Introduction:The central role of oxidative stress in neuronal injury and the progression of neurological disorders underscores the need to identify multi-target agents capable of restoring redox homeostasis. This study evaluated the antioxidant and pharmacological potential of Syzygium aromaticum ethyl acetate fraction (SEAF) using integrated phytochemical, computational, and experimental approaches. Methods:GC-MS was used to characterize the phytochemical composition of the ethyl acetate fraction of Syzygium aromaticum (SEAF). Thirteen phytometabolites with relative peak areas ≥1.0% were selected for ADME-Tox profiling and molecular docking against monoamine oxidase B (MAO-B) and 5-lipoxygenase (5-LOX). In vitro antioxidant assays, acute oral toxicity testing, and in vivo evaluation of cerebellar oxidative stress biomarkers were subsequently performed. Results:GC-MS identified eugenyl acetate (43.08%), eugenol (18.86%), and β-caryophyllene (1.64%) as the predominant metabolites. The prioritized phytometabolites exhibited favorable predicted ADME-Tox profiles and notable binding energies for MAO-B (up to -8.4 kcal/mol) and 5-LOX (up to -6.8 kcal/mol), involving key interactions with HIS367 and HIS372 in MAO-B and GLN363, TYR435, and CYS172 in 5-LOX, comparable to the standard ligands. Although ascorbic acid exhibited greater radical-scavenging and ferric-reducing potencies than SEAF in the DPPH and FRAP assays, respectively, SEAF showed higher total antioxidant activity than ascorbic acid. In vivo, SEAF administration was relatively safe up to 2000 mg/kg and significantly increased cerebellar superoxide dismutase and catalase activities while reducing malondialdehyde levels in HgCl2-treated mice, with effects comparable to vitamin E. SEAF preserved cerebellar cortex histoarchitecture and Nissl substance in Purkinje cells against mercuric chloride-evoked oxidative damage. Conclusion:Syzygium aromaticum exhibited potent antioxidant effects through combined radical scavenging, enzymatic enhancement, modulation of oxidative stress-related targets, and preservation of cerebellar histoarchitecture, supporting its potential as a candidate for further preclinical development.
Ekom Monday Etukudo,1 Ibe Michael Usman,2 Augustine Oviosun,1 Vivian Onyinye Ojiakor,1 Wusa Makena,1 Elna Owembabazi,1 Patrick Maduabuchi Aja,3 Emeka Anyanwu11Department of Anatomy, Kampala International University Western Campus, Ishaka-Bushenyi, Uganda; 2Anatomy Department, School of Medicine and Pharmacy, University of Rwanda, Butare, Rwanda; 3Department of Biochemistry, Kampala International University Western Campus, Ishaka-Bushenyi, UgandaCorrespondence: Ibe Michael Usman, Email i.m.usman@ur.ac.rwIntroduction: The central role of oxidative stress in neuronal injury and the progression of neurological disorders underscores the need to identify multi-target agents capable of restoring redox homeostasis. This study evaluated the antioxidant and pharmacological potential of Syzygium aromaticum ethyl acetate fraction (SEAF) using integrated phytochemical, computational, and experimental approaches.Methods: GC-MS was used to characterize the phytochemical composition of the ethyl acetate fraction of Syzygium aromaticum (SEAF). Thirteen phytometabolites with relative peak areas ≥ 1.0% were selected for ADME-Tox profiling and molecular docking against monoamine oxidase B (MAO-B) and 5-lipoxygenase (5-LOX). In vitro antioxidant assays, acute oral toxicity testing, and in vivo evaluation of cerebellar oxidative stress biomarkers were subsequently performed.Results: GC-MS identified eugenyl acetate (43.08%), eugenol (18.86%), and β-caryophyllene (1.64%) as the predominant metabolites. The prioritized phytometabolites exhibited favorable predicted ADME-Tox profiles and notable binding energies for MAO-B (up to − 8.4 kcal/mol) and 5-LOX (up to − 6.8 kcal/mol), involving key interactions with HIS367 and HIS372 in MAO-B and GLN363, TYR435, and CYS172 in 5-LOX, comparable to the standard ligands. Although ascorbic acid exhibited greater radical-scavenging and ferric-reducing potencies than SEAF in the DPPH and FRAP assays, respectively, SEAF showed higher total antioxidant activity than ascorbic acid. In vivo, SEAF administration was relatively safe up to 2000 mg/kg and significantly increased cerebellar superoxide dismutase and catalase activities while reducing malondialdehyde levels in HgCl2-treated mice, with effects comparable to vitamin E. SEAF preserved cerebellar cortex histoarchitecture and Nissl substance in Purkinje cells against mercuric chloride-evoked oxidative damage.Conclusion: Syzygium aromaticum exhibited potent antioxidant effects through combined radical scavenging, enzymatic enhancement, modulation of oxidative stress-related targets, and preservation of cerebellar histoarchitecture, supporting its potential as a candidate for further preclinical development.Keywords: antioxidants, cerebellum, oxidative stress, phytomedicine, Syzygium aromaticum
