Acta Biochimica Indonesiana

Acetazolamide-mediated carbonic anhydrase inhibition suppresses human peripheral blood mononuclear cell proliferation via G1/S cell cycle arrest

2026-03-11T15:15:10+07:00

Background: Carbonic anhydrase (CA) regulates intracellular CO₂/HCO₃⁻ homeostasis and supplies carboxyl donors for the sixth step of de novo purine biosynthesis. Disruption of this step is predicted to impair nucleotide pool accumulation and arrest cell cycle progression.

Objective: This study investigated whether CA inhibition by acetazolamide suppresses T lymphocyte proliferation.

Methods: Human peripheral blood mononuclear cells (PBMCs) were stimulated with phytohemagglutinin (PHA, 1% v/v) or interleukin-2 (IL-2, 10 ng/mL). Acetazolamide was applied at 6.25–50 μM. Cell viability, DNA synthesis, and cell cycle distribution were assessed using WST-1 assay, BrdU incorporation, and propidium iodide flow cytometry, respectively.

Results: Acetazolamide reduced PBMC viability and DNA synthesis dose-dependently in both PHA- and IL-2-stimulated cultures (p < 0.05). IL-2-stimulated cells showed greater sensitivity, with significant inhibition at 12.5 μM versus 25 μM for PHA-stimulated cells. Flow cytometry revealed G1/S arrest in all treated groups: S phase decreased from 8.52% to 3.82% (PHA) and from 1.27% to 0% (IL-2) at 50 μM, with G2/M uniformly suppressed to ≤0.57%.

Conclusion: Acetazolamide suppresses PBMC proliferation through G1/S arrest, consistent with CA inhibition depleting CO₂/HCO₃⁻-dependent carboxyl donors required for de novo purine synthesis.

Turmeric extract reverses diclofenac-induced malondialdehyde elevation: comparative efficacy of two therapeutic doses

2026-02-08T11:27:32+07:00

Background: Chronic administration of non-steroidal anti-inflammatory drugs (NSAIDs) such as diclofenac sodium generates excessive reactive oxygen species (ROS), causing lipid peroxidation and elevated malondialdehyde (MDA) levels, a biomarker of oxidative tissue damage. Turmeric extract (Curcuma longa L.) contains curcuminoids with antioxidant properties that may reduce MDA formation and mitigate NSAID-induced injury.

Objectives: To determine the effect of turmeric extract on malondialdehyde (MDA) levels in white rats (Rattus norvegicus) induced with sodium diclofenac.

Methods: This laboratory experimental study employed a posttest-only control group design. Twenty-eight white rats were divided into four groups: normal control, negative control (diclofenac 10 mg/kg), treatment 1 (diclofenac + extract 100 mg/kg), and treatment 2 (diclofenac + extract 200 mg/kg). MDA levels were measured spectrophotometrically. Data were analyzed using One-Way ANOVA and post hoc Dunnett T3 test.

Results: Turmeric extract significantly reduced MDA levels (p<0.001). The 200 mg/kg dose demonstrated superior protective effects compared to 100 mg/kg, effectively preventing diclofenac-induced oxidative stress.

Conclusion: Turmeric extract demonstrates dose-dependent protective effects against diclofenac-induced oxidative stress, with 200 mg/kg achieving 18.3% MDA reduction and restoring oxidative balance. These findings support its potential as adjunct therapy for chronic NSAID users.

Rasbora borneensis fish and Vigna unguiculata legume protein supplementation restores reproductive hormone profiles in early-life protein-deficient female rats

2026-02-08T11:27:31+07:00

Background: Protein malnutrition during early development can alter the hormonal pathways essential for female reproductive maturation. It remains a significant concern in regions where inadequate protein intake contributes to stunting and long-term reproductive risks. However, evidence remains limited regarding how maternal and early-life protein deficiency affects reproductive hormone formation in offspring, and whether locally available nutrient sources can reverse these disruptions.

