgY titers varied between chickens and time points. The mean titers showed significant increase after the fifth immunization (1.21, 1.72 and 1.48; with p<0.05 for all). Neutralization activity appeared after the second immunization and was significant after the third immunization (31.9%, 34.95%, 26.9%, 47.6%, 54.95% and 57.3%; with p<0.05 for all). The results of this study showed that the extracted IgY reacted to COVID-19 patients' nasopharyngeal swabs lysates.CONCLUSION: A high-yield and functional IgY anti-S1 SARS-CoV-2 Delta variant can be produced from chicken immunized with an in-house recombinant protein. This can be an alternative for affordable and effective IgY production during public health emergencies.KEYWORDS: IgY, SARS-CoV-2, S1 subunit protein, egg yolk, chicken
Non-Invasive Prenatal Testing with Next Generation Sequencing Methods in Birth Defect Pregnancy: A Pilot Study Suardika, Anom; Kusuma, Anak Agung Ngurah Jaya; Ermayanti, Ni Gusti Ayu Manik; Widiyanti, Endang Sri; Wibawa, I Gusti Ngurah Agung Satria; Silvana, Divika; Budayasa, Anak Agung Gede Raka; Dewi, Ni Nyoman Ayu; Jawi, I Made; Sun, H. Sunny; Tang, Yen-An
The Indonesian Biomedical Journal Vol 17, No 4 (2025)
Publisher : The Prodia Education and Research Institute (PERI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18585/inabj.v17i4.3753

Abstract

BACKGROUND: Identification of cell-free foetal DNA (cffDNA) in maternal blood, combined with next-generation sequencing (NGS) advancement, has paved the way for non-invasive prenatal screening to detect foetal aneuploidies. However, there is limited evidence on its diagnostic accuracy when compared with gold-standard invasive tests specifically in pregnancies complicated by birth defects in Indonesia. This study was conducted to evaluate the precision of non-invasive prenatal testing (NIPT) using NGS and ultrasound findings compared with the established benchmarks of amniocentesis and neonatal karyotyping through G-banding analysis, which is an invasive procedures, in a private laboratory setting for pregnancies with birth defect.METHODS: An observational cohort study involving pregnant women with foetal birth defects in central nervous system, facial, heart, gastrointestinal tract, urinary tract abnormalities and suspected Down Syndrome was conducted. The foetal birth defects were identified in the first trimester with ultrasound screening. Venous blood was drawn from the mother for NGS-based NIPT examination. As a gold standard, amniocentesis or neonatal G-banding karyotyping was conducted.RESULTS: Using G-banding karyotyping as gold standard, the results indicated that NIPT using the NGS method and ultrasound findings achieved 100% sensitivity, 100% specificity, and 100% accuracy in detecting trisomy 13, 18, and 21, as well as foetal sex chromosome abnormalities. Additionally, a case of tetrasomy 9p was identified through G-banding karyotyping, which was associated with multiple clinical abnormalities.CONCLUSION: NIPT with NGS methods and ultrasound findings demonstrated 100% accuracy for the screening of trisomy 13, 18, and 21 in birth defect pregnancy, which is comparable with G-banding analysis as a gold standard. Therefore, this suggest that these approaches offer a safe early detection, highly accurate alternative in high risk setting, compared to invasive procedure in Indonesia where access to such testing may be limited. KEYWORDS: G-banding karyotyping, next generation sequencing, non-invasive prenatal testing
Therapeutic Potential of Gut Microbiota in Hypertension: Mechanisms of Immune Modulation and Inflammation Meiliana, Anna; Dewi, Nurrani Mustika; Wijaya, Andi
The Indonesian Biomedical Journal Vol 17, No 4 (2025)
Publisher : The Prodia Education and Research Institute (PERI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18585/inabj.v17i4.3565