Background:Metabolic Associated Fatty Liver Disease (MAFLD) is a prevalent metabolic disorder with no effective approved pharmacotherapy. Andrographolide, a bioactive diterpenoid from Andrographis paniculata, has anti-inflammatory and antioxidant properties, but its mechanisms in MAFLD remain unclear. Methods:An integrated computational and experimental approach was applied. Network pharmacology was used to identify shared targets between andrographolide and MAFLD using public drug-target and disease databases. Protein-protein interaction analysis, pathway enrichment, and molecular docking with hub proteins were then performed. For experimental validation, hepatic steatosis was induced in HUH7 cells with oleic acid (1 mM, 24 h) and treated with andrographolide (14 µM). Lipid accumulation was measured by Oil Red O staining, and hub gene expression was analyzed by qRT-PCR. Results:In silico analysis identified 253 shared targets, with TNF, IL6, AKT1, TP53, IL1B, JUN, VEGFA, STAT3, CASP3, and EGFR as major hubs. Enrichment analysis highlighted lipid metabolism, and inflammation signaling pathways. Molecular docking showed strong binding to AKT1 (-7.7 kcal/mol), EGFR (-7.5 kcal/mol), and STAT3 (-7.2 kcal/mol). In vitro, andrographolide reduced oleic acid-induced lipid accumulation by 38% (p = 0.0032). qRT-PCR revealed significant downregulation of AKT1 (73.5%, p = 0.0159), EGFR (72.0%, p = 0.0137), and STAT3 (53.6%, p = 0.0220) mRNA expression in andrographolide-treated steatotic hepatocytes. These transcript-level changes are correlative observations and do not confirm direct inhibition of AKT1, EGFR, or STAT3 protein activity, which requires phosphorylation-specific validation. The mRNA findings suggest a potential association between andrographolide treatment and modulation of these signaling pathways at the transcriptional level. Conclusion:Andrographolide alleviates hepatic steatosis and is associated with transcriptional modulation of AKT1, EGFR, and STAT3 mRNA. However, protein-level validation is absent, and these findings should be considered hypothesis-generating rather than mechanistic proof of target inhibition. The results support further investigation of andrographolide as a potential multi-target therapeutic candidate for MAFLD.
Marie Louise Sezerano, Emery NiyonkuruNational Institute of Public Health, Ministry of Public Health, Bujumbura, BurundiCorrespondence: Emery Niyonkuru, National Institute of Public Health, Ministry of Public Health, Bujumbura, Burundi, Email niyoemery01@gmail.comAbstract: Moringa oleifera (M. oleifera) is a multipurpose medicinal plant with over 160 catalogued bioactive compounds and reported activity across metabolic, inflammatory, infectious, and neoplastic diseases, yet its pharmacology has remained mechanistically fragmented, limiting translational interpretation. In this narrative review, we synthesize evidence from in silico analyses, cellular systems, animal models, and human studies to evaluate whether structurally diverse phytochemicals—including glucosinolates, isothiocyanates, flavonoids, and phenolic acids—converge on interconnected redox and inflammatory regulatory networks centered on NF-κB and Nrf2/Keap1 signaling, offering a plausible systems-level explanation for their coordinated effects on immune function, oxidative stress, and metabolic homeostasis. Preclinical evidence for this convergence is extensive and consistent, with documented effects on inflammatory signaling, oxidative stress regulation, and cell fate determination relevant to hepatoprotection, cardiometabolic regulation, neuroprotection, and tumor biology. Clinical evidence, by contrast, is considerably more limited: available trials report modest and inconsistent benefits for glycemic control, cardiovascular parameters, and immune-related outcomes, constrained by heterogeneous phytochemical composition, minimal pharmacokinetic characterization, and variable extraction and formulation methods. Safety data indicate general tolerability at commonly used doses, though reproductive effects and herb–drug interactions remain incompletely defined. Collectively, current evidence positions M. oleifera as a mechanistically plausible, multi-target modulator of redox and inflammatory networks rather than a validated therapeutic agent; realizing its clinical potential will require standardized preparations, rigorous pharmacokinetic profiling, biomarker-anchored trial design, and evaluation in well-powered controlled trials.Keywords: Moringa oleifera, phytochemical bioactives, redox homeostasis, immunomodulation, multi-target pharmacology, translational therapeutics
Objective:This study aims to investigate the preventive effect of simvastatin on the subtotal nephrectomy model by analysing key mediators of inflammation and fibrosis. Methods:A total of 30 male Swiss mice were used in this study. The animals were divided into five groups (six/group): the sham group (Sham), the nephrectomy group (Neph), and three simvastatin groups (Simv) with a stepwise dose of simvastatin (5, 10, or 20 mg/kgBW). Kidney function was assessed by measuring serum creatinine, blood urea nitrogen (BUN), and proteinuria. Interstitial fibrosis was evaluated by Sirius red staining, α-SMA expression was evaluated by immunohistochemistry, and TNF-α expression was evaluated by real-time PCR. Results:Subtotal nephrectomy significantly impaired renal function, increased interstitial fibrosis, and upregulated TNF-α and α-SMA expression. Simvastatin treatment dose-dependently reduced serum creatinine and proteinuria, although the reduction in BUN was not significantly different from the Neph group. Simvastatin also significantly attenuated interstitial fibrosis (8.62±2.43 in Simv5 group, 4.15±1.67 in Simv10, and 2.62±0.63 in Simv20 groups), decreased the expression of TNF-α (0.42±0.27 in Simv5, 0.39±0.18 in Simv10, and 0.23±0.21 in Simv20) and α-SMA (3.85±0.97 in Simv5, 2.74±0.61 in Simv10, and 1.59±0.45 in Simv20 groups) in dose-dependent manner, compared with the Neph group (all p < 0.05). Path analysis demonstrated that TNF-α indirectly contributed to renal fibrosis by promoting α-SMA expression, whereas α-SMA had a significant direct effect on interstitial fibrosis (p=0.038). Conclusion:This study found that the renoprotective effect of simvastatin in a subtotal nephrectomy model may be mediated by inhibition of the TNF-α/α-SMA signalling pathway, with α-SMA acting as a key mediator linking inflammation to renal fibrosis.
