Search Results (157)

Ginsenoside accumulation is tissue- and germplasm-specific, yet its transcriptional regulation remains unclear. We combined HPLC, PacBio full-length transcriptome, and Illumina RNA-seq to compare 4-year cultivated ginseng with 26-year wild-simulated ginseng. After clustering and error correction, we obtained 265,373 (CG) and 210,343 (WSG) non-redundant full-length transcript isoforms. Total saponins ranked leaves > roots > stems. In leaves, PPT type (Re and Rg1) predominated and was significantly higher in CG than WSG; in roots, PPD type dominated, and all monomer saponins in WSG were significantly higher. Transcriptome analysis revealed widespread alternative splicing (mainly retained introns), with key genes like CYP716A47 showing differential expression and splicing. WGCNA identified a yellow module positively correlated with PPD-type ginsenosides, enriched in terpenoid and CYP/UGT pathways and containing CYP716A47 and multiple UGT candidates. qRT-PCR validated 12 selected genes (6 known, 6 novel). Our findings reveal multilevel transcriptional and post-transcriptional regulation associated with the distinct production backgrounds of cultivated and wild-simulated ginseng and provide candidate targets for future functional studies and metabolic engineering. Full article

Background: The emergence of antifungal resistance and the limited number of clinically available antifungal drug classes necessitate the development of new agents with improved efficacy and safety. We investigated how linker architecture influences the antifungal activity and lead properties of hybrid triazole-thiazolidine derivatives. Methods: A focused library of triazole-thiazolidine hybrids incorporating alkylamine, amide, cyclic amine, 2-hydroxypropyl, and thiazepane linkers was synthesized and characterized. Antifungal activity was evaluated against reference strains and clinical isolates of Candida spp., Aspergillus fumigatus, dermatophytes, and Cryptococcus neoformans. Structure–activity relationships were analyzed by molecular docking. Selected compounds were further assessed by SCRAPPY profiling, fluorescence microscopy, mammalian-cell cytotoxicity assays, acute oral toxicity studies, and evaluation of microsomal stability and interactions with human CYP450 isoforms. Results: Linker architecture strongly influenced antifungal potency. Amide- and cyclic amine-containing hybrids were generally the most active, whereas simple alkylamide derivatives showed narrower activity profiles. Compound 28 emerged as the most promising lead, exhibiting sub-microgram MIC values against several Candida isolates, particularly C. parapsilosis, and retaining measurable activity against an azole-resistant C. albicans strain. Docking generated putative CYP51-binding models, while SCRAPPY profiling and fluorescence microscopy revealed an azole-like cellular response consistent with perturbation of sterol-associated homeostasis. Compound 28 was tolerated at 300 mg/kg in an acute oral study but showed concentration- and time-dependent cytotoxicity and rapid CYP3A4-mediated microsomal metabolism. Conclusions: Systematic variation of linker architecture identified compound 28 as a promising exploratory antifungal lead. Further optimization should focus on improving metabolic stability and cytotoxicity, together with direct target validation, pharmacokinetic characterization, and in vivo efficacy studies. Full article