Abstract

Emerging evidence links gut dysbiosis to numerous ailments, including hypertension and metabolic diseases. Multi-omics techniques have revealed that hypertensive individuals exhibit distinct alterations in their gut bacterial composition and metabolite profiles. The gut microbiome influences blood pressure through several mechanisms. For instance, microbiota-derived metabolites can have beneficial effects, such as those from short-chain fatty acids (SCFAs), or detrimental ones, like trimethylamine N-oxide (TMAO). These molecules modulate downstream signaling pathways via G protein-coupled receptors or direct immune cell activation. Furthermore, dysbiosis can compromise the gut epithelial barrier, leading to systemic inflammation that activates key regulatory pathways like the renin-angiotensin-aldosterone system (RAAS), the autonomic nervous system, and the immune system. Given these connections, the gut microbiome is a promising therapeutic target for hypertension. This review explores the potential of modulating the gut microbiota to manage blood pressure, focusing on the underlying mechanisms of immune modulation, inflammation, and microbial metabolites. By focusing on the 'how' rather than the 'what' of hypertension, it is identified that immune-mediated inflammation is orchestrated by the gut microbiota, as the core mechanism driving the disease. Gut dysbiosis is triggered by environmental factors like high-salt diets, perpetuates a pro-inflammatory state that undermines the efficacy of conventional antihypertensive drugs and contributes to treatment-resistant hypertension. Consequently, modulating the gut microbiota through targeted interventions, including dietary fiber, probiotics, and fecal transplantation, might represents a critical evolution in treatment. This approach moves beyond managing symptoms to directly correcting the inflammatory dysfunction at the heart of the disease, offering a powerful strategy to complement existing therapies.KEYWORDS: hypertension, inflammation, gut microbiota, metabolite
FOXO1 and FYN Expression Trends in Breast Cancer Stem Cells: An Integrative Study of Single Nucleotide Polymorphism (SNP) Array and Quantitative PCR (qPCR) Analysis Margaret, Ay Ly; Wanandi, Septelia Inawati; Fadilah, Fadilah; Paramita, Rafika Indah
The Indonesian Biomedical Journal Vol 17, No 4 (2025)
Publisher : The Prodia Education and Research Institute (PERI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18585/inabj.v17i4.3656

Abstract

BACKGROUND: Currently, identification of breast cancer stem cells (BCSCs) commonly relies on CD24-/CD44+ expression profiles. However, few studies have integrated genomic mutation data with experimental gene expression validation in CSC and non-CSC populations. Genotyping results of CD24-/CD44+ MDAMB-231 cells revealed 36 mutations in BCSCs compared to non-BCSCs, with upregulated FOXO1 and FYN that might represent promising candidate biomarkers for this subpopulation. Therefore, in this study, single nucleotide polymorphism (SNP) and quantitative polymerase chain reaction (qPCR) analysis were performed to assess the association between mutations and expression trends of FOXO1 and FYN in MDAMB-231 cell, as breast cancer cell model with stem-like traits and well-characterized profile.METHODS: Genomic DNA was isolated from BCSC and non-BCSC DNA from the MDAMB-231 cell line. Mutation analysis was conducted using PLINK, while gene expressions of FOXO1 and FYN were quantified by one-step SYBR Green-based qPCR, using 18s rRNA as a reference. Data was then analyzed with the Livak (2−∆∆Ct) method.RESULTS: Among 36 mutations found in BCSCs of the MDAMB-231 cell line, PTEN (rs786204914) and CHEK2 (rs587782401) were identified as pathogenic. While FOXO1 (2.989±2.817 vs. 1.072±0.388) and FYN (1.405±0.072 vs. 0.855±0.140) mRNA levels were found to be higher in CSCs compared to non-CSCs, though these differences was not statistically significant.CONCLUSION: Pathogenic mutations in CHEK2 and PTEN were detected within BCSC population, implying a potential influence on the expression of FOXO1 and FYN, though not statistically significant. These findings suggest a possible, but as yet unverified, association between gene mutations and expression patterns, emphasizing the importance of further functional studies to validate FOXO1 and FYN as biomarkers for BCSCs.KEYWORDS: breast cancer stem cells, FOXO1, FYN, PTEN, CHEK2, mutation, biomarker
Early Active Exercise Improves MDA, SOD, and GSH Levels without Memory or NO Changes in Wistar Rats Fairof, Muhammad Hafiz Zuhdi; Mat Ludin, Arimi Fitri; Anas, Nur Diyana; Husna, Ainaa; Che Hasan, Ahmad Muhaimin; Ahmad Abdullah, Amirul Hafiz; Ibrahim, Farah Wahida; Pramono, Adriyan; Rajab, Nor Fadilah
The Indonesian Biomedical Journal Vol 17, No 4 (2025)
Publisher : The Prodia Education and Research Institute (PERI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18585/inabj.v17i4.3663