Putu Nita Cahyawati,1 Erly Sintya,2 Anak Agung Sri Agung Aryastuti,1 Asri Lestarini,2 Pande Ayu Naya Kasih Permatananda11Department of Pharmacology and Pharmacy, Faculty of Medicine and Health Sciences, Universitas Warmadewa, Denpasar, Bali, Indonesia; 2Department of Biochemistry, Faculty of Medicine and Health Sciences, Universitas Warmadewa, Denpasar, Bali, IndonesiaCorrespondence: Putu Nita Cahyawati, Department of Pharmacology and Pharmacy, Faculty of Medicine and Health Sciences, Universitas Warmadewa, Jl Terompong No 24, Sumerta Kelod, Kec. Denpasar Tim, Kota, Denpasar, Bali, 80239, Email nitacahyawati@warmadewa.ac.idObjective: This study aims to investigate the preventive effect of simvastatin on the subtotal nephrectomy model by analysing key mediators of inflammation and fibrosis.Methods: A total of 30 male Swiss mice were used in this study. The animals were divided into five groups (six/group): the sham group (Sham), the nephrectomy group (Neph), and three simvastatin groups (Simv) with a stepwise dose of simvastatin (5, 10, or 20 mg/kgBW). Kidney function was assessed by measuring serum creatinine, blood urea nitrogen (BUN), and proteinuria. Interstitial fibrosis was evaluated by Sirius red staining, α-SMA expression was evaluated by immunohistochemistry, and TNF-α expression was evaluated by real-time PCR.Results: Subtotal nephrectomy significantly impaired renal function, increased interstitial fibrosis, and upregulated TNF-α and α-SMA expression. Simvastatin treatment dose-dependently reduced serum creatinine and proteinuria, although the reduction in BUN was not significantly different from the Neph group. Simvastatin also significantly attenuated interstitial fibrosis (8.62± 2.43 in Simv5 group, 4.15± 1.67 in Simv10, and 2.62± 0.63 in Simv20 groups), decreased the expression of TNF-α (0.42± 0.27 in Simv5, 0.39± 0.18 in Simv10, and 0.23± 0.21 in Simv20) and α-SMA (3.85± 0.97 in Simv5, 2.74± 0.61 in Simv10, and 1.59± 0.45 in Simv20 groups) in dose-dependent manner, compared with the Neph group (all p < 0.05). Path analysis demonstrated that TNF-α indirectly contributed to renal fibrosis by promoting α-SMA expression, whereas α-SMA had a significant direct effect on interstitial fibrosis (p=0.038).Conclusion: This study found that the renoprotective effect of simvastatin in a subtotal nephrectomy model may be mediated by inhibition of the TNF-α/α-SMA signalling pathway, with α-SMA acting as a key mediator linking inflammation to renal fibrosis.Keywords: anti-inflammatory, antifibrotic, kidney disease, simvastatin, TNF-α, α-SMA
Genetic factors, dietary habits, and lifestyle may influence individual responses to supplementation, including the bergamot flavanones naringin and naringenin, which are implicated in lipid modulation. The aim of this double-blind, placebo-controlled clinical trial (RCT) is to assess the effects of the supplementation for four months of a standardised bergamot extract at the dosage of 375 mg/day, providing 150 mg/day of flavonoids, on lipid parameters of 108 East Asian adults with LDL-Cholesterol (LDL-C) 100-159 mg/day, since to date, the RCTs have been conducted on Caucasian subjects only. After 4 months, total Cholesterol and LDL-C decreased (-7.3% and -7.0%) with no clinically relevant safety changes regarding hepatic/renal biochemistry, weight, and blood pressure.