Peroxisome proliferator-activated receptors (PPARs) are ligand-activated nuclear transcription factors comprising three isoforms—PPARα, PPARγ, and PPARβ/δ—that regulate hepatic lipid metabolism, glucose homeostasis, inflammation, bile acid synthesis, and fibrogenesis. Because liver diseases involve overlapping metabolic, inflammatory, cholestatic, and fibrotic pathways, PPAR agonists have emerged as a versatile therapeutic class across a spectrum of hepatic conditions. PPARα agonists (e.g., fenofibrate) promote fatty acid β-oxidation and suppress de novo lipogenesis; PPARγ agonists (e.g., pioglitazone) improve insulin sensitivity and exert anti-inflammatory and antifibrotic effects; and PPARδ agonists (e.g., seladelpar) regulate bile acid and cholesterol metabolism. Dual agonists (elafibranor [PPARα/δ] and saroglitazar [PPARα/γ]) and pan-PPAR agonists (lanifibranor [PPARα/γ/δ] and bezafibrate) aim to simultaneously address multiple pathogenic mechanisms. In primary biliary cholangitis (PBC), elafibranor and seladelpar received accelerated FDA approval in 2024 based on phase 3 trials (ELATIVE and RESPONSE, respectively), demonstrating significant biochemical response rates of 51% and 62% versus 4% and 20% with the placebo. Long-term open-label extension data from the ELATIVE trial have demonstrated sustained improvements in cholestatic biomarkers and stabilization of fibrosis markers over three years, with durable benefits on fatigue and pruritus. The ASSURE open-label study has confirmed the durability of seladelpar’s effects on biochemical response and pruritus through up to two years of treatment. Saroglitazar, a dual PPARα/γ agonist, has shown positive topline phase 3 results in the EPICS-III trial and received an FDA priority review designation. Bezafibrate has shown a survival benefit in large retrospective analyses and is used as a second-line therapy in Europe and Japan; notably, bezafibrate functions as a dual PPAR/pregnane X receptor (PXR) agonist, inducing CYP3A4 and efflux transporters that contribute to bile acid detoxification. In metabolic dysfunction-associated steatotic liver disease (MASLD)/metabolic dysfunction-associated steatohepatitis (MASH), pioglitazone remains the most extensively studied PPAR agonist, with meta-analytic evidence supporting MASH resolution and fibrosis reduction regardless of diabetes status. Lanifibranor demonstrated histological improvement in the phase 2b NATIVE trial and is currently in phase 3 development (NATiV3). PPAR agonists have also demonstrated therapeutic effects on liver fibrosis inhibition through direct modulation of hepatic stellate cell activation and suppression of fibrogenic signaling. This narrative review synthesizes the molecular pharmacology of PPAR isoforms; the available clinical and preclinical evidence for mono-, dual-, and pan-PPAR agonists; and their therapeutic applications across MASLD/MASH, alcohol-associated liver disease (ALD), PBC, primary sclerosing cholangitis (PSC), intestinal failure-associated liver disease (IFALD), and advanced chronic liver disease (ACLD). The evolution from single-isoform to multi-isoform PPAR agonism reflects the recognition that overlapping pathogenic mechanisms in liver diseases may require broader receptor coverage for optimal therapeutic efficacy. Full article

The seeds of Polygonatum sibiricum exhibit dormancy, which poses a major challenge for its cultivation. To elucidate the regulatory mechanisms underlying dormancy release, we performed integrated transcriptomic and metabolomic analyses on seeds at 0, 5, 10, and 15 d after seed imbibition. Physiological assays revealed progressive declines in abscisic acid (ABA) and starch levels, alongside increases in gibberellin (GA) and soluble sugar contents, reflecting the metabolic changes accompanying the transition from dormancy to germination. Transcriptomic analysis identified 11,520 expressed genes, with 6753, 7775 and 9387 differentially expressed genes (DEGs) at T5, T10, and T15, respectively. KEGG enrichment highlighted starch and sucrose metabolism and plant hormone signal transduction as key pathways. Notably, the GA biosynthesis gene GA3ox was markedly upregulated, while the DELLA repressor (Isoform0012761) showed sustained downregulation, suggesting relieved GA signaling. In the ABA pathway, CYP707A catabolic genes exhibited biphasic expression, and ABA signaling components (PYL, PP2C, SnRK2) showed stage-specific remodeling. A total of 316, 412, and 479 differentially expressed transcription factors were identified across stages, with the GRAS family being the largest. Co-expression network analysis revealed 19 transcription factors integrating starch/sucrose metabolism with ABA and GA signaling, most of which were downregulated, except one C2H2 member showing sustained upregulation. These findings demonstrate that dormancy release in P. sibiricum is governed by coordinated hormonal reprogramming, metabolic mobilization, and transcription factor-mediated regulation, providing a theoretical foundation for improving seed germination in this medicinal plant. Full article