Abstract

BACKGROUND: Circadian rhythms regulate various physiological processes, including responses to exercise. However, the effects of exercise timing on cognitive function and oxidative stress remain unclear. One key factor in oxidative stress is nitric oxide (NO), an enzyme complex that produces reactive oxygen species (ROS) as part of normal cellular signaling. Excessive NO activity can disrupt redox balance and contribute to neuronal damage. An imbalance favoring oxidative stress can impair memory and learning, while a higher antioxidant capacity supports brain health and cognitive performance. This study was performed to investigate whether early active and late active aerobic exercise differentially impact cognitive function and oxidative stress biomarkers in Wistar rats.METHODS: Sixteen male Wistar rats were randomly assigned to four groups: early active control, late active control, early active exercise, and late active exercise. The exercise groups underwent treadmill running for seven weeks, five days per week. Cognitive performance was assessed using the novel object recognition (NOR) test, while oxidative stress biomarkers, including malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione (GSH) were analyzed from brain tissue samples (hippocampus) following already established methods. Meanwhile the NO were assessed using Enzyme-linked Immunosorbent Assay (ELISA).RESULTS: This study showed that exercise timing did not significantly affect non-spatial memory performance. However, early active exercise led to a significant increase in SOD and GSH levels compared to the control and late active exercise groups, suggesting enhanced antioxidant activity. Conversely, late active exercise did not significantly impact oxidative stress markers. No changes was found in the NO concentration in both exercise timing.CONCLUSION: These findings suggest that exercise performed during the early active phase may be more beneficial for oxidative stress regulation, potentially contributing to long-term cognitive resilience.KEYWORDS: circadian rhythm, exercise timing, cognitive function, oxidative stress
Insulin-like Growth Factor-1 and Calcium Ion Levels are Negatively Associated with Serum β-Cross Laps in Multi-transfused β-thalassemia Major Patients Widodo, Sabar; Widyastiti, Nyoman Suci; Limijadi, Edward Kurnia Setiawan; Hendrianingtyas, Meita; Retnoningrum, Dwi; Ariosta, Ariosta; Nency, Yetty Movieta
The Indonesian Biomedical Journal Vol 17, No 4 (2025)
Publisher : The Prodia Education and Research Institute (PERI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18585/inabj.v17i4.3706