Background:Myrothamnus flabellifolius (M. flabellifolius) herb is widely used across Southern Africa, often with poorly defined or unrestricted dosing and limited safety considerations across its diverse ethnomedicinal applications. Objective:This study investigated the effects of M. flabellifolius crude aqueous extract on rat liver and kidneys. Methods:M. flabellifolius crude aqueous extract was screened for its bioactive components. Safety was assessed using single and 28-day repeated oral dose toxicity studies. In the former, male and female Sprague-Dawley rats (n = 12 per group; 6 per sex) received M. flabellifolius stem and leaf crude aqueous extract at doses of 0, 500, 1000, 2000, 5000 and 10000 mg/kg and were observed for 14 days for clinical signs, body weight changes, and mortality. Based on the absence of clinical signs and lethality, the same dose range was applied in the 28-day repeated oral dose toxicity study in which female rats (n = 6 per group) received treatment. At the end of the study, blood samples were collected for biochemical analysis, while liver and kidney samples were harvested for relative organ weight and histopathological examination. Results:Phytochemical screening confirmed the presence of tannins, flavonoids, coumarins and terpenoids in the extract. Treatment in both single and 28-day repeated oral dose toxicity studies showed no clinical signs or mortality; body/relative organ weights changes were non-significant (p > 0.05). The oral no-observed-adverse-effect level (NOAEL) was 10000 mg/kg in both sexes. Biochemical indices showed non-significant changes (p > 0.05), whereas liver and kidney histoarchitecture remained unremarkable. Conclusion:M. flabellifolius crude aqueous extracts exhibited a wide safety margin with no detectable hepatorenal toxicity. Its confirmed bioactive phytochemicals valorizes ethnomedicinal use.
Male infertility contributes to nearly half of all infertility cases worldwide and is closely linked to oxidative stress, inflammation, endocrine imbalance, and environmental toxicant exposure, all of which impair spermatogenesis and sperm function. Interest in naturally derived bioactive compounds has increased due to their potential multitarget therapeutic effects and relatively favorable safety profiles. Acetyl-eugenol, a structural derivative of eugenol found in Syzygium aromaticum and other aromatic plants, has demonstrated antioxidant, anti-inflammatory, and cytoprotective activities in experimental studies. This review critically evaluates the potential role of acetyl-eugenol in male reproductive health, focusing on its pharmacological properties and proposed mechanisms of action. Available preclinical evidence suggests that acetyl-eugenol may reduce oxidative stress by scavenging reactive oxygen species, enhancing endogenous antioxidant defenses, and modulating inflammatory pathways such as NF-κB signaling. It may also contribute to the preservation of testicular structure and function, including protection of Sertoli and Leydig cells. However, current evidence remains limited and is derived predominantly from in vitro and animal studies, with very limited direct evidence in humans. Many proposed reproductive benefits are inferred from studies involving eugenol or related compounds rather than acetyl-eugenol specifically. In addition, conflicting findings regarding dose-dependent toxicity, insufficient pharmacokinetic data, and the absence of standardized clinical studies limit definitive conclusions regarding efficacy and safety. Overall, acetyl-eugenol represents a promising candidate for mitigating oxidative stress-associated male reproductive dysfunction, but its therapeutic potential remains preliminary. Further well-designed experimental and clinical studies are necessary to clarify its mechanisms, safety profile, and translational relevance in male infertility management.
Background:Type 2 diabetes mellitus (T2DM) is characterized by progressive β-cell dysfunction driven in part by palmitic acid (PA)-induced lipotoxicity. Cyanidin-3-O-glucoside (C3G) possesses antioxidant properties, but its temporal and mechanistic effects at higher concentrations remain poorly defined. Objective:This study aimed to investigate the continuous effects of high‑concentration of C3G on PA‑induced β‑cell apoptosis, with emphasis on time‑dependent efficacy and underlying antioxidant mechanisms. Methods:MIN6 cells were treated with PA (500 µM) in the presence or absence of C3G (500 µM). Cell viability was assessed by MTT assay, apoptosis by flow cytometry and cell death assay, oxidative stress by DCFDA and DPPH assays, and ER-stress related C/EBP homologous protein (CHOP) and apoptotic cleaved caspase-3 expression by Western blot analysis. Results:PA (500 µM) significantly reduced MIN6 cell viability, whereas continuous exposure to C3G (500 µM) restored viability, corresponding to a 40.9% increase (p < 0.001) compared with PA treatment alone. Flow cytometry showed that PA reduced live cells from 78.6% to 41.0% and increased early and late apoptotic populations to 27.7% and 28.2%, respectively, whereas C3G co-treatment reduced apoptosis to 17.7%. Consistently, cell death assays confirmed that C3G lowered apoptotic cells from 30.7% to 5.0% (p < 0.001 vs PA-treated groups) and restored viability to 93.1%, comparable to vehicle controls. Reactive oxygen species (ROS) assays, including DCFDA and DPPH, further demonstrated that C3G reduced PA‑induced ROS by 26.5% (p < 0.01 vs PA alone), consistent with free radical‑scavenging activity. Western blot analyses revealed that PA markedly upregulated CHOP and cleaved caspase‑3, which was suppressed by co‑treatment with C3G, confirming the attenuation by C3G of ER stress‑mediated apoptotic signaling. These findings imply that C3G rapidly counteracts oxidative stress-induced apoptosis. Conclusion:High-concentration C3G mitigates PA-induced oxidative stress and ER stress-associated apoptosis in β-cells, supporting its potential as a therapeutic agent for T2DM.