Background: Multiple symmetric lipomatosis (MSL) is an orphan disease characterized by pathological expansion of lipomatous tissue (LT) in the subcutaneous adipose tissue (SAT) areas, causing severe deformities and compression of vital organs. To date, the only treatment options available are either lifestyle modification or surgical excision/liposuction. Since sex hormones play an important role in adipose tissue (AT) expansion, they may represent an opportunity for pharmacological treatment. Methods: LT and control SAT were collected from eight MSL patients. Both tissues and derived stromal vascular fraction (SVF) were compared in terms of aromatase (hCYP19A1) and 5-α-reductase isoform (hSRD5A1, -2 and -3) mRNA expression. Primary cultures obtained from one MSL patient were exposed to an aromatase inhibitor, 5-α-reductase inhibitor and DL-α-Lipoic Acid and assayed for cell viability, proliferation and adipogenic differentiation. Results: hCYP19A1, hSRD5A1 and hSRD5A3 were expressed in both MSL-affected and MSL-spared tissues at different levels. Cell viability and proliferation of LT-derived SVF cells were slightly increased by treatment with DL-α-Lipoic Acid and the aromatase inhibitor. Adipogenesis was slightly reduced by Finasteride and strongly reduced by Exemestane (more in LT- than in SAT- derived primary cultures). Conclusions: These preliminary results pave the way for the use of aromatase and 5-α-reductase inhibition as a potential pharmacological approach to tackling dysfunctional adipogenesis in MSL. Therefore, the reported study should be interpreted as exploratory and further experiments are recommended in a larger number of patients to validate these promising results. Full article

Pulmonaria rubra (Boraginaceae) is a widely distributed plant in Bulgaria, yet its phytochemical profile and therapeutic potential have remained unexplored. P. rubra methanol extract (PRE) was evaluated through phytochemical profiling and in vitro neuroprotective and antioxidant assays. Rat brain synaptosomes, mitochondria and microsomes were treated with PRE alone, and in combination with 6-hydroxydopamine and tert-butyl hydroperoxide as toxic agents. The extract exhibited concentration-dependent protective effects in all subcellular models. Additionally, it was tested on hMAOA/B and different isoforms of CYP450 enzymes, but it did not show any activity in the tested conditions. In the UHPLC-HRESIMS analysis, 26 secondary metabolites were identified, mainly hydroxycinnamic acids and caffeoyl oligomers, flavonoids, a lignan (globoidnan A), and the terpenoid glycoside roseoside. Seven compounds were identified via UHPLC-UV method using reference compounds: rosmarinic acid, rutin, quercetin-3-O-glucoside, astragalin, apigenin-7-O-glucoside, apigenin-7-O-glucuronide and alcesefoliside, with the latter two being reported for the first time in genus Pulmonaria. The quantity of rosmarinic acid in PRE was 4.35%, distinguishing the compound as the main bioactive molecule in the species. Characterized by its high content of rosmarinic acid, P. rubra represents a highly viable candidate for subsequent development into standardized phytopharmaceuticals targeting oxidative stress-related neurodegenerative diseases. Full article

Background/Objectives: Atrasentan is a selective endothelin A receptor antagonist (SERA) developed as a potential therapy for chronic renal diseases, including diabetic nephropathy and immunoglobulin A nephropathy. Despite this potential, understanding its metabolic bioactivation is essential for assessing the risks of drug-induced liver injury (DILI). However, the metabolic profile of atrasentan remains poorly characterized, and the mechanisms underlying its potential hepatotoxicity remain underexplored. Therefore, this study aims to investigate the metabolic pathways of atrasentan in human liver microsomes (HLMs) in the presence of nicotinamide adenine dinucleotide phosphate (NADP⁺), uridine diphosphate glucuronic acid (UDPGA), or glutathione (GSH). Methods: A liquid chromatography–high resolution mass spectrometry (LC-HRMS) coupled with a feature-based molecular networking approach was used to characterize metabolites. Characterization of the major metabolites was achieved through cytochrome P450 (P450) phenotyping with human recombinant P450 isoforms. Results: A total of eighteen metabolites were characterized through phase I and II metabolic reactions, including demethylenation, N-dealkylation, O-demethylation, hydroxylation, dehydrogenation, and glucuronidation. Atrasentan acyl glucuronide (M8) was confirmed as the predominant metabolite, and we also putatively annotated a catechol intermediate (M5) and its corresponding GSH conjugate (M15). Characterizing the GSH conjugate (M15) indicates that catechol intermediate (M5) can be further oxidized to a reactive ortho-quinone intermediate, which is subsequently trapped by GSH, suggesting the potential for a bioactivation mechanism. Reaction phenotyping demonstrated that the formation of M5 is catalyzed almost exclusively by the CYP3A subfamily. However, its direct translation to in vivo oxidative stress or covalent protein binding requires further studies. Conclusions: These findings demonstrate that feature-based molecular networking is a valuable strategy for metabolite characterization, underscoring the urgent need for further in vivo metabolism studies to definitively assess hepatotoxic risks associated with these reactive metabolites. Full article