Abstract

BACKGROUND: Thalassemia patients with repeated transfusions (multi-transfusion) are often at risk of experiencing early osteoporosis. Several studies have demonstrated that osteoporosis in these individuals was associated with altered bone remodeling, characterized by decreased insulin-like growth factor-1 (IGF-1) and serum calcium ions, as well as increased serum β-Cross Laps (β-CTx) levels. Despite the prevalence of this condition, there is limited literature on the relationship between IGF-1 levels and calcium ions with β-CTx. Therefore, this study was conducted to examine the relationship between IGF-1 and calcium ions levels with serum β-CTx in multi-transfused β-thalassemia major patients.METHODS: A cross sectional study involved 29 thalassemia patients with multiple transfusions, aged 2-18 years, that were selected from the electronic medical records. Calcium ion levels were examined using ion selective electrode method. Subsequently, IGF-1 and serum β-CTx levels were examined by enzyme-linked immunosorbent assay (ELISA), and the data were analyzed using the Pearson correlation test.RESULTS: The results showed that the mean serum IGF-1 levels, calcium ion, and β-CTx were 20.11±20.763 ng/mL, 1.26±0.07 mmol/L, and 9330.40±1696.76 ng/mL, respectively. Statistical analysis showed a significant relationship between the levels of IGF-1 (r=-0.573; p=0.001) and calcium ion (r=-0.373; p=0.046) with serum β-CTx. A moderate negative relationship was found between IGF-1 levels and β-CTx, while calcium ion levels and β-CTx showed a weak negative relationship.CONCLUSION: A moderate negative correlation between IGF-1 and serum β-CTx, and a weak negative correlation between calcium ion and serum β-CTx suggest that IGF-1 and calcium ions may serve as potential indicators of bone turnover and osteoporosis risk in multi-transfused β-thalassemia major patients, underscoring their potential role in routine clinical evaluations.KEYWORDS: miR-200a expression, NO level, early-onset preeclampsia, late-onset preeclampsia
CYP2C9 rs1057910 Genotype and Its Association with Paraclinical Characteristics in Gout Patients in the Northeast Region of Vietnam Vu, Lan Thi; Hoang, Yen Thi Thu; Nguyen, Yen Thi; Nguyen, Vien Van; Nguyen, Hien Thu; Do, Thanh Ha; Hoang, Tuan Phan
The Indonesian Biomedical Journal Vol 17, No 4 (2025)
Publisher : The Prodia Education and Research Institute (PERI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18585/inabj.v17i4.3713

Abstract

BACKGROUND: Celecoxib and lesinurad are medications used in the management of gout, and their metabolism is significantly influenced by genetic variations in the CYP2C9 enzyme. CYP2C9*3 (rs1057910) is associated with reduced CYP2C9 activity. This study investigated the association between CYP2C9 rs1057910 genotype and paraclinical characteristics in gout patients from the Northeast region of Vietnam.METHODS: A total of 139 gout patients were recruited and their paraclinical characteristics including red blood cell, hemoglobin, hematocrit, white blood cell, neutrophil, lymphocyte, platelet, glucose, urea, creatinine, uric acid, triglyceride, total cholesterol, high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C) were collected. The CYP2C9 rs1057910 genotypes were identified by Sanger sequencing method of PCR products, analyzed with BioEdit software, and verified using the NCBI dbVar database. Statistical analyses were performed using SPSS.RESULTS: The cohort was predominantly male (93.5%), with female patients showing a significantly higher mean age (70.33±10.64 years) than males (51.81±14.93 years, p<0.001). This study showed significant positive correlations between uric acid concentration, creatinine (r=0.201, p=0.018) and platelet count (r=0.169, p=0.046). The wild-type homozygous CYP2C9*1/*1 genotype was found in 92.09% of patients; the CYP2C9*1/*3 and CYP2C9*3/*3 genotypes were identified in 7.19% and 0.72%, respectively. No significant differences in most paraclinical parameters were observed between genotype groups, except for HDL-C levels, which were significantly higher in CYP2C9*3 carriers (p=0.000).CONCLUSION: This study showed that the CYP2C9*3 carrier is significantly associated with higher HDL-C levels compared to the CYP2C9*1/*1 in gout patients. This finding suggests that the CYP2C9*3 variant may influence lipid metabolism in a way that promotes a more favorable lipid profile, which are considered protective against cardiovascular disease.KEYWORDS: CYP2C9 gene, CYP2C9*3, CYP2C9 rs1057910 genotype, gout patients, paraclinical characteristics
Hypomethylation of the Soluble Fms-like Tyrosine Kinase 1 (sFlt-1) Gene Promoter Region and Elevated sFlt-1 Placental Expression as Risk Factors for Preeclampsia Kusuma, Anak Agung Ngurah Jaya; Darmayasa, I Made; Putra, I Gede Mega; Suardika, Anom; Pangkahila, Evert Solomon; Duarsa, Vidya Saraswati Putri; William, William
The Indonesian Biomedical Journal Vol 17, No 4 (2025)
Publisher : The Prodia Education and Research Institute (PERI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18585/inabj.v17i4.3744