Introduction:Bisphenol A (BPA), a well-known endocrine disruptor, is newly emerging as causing male infertility by endocrine disturbances, induction of oxidative stress and testicular toxicity. This study investigates the protective role of hesperidin against BPA-induced testicular toxicity in male Wistar rats. Methodology:The rats were divided into six groups (n=7): control (normal saline); BPA-only (50 mg/kg); hesperidin-only at 50 mg/kg and 100 mg/kg; and two pre-treated groups receiving hesperidin (50 and 100 mg/kg) followed by BPA (50 mg/kg). All treatments were administered orally for 8 weeks. Post-treatment analyses included testicular weight, volume, and diameter measurements; serum testosterone, FSH, and LH levels; oxidative stress markers (MDA, SOD, CAT, GPx, and GSH); and histological and morphometric evaluation of the testis. Result and Discussion:BPA exposure significantly reduced testicular dimensions, decreased serum testosterone, FSH, and LH. Histopathological examination revealed degeneration of seminiferous tubules, germ-cell depletion, reduced counts of spermatogonia, spermatocytes, spermatids, Sertoli and Leydig cells. BPA significantly induced oxidative stress, evidenced by increased MDA and decreased SOD, CAT, GPx, and GSH. Hesperidin significantly mitigated these effects by restoring testicular morphology, hormonal profiles, enhancing oxidative stress markers, improving histology and cell populations. Conclusion:Hesperidin demonstrates protective properties against BPA-induced testicular and endocrine toxicity in rats, likely as a result of its antioxidant properties.
Background:Tribulus terrestris (T. terrestris) is traditionally used in the management of urinary disorders; however, evidence regarding its safety upon repeated administration and its antiurolithiatic efficacy remains limited. This study aimed to evaluate both the sub-acute toxicity and antiurolithiatic potential of the ethanol extract of T. terrestris (EETT) in experimental rats. Methods:Sub-acute toxicity was assessed in Wistar rats (n = 30; 5 males and 5 females per group) following 28-day repeated oral administration of EETT (500 and 1000 mg/kg). Antiurolithiatic activity was evaluated in an ethylene glycol/ammonium chloride-induced nephrolithiasis model (n = 36; 6 rats per group), where EETT (150, 300, and 450 mg/kg, intraperitoneally) was administered and compared with potassium citrate. Hematological, biochemical, and histopathological parameters were analyzed. Results:EETT exhibited no mortality or treatment-related toxicity, with no significant alterations in body weight, relative organ weights, hematological indices, or serum biochemical parameters. Histological examination confirmed normal tissue architecture in treated animals. In the nephrolithiasis model, EETT significantly (P < 0.05) reduced elevated serum creatinine, urea, calcium, and potassium levels in a dose-dependent manner. Histopathological findings revealed decreased calcium oxalate crystal deposition and preservation of renal structure, particularly at higher doses, with effects comparable to potassium citrate. Conclusion:The findings demonstrate that EETT possesses a favorable safety profile alongside significant antiurolithiatic activity. This dual evidence supports its potential as a therapeutic candidate for nephrolithiasis and warrants further pharmacological and clinical investigation.
Plants of the genus Strobilanthes are one of the most widespread vegetation in Southeast Asia. More than 100 metabolites, including alkaloids, fatty acids and derivatives, flavonoids and flavonoid glycosides, phenolic acids, sterols, and terpenoids, have been identified from these plants. The scope of this review, which covers ethnobotanical, phytochemical, and preclinical aspects of Strobilanthes crispus and Strobilanthes cusia (Acanthaceae), is limited to articles published between 2015 and 2025. It confirms that these plants are still being investigated globally, with the most reported in numerous in vitro and in vivo studies on the leaves. Considering the noteworthy findings of the in vitro and in vivo studies, which mainly point to S. crispus, this plant may be established as a plant-based antimicrobial, anti-inflammatory, anticancer, or hypoglycemic agent, which may be attributed to its indolo-quinazoline alkaloid and flavonoids. Despite promising pharmacological evidence, there are limited human studies during the selected publication period. However, several articles have described ethnopharmacological surveys of medicinal plants in China, Malaysia, and Thailand, which documented the folkloric use of these two plants. It should be taken to notice that the lack of human studies requires further clinical trials to validate pharmacological activity, efficacy, and safety, and confirm its potential as a therapeutic agent.