Background/Objectives: Zastaprazan (JP-1366) is a novel potassium-competitive acid blocker (P-CAB) used for the treatment of gastroesophageal reflux disease (GERD). To date, its metabolic pathways and metabolism-related drug–drug interactions (DDIs) in humans remain incompletely elucidated. This study aimed to determine the relative contributions (f_m) of cytochrome P450 isoforms to JP-1366 elimination and assess its DDI potential. Methods/Results: In vitro metabolic studies using human liver microsomes (HLMs) revealed that JP-1366 was first metabolized to M1, which subsequently underwent further oxidation, glucuronidation, and N-dealkylation. Mono-oxidation was estimated to contribute more than 46% to the overall metabolic clearance of JP-1366. Reaction phenotyping identified CYP3A as the major enzyme (f_m = 96.1%), followed by CYP1A2 (1.49%) and CYP2C9 (2.41%). By integrating in vitro data, clinical pharmacokinetic data and clarithromycin coadministration DDI data, a physiologically based pharmacokinetic (PBPK) model was developed and validated. Simulations predicted significant DDIs with strong CYP3A inhibitor (ketoconazole), with AUC ratios of 3.80. Moderate inhibitors (fluconazole and fluvoxamine) caused mild increases (AUC ratios: 1.14–1.74). Conversely, strong and moderate CYP3A inducers, rifampicin and efavirenz, produced pronounced DDIs, with AUC ratios of 0.22 and 0.50, respectively. Furthermore, simulations predicted that although JP-1366 functions as a CYP enzyme inhibitor, it would not cause clinically meaningful changes in the plasma exposure of corresponding CYP substrate drugs; however, potential interactions with CYP3A substrates still warranted consideration. Conclusions: JP-1366 is predominantly cleared via a CYP3A-dominated metabolic pathway. The PBPK simulations suggest that JP-1366 may be a moderately sensitive CYP3A substrate and a moderate inhibitor of sensitive CYP3A substrates, while its perpetrator DDI risk toward other major CYP pathways appears limited. These findings support caution or monitoring when JP-1366 is co-administered with strong CYP3A modulators or sensitive CYP3A substrates. Full article

Background/Objectives: Gold(I) complexes are promising bioactive agents with anticancer and anti-inflammatory potential. This study evaluated cytochrome P450 (CYP) interactions and in vitro pharmacokinetic properties of two Au(I)–triphenylphosphine complexes bearing 6-alkoxy-9-deazapurine ligands. Methods: Complexes [Au(HL_1,2)(PPh₃)] (HL₁ = 6-isopropyloxy-9-deazapurine, complex 1; HL₂ = 6-benzyloxy-9-deazapurine, complex 2) were investigated. Inhibition of nine human CYP isoforms was assessed in liver microsomes, and kinetics were analyzed using Dixon and Lineweaver–Burk plots. CYP binding was evaluated by UV–Vis difference spectroscopy. ADME properties (chemical/plasma stability, microsomal stability, plasma protein binding, and PAMPA permeability) were determined. Binding thermodynamics were analyzed by ITC. Results: Both complexes weakly inhibited most CYP isoforms, with stronger effects on CYP2C9 and CYP3A4/5. A non-competitive inhibition mechanism was observed, which may be related to the binding of the complexes to the substrate channels of CYP2C9 and CYP3A4, thereby limiting the active site’s accessibility to the substrate, as supported by molecular docking studies. UV–Vis spectra showed type I binding with Kd values of 9.32 µM (1) and 12.64 µM (2). Both compounds showed high chemical and plasma stability (>90%), moderate microsomal stability (~60% after 60 min), high plasma protein binding (~80%), and low passive permeability. Conclusions: Au(I)–triphenylphosphine complexes with 6-alkoxy-9-deazapurine ligands exhibit moderate CYP affinity and defined pharmacokinetic profiles, supporting further preclinical evaluation. Full article