Abstract

BACKGROUND: Preeclampsia significantly contributes to maternal and fetal morbidity and mortality worldwide, marked by an imbalance of angiogenic factors, particularly increased soluble Fms-like tyrosine kinase-1 (sFlt-1), leading to endothelial dysfunction. Epigenetic regulation, including DNA methylation of the sFlt-1 promoter, has been suggested to influence sFlt-1 expression, but the data in Indonesian population are limited. This study was perfmed to determine whether hypomethylation of the sFlt-1 promoter and elevated placental sFlt-1 expression are associated with increased risk of preeclampsia.METHODS: A case-control study was conducted involving 30 women with preeclampsia and 30 normotensive pregnant women. Subjects were selected based on eligibility criteria that included singleton pregnancy and gestational age of ≥37 weeks. DNA methylation of the sFlt-1 promoter was assessed using methylation-specific polymerase chain reaction (PCR), and sFlt-1 expression was measured by enzyme-linked immunosorbent assay (ELISA). Statistical analyses, including Mann-Whitney U, Chi-square tests, Receiver-operating characteristic (ROC) curve analysis, and multivariate logistic regression, were performed to evaluate the relationship between methylation levels, gene expression, and preeclampsia risk.RESULTS: The preeclampsia group had significantly lower methylation levels of sFlt-1 promoter and higher placental sFlt-1 expression (both p<0.001). Hypomethylation of sFlt-1 promoter (adjusted odd ratio (AOR): 21.18; 95% CI: 2.49–179.72; p=0.005), high sFlt-1 expression (AOR: 12.55; 95% CI: 1.95–80.83; p=0.008), and obesity (AOR: 11.15; 95% CI: 2.01–61.78; p=0.006) were identified as independent risk factors for preeclampsia.CONCLUSION: Hypomethylation of sFlt-1 promoter and elevated placental sFlt-1 expression are significant independent risk factors for preeclampsia. These findings suggest that hypomethylation of sFlt-1 promoter and elevated placental sFlt-1 expression may serve as potential epigenetic biomarkers for early detection and targeted intervention in preeclampsia.KEYWORDS: preeclampsia, sFlt-1, gene expression, hypomethylation, placenta, risk factor
GSH-conjugation Reduces PGV-1 Cytotoxicity and Its Ability in Downregulating N-Myc, β-catenin, and p62 Protein in Huh-6 Cells Utomo, Rohmad Yudi; Meiyanto, Edy; Susidarti, Ratna Asmah
The Indonesian Biomedical Journal Vol 17, No 4 (2025)
Publisher : The Prodia Education and Research Institute (PERI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18585/inabj.v17i4.3634