Introduction:Schizophrenia involves dopaminergic dysregulation, oxidative stress, and glial activation within motor-cognitive circuits. Mangiferin, a polyphenolic C-glucoside from Mangifera indica, exerts antioxidant and anti-inflammatory effects partly via modulation of nuclear factor erythroid 2-related factor 2 (Nrf2) signaling. This study evaluated whether mangiferin attenuates ketamine-induced behavioral and neurobiological alterations along the basal ganglia-substantia nigra-cerebellar axis in rats. Methods:Male Wistar rats were assigned to seven groups (n = 6) and received vehicle, ketamine (50 mg/kg/day, i.p. 7 days), mangiferin (25-75 mg/kg, p.o. 14 days), ketamine plus mangiferin (25, 50, 75 mg/kg), or ketamine plus risperidone (2 mg/kg, p.o). Y-maze and open-field tests were conducted at baseline, after ketamine, and after treatment. Striatum, substantia nigra, and cerebellum were analyzed for dopamine (HPLC), oxidative stress markers, inflammatory mediators, and immunohistochemistry for GFAP and Nrf2. Results:Ketamine produced behavioral alterations characterized by reduced exploratory activity, hyperlocomotion, and anxiety-like behavior, alongside elevated dopamine, reduced antioxidant enzyme activities, increased lipid peroxidation and pro-inflammatory mediators, enhanced GFAP immunoreactivity, and decreased Nrf2 immunoreactivity. Mangiferin, particularly at 50-75 mg/kg, increased Y-maze arm entries toward control values (indicating improved locomotor activity), restored antioxidant defenses, reduced oxidative and inflammatory indices toward control levels, reduced astrocytosis, and increased Nrf2 immunoreactivity. Risperidone improved behavior and neuroinflammatory indices but showed less consistent normalization of redox markers and Nrf2 compared with high-dose mangiferin. Discussion:These findings indicate that mangiferin attenuates ketamine-induced behavioral, oxidative, inflammatory, and glial alterations in motor-cognitive circuits and are consistent with modulation of redox-glial interactions, including Nrf2-associated antioxidant signaling. Collectively, the results support further evaluation of mangiferin and related Nrf2-modulating natural products as adjunctive strategies targeting redox-glial dysfunction in schizophrenia.
Introduction:Ischemia/reperfusion (I/R) injury is a critical complication associated with pancreatic surgery, and transplantation, which frequently develops into acute pancreatitis due to increased oxidative stress and inflammatory cascades. Aim:This study aimed to assess the protective effects of vulgarin (VLG) and epivulgarin (EPV) against pancreatic I/R- injury in rats. Methods:Rats were given oral dosages of 10 or 20 mg/kg of VLG or EPV for two days prior to I/R and 24 h after reperfusion. Pancreatic I/R was induced by occluding the pancreatic blood supply for 60 minutes followed by reperfusion. Key biochemical markers including serum amylase, lipase, tumor necrosis factor-alpha (TNF-α), interleukin-1β (IL-1β), malondialdehyde (MDA), and glutathione peroxidase (GPx) were measured. Additionally, pancreatic tissue expression of high mobility group box 1 (HMGB1) and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) was assessed via immunohistochemistry. Results:Pancreatic I/R significantly elevated serum levels of amylase (4.5-fold) and lipase (5.8-fold), oxidative stress marker; MDA (8.5-fold), as well as pro-inflammatory cytokines; TNF-α (10.4-fold) and IL-1β (5.6-fold) compared to controls (p≤0.01). The antioxidant GPx activity was suppressed by 84% (p≤0.05). Treatment with VLG and EPV dose-dependently attenuated these changes, with the highest dose of EPV reducing serum amylase by 72% and MDA by 76% compared to untreated I/R rats (p≤0.01). Immunohistochemical analysis revealed marked downregulation of HMGB1 and NF-κB expression in VLG and EPV-treated groups. High-dose therapies showed near normalization of numerous biochemical and molecular parameters. Conclusion:Vulgarin and epivulgarin exhibited significant protective effects against pancreatic I/R injury by reducing oxidative stress and suppressing HMGB1/NF-κB-mediated inflammatory signaling. These findings suggest the therapeutic potential of VLG and EPV in managing acute pancreatitis and related pancreatic injuries.
Background:Liver fibrosis represents a serious health challenge and is the outcome of chronic liver diseases like cirrhosis and hepatitis. This study was designed to evaluate the anti-fibrotic and anti-dyslipidemic effects of Livogrit (a tri-herbal formulation) on Thioacetamide (TAA)-induced rat model of fibrosis; and its mutagenic potential in Ames test. Methods:The study employed TAA-induced Sprague-Dawley rat to evaluate Livogrit's anti-fibrotic potentials, as well as associated dyslipidemia. Quantification of phytometabolites present in Livogrit was conducted using UHPLC-DAD analysis. The pharmacological effects of Livogrit were assessed by measuring hepatic enzyme markers AST, ALT, and ALP; serum lipid profile markers TG, TC, HDL, and LDL; anti-oxidative enzymes SOD and catalase. In addition, histopathological changes in hepatic tissue were assessed. Changes in body weight, relative liver weight, hydroxyproline, and collagen levels were also evaluated. Silymarin served as the experimental reference standard. Finally, Livogrit was also tested for its mutagenic potential through Ames assay. Results:UHPLC-DAD analysis of Livogrit revealed presence of several bioactive metabolites. Livogrit effectively attenuated TAA-induced hepatotoxicity and fibrosis. Treatment with Livogrit reduced elevated ALT, AST, ALP, TG, TC, LDL, nitrite, hydroxyproline, and collagen levels while improving HDL, SOD, and catalase levels. Livogrit also regulated LDL/HDL and TC/HDL ratios. Livogrit treatment normalized the detrimental effects of TAA on the liver histo-architecture, in terms of inflammatory and fibrotic changes. Ames test also confirmed that Livogrit was non-mutagenic at highest tested concentration, with or without metabolic (S9) activation. Conclusion:Livogrit demonstrated preclinical potential for the effective management of hepatic fibrosis and dyslipidemia, in a non-mutagenic manner. This study paves a way for detailed non-clinical safety experiments and clinical investigations of Livogrit in patients with hepatic fibrosis under controlled conditions.