Objective: This study evaluated the effects of supplementation of Rasbora borneensis and Vigna unguiculata spp. cylindrica on reproductive hormones in female rat offspring subjected to protein deficiency.

Methods: A protein-deficient model was generated by feeding rats a low-protein AIN-76A diet from birth to weaning, followed by continuation of the same diet in female offspring for four weeks. The offspring were then assigned to a normal control group receiving a standard diet, a negative control group maintained on the low-protein diet, and three treatment groups receiving Seluang fish feed, cowpea cowpea feed, or a combination of both for four weeks. FSH and estrogen levels were assessed to evaluate reproductive endocrine function.

Results: All treatment diets increased FSH and estrogen levels compared with the negative control. FSH concentrations were significantly higher in all treatment groups (p = 0.012), with the combination diet yielding the greatest increase. Estrogen levels also rose substantially in the treatment groups with statistical analyses confirming significant differences (p < 0.001).

Conclusion: Seluang fish and Nagara cowpea effectively improved reproductive hormone profiles in protein-deficient female rats, highlighting their potential as relevant nutritional strategies to counter early-life protein malnutrition.

Anti-obesity potential of Eurycoma longifolia: molecular docking and experimental validation in diet-induced obese rats

2026-02-08T11:27:31+07:00

Background: Obesity is associated with hyperlipidemia and enhanced adipogenesis mediated by HMGR and GPDH enzymes. Eurycoma longifolia represents a potential natural therapeutic alternative to statins for obesity management.

Objective: To investigate the anti-obesity potential of E. longifolia compounds through HMGR and GPDH inhibition using integrated in silico and in vivo approaches.

Methods: Molecular docking was performed using AutoDock Vina to evaluate interactions between seven bioactive compounds and HMGR/GPDH proteins. Validation was confirmed by RMSD values below 2 Å. Binding affinities and dissociation constants were analyzed. In vivo study employed Wistar rats fed high-calorie, high-fat diet for 12 weeks, followed by four weeks of E. longifolia extract treatment at varying doses. Body weight and intra-abdominal fat were measured.

Results: Three compounds exhibited binding affinities equivalent to atorvastatin against HMGR (-8.4 kcal/mol). Four compounds demonstrated stronger affinity than metformin against GPDH. Extract administration significantly reduced intra-abdominal fat in a dose-dependent manner (p = 0.000).

Conclusion: E. longifolia compounds demonstrate dual inhibitory potential against HMGR and GPDH through computational modeling, with experimental validation confirming significant reduction in visceral adiposity in obese rats.

Thyroid dysfunction and serum creatinine variations: A tertiary care study from northern Kerala

2026-02-10T14:35:29+07:00

Background: Thyroid hormones regulate renal hemodynamics and glomerular filtration rate (GFR), influencing serum creatinine levels. Understanding this relationship is crucial to avoid misclassification of renal function.

Objective: To determine the association between thyroid dysfunction and serum creatinine variations in patients attending a tertiary care center.

Methods: This retrospective cross-sectional study included 84 thyroid patients (66 females, 18 males) from a tertiary care center in Calicut, Kerala. Participants were classified as hypothyroid or hyperthyroid based on thyroid function tests (TSH, FT3, FT4). Serum creatinine levels were categorized as reduced or elevated using sex-specific reference ranges. Statistical analysis employed chi-square and Fisher's exact tests.

Results: Hypothyroid patients exhibited elevated creatinine in 85-100% of cases, while hyperthyroid patients demonstrated reduced creatinine in 92-100% of cases across subgroups. Highly significant inverse associations were observed across all thyroid markers (p<0.001 for all comparisons). The strongest concordance occurred in women younger than 45 years (100% concordance), with modest attenuation in older women (≥45 years), although associations remained highly significant.

Conclusion: Thyroid dysfunction profoundly affects serum creatinine through GFR modulation. Clinicians should systematically evaluate thyroid status before diagnosing chronic kidney disease based on creatinine abnormalities to prevent misclassification.