Abstract

BACKGROUND: Pentagamuvone-1 (PGV-1), a synthetic curcumin analogue, exhibits potent anticancer activity against Hepatocellular Carcinoma (HCC) by disrupting cell cycle regulation and downregulating key oncogenes such as N-Myc. Numerous studies have examined the role of glutathione (GSH) conjugation in modulating the anticancer properties of curcumin and its analogues. In contrast, the impact of PGV-1 metabolism, particularly GSH conjugation, and its implications for anticancer efficacy have not yet been elucidated. This study was performed to prepare GSH-conjugated PGV-1 (PGV-1-(GSH)2) as the model of PGV-1 metabolite and evaluate its potential distinct cytotoxicity on Huh-6 cells.METHODS: PGV-1 was synthesized via an acid-catalyzed reaction between 4-hydroxy-3,5-dimethylbenzaldehyde and cyclopentanone while PGV-1-(GSH)2 was obtained through reflux at 70oC for 2 hours. The cytotoxic effects of PGV-1 and PGV-1-(GSH)2 on Huh-6 and JHH4, two HCC cells, were assessed using a cell counting kit-8 (CCK-8) assay, while immunoblotting was performed to evaluate their impact on N-Myc and its downstream protein such as β-catenin, and p62.RESULTS: PGV-1-(GSH)2 was prepared through GSH conjugation of PGV-1 in orange color solution, as confirmed by Electrospray Ionization Mass Spectrometry (ESI-MS), Fourier Transform Infrared Spectroscopy (FT-IR), and Nuclear Magnetic Resonance (NMR) analysis. Cytotoxicity assays revealed that PGV-1-(GSH)2 exhibited less potent anticancer activity against HCC cells than PGV-1. GSH conjugation also decreased the ability of PGV-1 in downregulating the N-Myc, β-catenin, and p62 protein level.CONCLUSION: The prepared PGV-1-(GSH)2 reduces the cytotoxicity of PGV-1 and its ability on downregulating N-Myc, β-catenin, and p62 in Huh-6 cells. These findings highlight the need for further exploration about the study of PGV-1 metabolism which could affect the anticancer efficacy against HCC.KEYWORDS: curcumin, PGV-1, GSH, HCC, N-Myc
Renoprotective Effects of Hydroxychloroquine and Folinic Acid via ET-1 and NLRP3 Modulation in Reducing Tubular Injury in A Rabbit Model of Methanol-induced Acute Kidney Injury Prayuda, Prayuda; Widiana, I Gde Raka; Suega, Ketut; Kandarini, Yenny; Winarti, Ni Wayan; Purwanto, Bambang
The Indonesian Biomedical Journal Vol 17, No 4 (2025)
Publisher : The Prodia Education and Research Institute (PERI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18585/inabj.v17i4.3747

Abstract

BACKGROUND: Methanol intoxication is associated with significant morbidity and mortality, particularly when acute kidney injury (AKI) developed. Emerging evidence implicates Endothelin-1 (ET-1) and Nucleotide-binding domain leucine-rich repeat-containing pyrin receptor 3 (NLRP3) inflammasome in renal injury, but their roles in methanol-induced AKI remain unclear. To date, no studies have examined whether hydroxychloroquine or folinic acid, which are known to modulate ET-1 and NLRP3 signaling, could mitigate renal injury in this setting. This study evaluated their therapeutic effects in a rabbit model of methanol-induced AKI.METHODS: The animals subjects were randomly assigned to four groups: control group receiving aquabidest, folinic acid group receiving 2 mg/kg body weight (BW) intraperitoneal folinic acid, hydroxychloroquine group receiving 30 mg/kg BW oral hydroxychloroquine phosphate, and combination group receiving both folinic acid and hydroxychloroquine at the same dosages. Histopathological evaluation of tubular injury scores and immunohistochemical analysis of ET-1 and NLRP3 expression were then conducted.RESULTS: Expressions of ET-1, NLRP3, and tubular injury scores were significantly lower in the hydroxychloroquine, folinic acid, and combination therapy groups compared to the control group (p<0.001). Expression of ET-1 was lowest in folinic acid group (59.38±0.71%), followed by combination group (62.23±1.98%) and hydroxychloroquine group (62.43±1.81%), compared to control group (72.14±1.02%). Expression of NLRP3 was lowest in combination group (58.94±1.05%), followed by folinic acid and hydroxychloroquine group, which showed equal values (60.57±1.38%), compared to control group (72.15±1.02%). Tubular injury scores were also lowest in combination group (27.07±3.16%), followed by hydroxychloroquine group (45.29±1.75%) and folinic acid group (48.38±2.49%), compared to control group (77.15±1.66%).CONCLUSION: Expression of ET-1 and NLRP3, as well as tubular injury scores, are significantly lower in all treatment groups compared to control, suggesting hydroxychloroquine and folinic acid demonstrated renoprotective effects in methanol-induced AKI, likely through modulation of ET-1 and NLRP3 pathways.KEYWORDS: methanol intoxication, acute kidney injury, hydroxychloroquine, folinic acid, endothelin-1, NLRP3 inflammasome, experimental animal models, rabbits

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