Acharya Balkrishna,1– 3 Sunil Shukla,1 Sandeep Sinha,1 Himanshu Jangid,1 Savita Lochab,1 Anurag Varshney1,2 1Drug Discovery and Development Division, Patanjali Research Foundation, Haridwar, Uttarakhand, 249405, India; 2Department of Allied and Applied Sciences, University of Patanjali, Haridwar, Uttarakhand, 249405, India; 3Patanjali Yog Peeth (UK) Trust, Glasgow, G41 1AU, UKCorrespondence: Anurag Varshney, Drug Discovery and Development Division, Patanjali Research Foundation, NH-58, Near Bahadrabad, Haridwar, Uttarakhand, 249405, India, Tel +91 1334-244107 Ext. 7458, Fax +91 1334 244805, Email anurag@prft.co.in; anurag@patanjali.res.inBackground: Liver fibrosis represents a serious health challenge and is the outcome of chronic liver diseases like cirrhosis and hepatitis. This study was designed to evaluate the anti-fibrotic and anti-dyslipidemic effects of Livogrit (a tri-herbal formulation) on Thioacetamide (TAA)-induced rat model of fibrosis; and its mutagenic potential in Ames test.Methods: The study employed TAA-induced Sprague-Dawley rat to evaluate Livogrit’s anti-fibrotic potentials, as well as associated dyslipidemia. Quantification of phytometabolites present in Livogrit was conducted using UHPLC-DAD analysis. The pharmacological effects of Livogrit were assessed by measuring hepatic enzyme markers AST, ALT, and ALP; serum lipid profile markers TG, TC, HDL, and LDL; anti-oxidative enzymes SOD and catalase. In addition, histopathological changes in hepatic tissue were assessed. Changes in body weight, relative liver weight, hydroxyproline, and collagen levels were also evaluated. Silymarin served as the experimental reference standard. Finally, Livogrit was also tested for its mutagenic potential through Ames assay.Results: UHPLC-DAD analysis of Livogrit revealed presence of several bioactive metabolites. Livogrit effectively attenuated TAA-induced hepatotoxicity and fibrosis. Treatment with Livogrit reduced elevated ALT, AST, ALP, TG, TC, LDL, nitrite, hydroxyproline, and collagen levels while improving HDL, SOD, and catalase levels. Livogrit also regulated LDL/HDL and TC/HDL ratios. Livogrit treatment normalized the detrimental effects of TAA on the liver histo-architecture, in terms of inflammatory and fibrotic changes. Ames test also confirmed that Livogrit was non-mutagenic at highest tested concentration, with or without metabolic (S9) activation.Conclusion: Livogrit demonstrated preclinical potential for the effective management of hepatic fibrosis and dyslipidemia, in a non-mutagenic manner. This study paves a way for detailed non-clinical safety experiments and clinical investigations of Livogrit in patients with hepatic fibrosis under controlled conditions. Keywords: liver fibrosis, dyslipidemia, Livogrit, thioacetamide, histopathology, Ames assay, UHPLC
Purpose:Carica papaya is used alone or with other herbs in traditional medicine to treat infectious diseases; nonetheless, comprehensive scientific validation of this claim is limited. We conducted this preliminary study to explore its immunomodulatory potential through cytokine modulation. Methods:An aqueous ethanolic crude extract of plant leaves was evaluated for phytochemical groups, total flavonoid, and phenolic content using aluminum chloride and Folin-Ciocalteu's methods. The crude extract and its derived fractions were compared with thin-layer chromatography, and antioxidant properties were assessed using the 2,2'-diphenyl-1-picrylhydrazyl radical scavenging assay and the Ferric Ion Reducing Antioxidant Power assay. Additionally, the effects of the extracts on cell proliferation, cytokine, and nitric oxide release by mouse peritoneal macrophages and bone marrow multipotent cells in culture were evaluated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, the Enzyme-Linked Immunosorbent Assay (ELISA), and the Griess reagent. The cells were collected from Swiss strain mice (12 mice). Results:Carica papaya leaf crude extract contained alkaloids, tannins, flavonoids, anthraquinones, and terpenoids. The crude extract's total phenolic and flavonoid contents were 14.07±5.76 gallic acid equivalents and 22.58±3.05 quercetin equivalents per gram of extract. The crude extract showed more potent antioxidant activity. Crude extract and fractions stimulated the proliferation of peritoneal macrophages at 1-10μg/mL and bone marrow multipotent cells at 0.01-0.3μg/mL in a concentration-dependent manner. Interleukin-6 and tumor necrosis factor-α production were significantly increased, while interleukin-1β production was inhibited in treated peritoneal macrophages. Treated bone marrow cells increased granulocyte-macrophage colony-stimulating factor, stem cell factor, and thrombopoietin secretion and decreased interleukin-3 production. Nitric oxide production was 10-20 times higher in bone marrow compared to peritoneal macrophage cells. Conclusion:Carica papaya leaf crude extract and fractions exhibit immunomodulatory effects by promoting cytokine release and peritoneal macrophage and bone marrow cell proliferation, explaining the use of C. papaya in infectious diseases in traditional medicine. Further exploration is needed for applications in immune-related diseases.