Dose-dependent effects of Stevia rebaudiana leaf extract on malondialdehyde and catalase activity in alloxan-induced hyperglycemic rats

2026-02-08T11:27:30+07:00

Background: Diabetes mellitus-induced hyperglycemia triggers oxidative stress, characterized by elevated malondialdehyde (MDA) and impaired catalase (CAT) activity. Stevia rebaudiana, rich in steviol glycosides and polyphenols, demonstrates promising antioxidant properties, yet systematic dose-response data on oxidative stress biomarkers remain limited.

Objectives: To evaluate the dose-dependent effects of stevia leaf extract on serum MDA levels and CAT activity in alloxan-induced hyperglycemic rats.

Methods: Twenty-five male Wistar rats were allocated into normal control, diabetic control (alloxan 120 mg/kg), and three treatment groups receiving alloxan plus stevia extract at 100, 200, or 400 mg/kg body weight orally for 14 days (n=5/group). Serum MDA and CAT were measured spectrophotometrically.

Results: Diabetic control showed significantly elevated MDA (2.68±0.62 mg/dL) versus normal control (1.78±0.30 mg/dL). Stevia extract dose-dependently reduced MDA: 1.70±0.19, 1.54±0.20, and 1.38±0.09 mg/dL at 100, 200, and 400 mg/kg, respectively, representing 36.6%, 42.5%, and 48.5% reduction. The 400 mg/kg dose achieved MDA levels comparable to normal control. CAT activity showed dose-dependent restoration trend (7.92±0.76 to 8.58±0.52 mg/dL).

Conclusion: Stevia leaf extract (400 mg/kg BW) effectively reduces oxidative stress in hyperglycemic rats through significant dose-dependent MDA reduction, with potential catalase benefits requiring further investigation.

Phytochemical screening and acute oral toxicity assessment of Arenga pinnata leaf aqueous extract in Wistar rats

2026-02-09T06:36:22+07:00

Background: Arenga pinnata leaves have been traditionally used for medicinal purposes; however, their phytochemical composition and toxicity profile remain poorly characterized.

Objective: This study aimed to identify bioactive compounds in A. pinnata leaf aqueous extract and evaluate its acute oral toxicity in Wistar rats.

Methods: Qualitative phytochemical screening was performed using standard colorimetric and precipitation methods. Twenty-five male Wistar rats were randomly assigned to five groups (n=5): one control (distilled water) and four treatment groups receiving single oral doses of 200, 400, 800, and 1600 mg/kg body weight of aqueous extract. Animals were monitored for 14 days for mortality, behavioral changes, and clinical toxicity signs.

Results: Phytochemical analysis confirmed the presence of alkaloids, flavonoids, tannins, and triterpenoids. No mortality, adverse clinical signs, or pathological organ changes were observed throughout the study period. All animals exhibited normal behavior and progressive weight gain. The median lethal dose (LD₅₀) exceeded 1600 mg/kg body weight, indicating low acute oral toxicity.

Conclusion: A. pinnata leaf aqueous extract contains pharmacologically relevant bioactive compounds and demonstrates a favorable acute safety profile. These findings support traditional use while highlighting the need for sub-chronic and chronic toxicity studies.

Sustainable bromelain extraction from pineapple waste: ATPS purification, freeze-dry encapsulation, and cosmetic safety evaluation

2026-02-16T12:52:03+07:00

Background: Pineapple processing generates substantial waste; peels represent a rich source of bromelain, a proteolytic enzyme with dermatological applications.

Objective: To develop sustainable bromelain extraction from pineapple peels using aqueous two-phase system (ATPS), evaluate freeze-drying encapsulation for stabilization, and assess safety for topical cosmetic use.