Purpose:Triple-negative breast cancer (TNBC) is an aggressive breast cancer subtype with limited targeted treatment options. Doxorubicin remains a cornerstone of TNBC treatment; however, its molecular effects beyond canonical cytotoxic mechanisms are not fully characterized. This study aimed to explore NF-κB- and HIF-1α-related transcriptional responses associated with doxorubicin treatment in TNBC cells using an integrative computational and experimental approach, in line with global cancer research priorities supporting Sustainable Development Goal (SDG) 3: Good Health and Well-Being. Methods:Network pharmacology analysis, molecular docking, and molecular dynamics simulations were employed to explore potential pathway-level associations of doxorubicin with NF-κB and HIF-1α-related signaling. In vitro validation was performed using MTT cytotoxicity assays in MDA-MB-231 cells cultured in DMEM and RPMI-1640 media. Half-maximal inhibitory concentrations (IC5 0) were determined using four-parameter logistic regression. Transcriptional responses of NF-κB and HIF-1α were evaluated by RT-qPCR under normoxic conditions. Results:Computational analyses suggested potential associations between doxorubicin and components of NF-κB and HIF-1α-related signaling pathways. In vitro assays demonstrated concentration-dependent cytotoxicity, with IC5 0 values of 2.34 µM (95% CI: 2.11-2.74) in DMEM and 1.07 µM (95% CI: 0.92-1.32 µM) in RPMI-1640. RT-qPCR analysis revealed downregulation of NF-κB and HIF-1α mRNA expression following doxorubicin treatment. These findings indicate transcriptional modulation associated with doxorubicin exposure, without establishing functional pathway inhibition. Conclusion:This study provides transcriptional-level evidence suggesting the involvement of NF-κB- and HIF-1α-related pathways in the cellular response of TNBC cells to doxorubicin treatment. By integrating computational predictions with early experimental validation, the findings generate biologically plausible hypotheses for further mechanistic and functional investigations, contributing to foundational cancer research efforts aligned with SDG 3 (Good Health and Well-Being).
Objective:Considering the historical use of three plant species Punica granatum, Quercus infectoria, and Achillea millefolium in traditional medicine for hemostatic purposes, along with the variation observed in some experimental research, the aim of this study was to comparatively evaluate the effect of methanol extracts derived from these three medicinal plants on blood clotting time in vitro. Methods:Methanolic (70%) extracts of P. granatum flower, Q. infectoria galls and A. millefolium aerial parts were prepared by maceration method. The total tannin and phenolic content were determined using a colorimetric method. The cytotoxic activity of extracts on NIH/3T3 cell lines was evaluated by the colorimetric MTT assay. Non-toxic concentrations were tested in vitro on blood coagulation profiles including prothrombin time (PT) and activated partial thromboplastin time (aPTT) of healthy human volunteers. Results:Q. infectoria had the highest phenolic content at 287.18 mg gal/g, followed by P. granatum and A. millefolium. P. granatum exhibited the highest tannin content (119.21 mg catechin/g DW), followed by Q. infectoria (109.30 mg catechin/g DW) and A. millefolium (54.35mg catechin/g DW). The highest total flavonoid content was found in P. granatum, A. millefolium, and Q. infectoria with 98, 85.5, 5.22 mg Rutin/g DW, respectively. Non-toxic concentrations were selected to perform coagulation experiments. Based on the plant-derived coagulation assays, pomegranate extract notably reduced the PT to 11.57 ± 0.37 s compared with the control value of 14.47 ± 0.41 s. Conversely, gall oak extract markedly increased the coagulation time to 17.07 ± 1.07 s. Similarly, pomegranate significantly decreased aPTT to 33.83 ± 0.79 s relative to the control (36.30 ± 2.04 s), whereas gall oak considerably prolonged it to 50.00 ± 2.04 s. Conclusion:Methanolic extracts of P. granatum show promise as procoagulants, while Q. infectoria acts as an anticoagulant, and A. millefolium remains neutral in this context.