Methods: Bromelain was extracted using PEG-4000/MgSO₄ ATPS and freeze-dried with encapsulating agents (gum arabic and maltodextrin). Proteolytic activity was quantified by tyrosine release and gelatin digestion assays. Purified bromelain was incorporated into gel cleanser and serum formulations (2-5%, pH 4.5-7.0) and evaluated for physicochemical properties, microbial safety, and skin compatibility through human repeat insult patch testing (HRIPT).

Results: The 10% PEG-4000/30% MgSO₄ system achieved optimal balance (37.64% yield, 2.80 U/mL activity). Freeze-drying with encapsulation produced free-flowing powder with 96.7% activity retention (372.50 GDU/g). Formulations met ASEAN microbial limits (<1,000 cfu/g) and demonstrated non-irritating properties with low hypersensitivity risk in HRIPT.

Conclusion: ATPS extraction combined with freeze-dry encapsulation produces stable, bioactive bromelain suitable for safe topical applications, demonstrating successful valorization of agricultural waste into high-value cosmetic ingredients.

Association between vitamin D receptor FokI polymorphism and hypertension in Indonesian ischemic stroke patients: a cross-sectional study

2026-03-11T14:58:13+07:00

Background: Vitamin D receptor (VDR) FokI polymorphism (rs2228570) affects VDR protein transcriptional activity and may influence hypertension risk through renin-angiotensin system modulation. However, evidence in Southeast Asian stroke populations remains limited.

Objective: To investigate the association between VDR FokI polymorphism and hypertension in Indonesian ischemic stroke patients.

Methods: This cross-sectional study enrolled 85 ischemic stroke patients (50 hypertensive, 35 normotensive) from Medan, Indonesia. VDR FokI genotypes (CC, CT, TT) were determined using PCR-RFLP. Association with hypertension was analyzed using chi-square test.

Results: Genotype distribution showed CC and CT at equal frequency (44.7% each) and TT at 10.6%, with corresponding C and T allele frequencies of 67.1% and 32.9%, respectively. Among hypertensive stroke patients, CT genotype predominated (52.0%), whereas CC genotype was most prevalent in normotensive patients (57.1%). Despite these distributional differences, no significant association was observed between VDR FokI polymorphism and hypertension (χ² = 3.732, p = 0.155).

Conclusion: VDR FokI polymorphism was not significantly associated with hypertension in Indonesian ischemic stroke patients, suggesting that genetic predisposition via this single polymorphism may be overshadowed by established stroke pathophysiology or complex gene-environment interactions in this specific clinical context.

Efficacy of liposomal amphotericin B eye drops at two concentrations in experimental aspergillus keratomycosis in rabbits

2026-03-14T09:50:06+07:00

Background: Keratomycosis, a fungal corneal infection prevalent in tropical regions, frequently leads to visual impairment and blindness. Liposomal amphotericin B (L-AmB) offers improved efficacy and reduced toxicity compared to conventional amphotericin B deoxycholate and represents a promising option for topical ophthalmic use.

Objective: To evaluate and compare the therapeutic efficacy of L-AmB eye drops at 0.15% and 0.5% concentrations in experimental Aspergillus keratomycosis in New Zealand White rabbits.

Methods: Four rabbits received midstromal corneal inoculation with a mixed suspension of Aspergillus flavus, A. fumigatus, A. niger, and A. terreus (2 x 10⁴ CFU/0.05 mL). L-AmB eye drops were applied hourly from day 8 post-inoculation until corneal cultures became negative. Antifungal susceptibility was assessed by disc diffusion. Clinical response was monitored by slit lamp examination using a modified scoring system.

Results: Negative corneal cultures were achieved in all treated eyes within 10 to 18 days (mean 15 days, L-AmB 0.15%) and 11 to 15 days (mean 14 days, L-AmB 0.5%). No toxic effects were observed. Complete corneal transparency without cicatrix was achieved in one eye treated with L-AmB 0.15%.

Conclusion: Both concentrations were effective and safe. Liposomal AmB 0.15% appears as efficacious as 0.5% for experimental Aspergillus keratomycosis. Clinical studies in humans are warranted.