Background/Objectives: Synthetic stimulants represent the most prevalent subclass on the new psychoactive substances (NPSs) market. However, the toxicokinetic properties of M-ALPHA, a regioisomer of MDMA and N-methyl-cyclazodone a pemoline derivative, are not yet characterized. Methods: Therefore, this study investigated the metabolism of both NPSs in pooled liver S9 fraction and rat urine, characterized cytochrome P450 (CYP) kinetics and plasma protein binding (PPB), and assessed the CYP inhibition potential of M-ALPHA, using high-performance liquid chromatography coupled to high resolution tandem mass spectrometry (HPLC-HRMS/MS). Results: Four metabolites of M-ALPHA were detected including one phase I and three phase II metabolites, resulting from demethylenation followed by subsequent methylation or glucuronidation. For N-methyl-cyclazodone, one phase I metabolite formed via N-demethylation was identified. The primary enzymes involved in M-ALPHA metabolism were CYP2B6 and CYP2D6. Notably, M-ALPHA inhibited these enzymes to a strong or moderate extent, respectively. In contrast, the metabolism of N-methyl-cyclazodone was primarily mediated by CYP2A6. PPB studies indicated low-to-moderate binding for both compounds, suggesting that significant protein-binding interactions are unlikely. Conclusions: As M-ALPHA only formed metabolites that overlapped with those of MDMA, differing only by minor retention time shifts, reliable HPLC-HRMS/MS-based identification may be challenging in clinical and forensic toxicology settings as well as doping analysis. Furthermore, drug–drug interactions following polydrug use cannot be excluded for either NPS, particularly when co-ingested with other CYP substrates metabolized by the same isoforms. Full article

Multicomponent reactions with isocyanides (IMCRs) enable the one-step assembly of complex molecules and remain a powerful strategy for accessing bioactive scaffolds. Here, we report the first synthesis of an abietane diterpene isocyanide derived from aminoimide methyl maleopimarate 1, a levopimaric acid-maleic anhydride adduct. This isocyanide was further engaged in Passerini, Ugi, and azido-Ugi reactions to provide a series of α-acyloxy- and α-acylaminocarboxamides, as well as tetrazoles, in high yields under optimized conditions. The structures of all products were confirmed by comprehensive physicochemical analysis. In silico ADME, drug-likeness, target prediction, and toxicity studies (SwissADME, ProTox-III) revealed moderate lipophilicity with favorable membrane permeability and solubility, high gastrointestinal absorption, and selective CYP3A4 inhibition with no significant effects on other CYP450 isoforms. The compounds fulfill major drug-likeness criteria, lacking undesirable reactive fragments, with only acceptable deviations in molecular weight and flexibility typical for MCR-derived products. The modifications broaden the spectrum of predicted biological targets while maintaining low overall toxicity and absence of predicted hepato- or carcinogenicity. These results demonstrate that diterpene isocyanide is a valuable building block for chemical libraries of structurally diverse abietane derivatives with peptide-like termini and highlight its potential as a source of cytotoxic, antiviral, and anti-inflammatory candidates. Full article

Background: Metabolic-dysfunction-associated steatotic liver disease (MASLD) is a growing global health burden, yet no approved pharmacological therapy currently exists. Purpose: The purpose of this study is to investigate the prophylactic and therapeutic potential of Rebamipide, a mucosal-protective and anti-inflammatory drug, in a high-fat diet (MHFD)-induced MASLD rat model, integrating quantitative liver proteomics, network analysis, and histopathology. Methods: Male Wistar rats were fed MHFD for 16 weeks and treated with Rebamipide either prophylactically (Reb T1, co-administered with diet) or therapeutically (Reb T2, administered post-NASH onset). Label-free LC-MS/MS proteomics combined with principal component analysis (PCA), partial squares discriminant analysis (PLS-DA), and enrichment analyses (including Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), Reactome via g: Profiler, network mapping, and Rat Genome Database (RGD) mining) revealed that MHFD had the following impacts: it induced the profound suppression of mitochondrial chaperones (Hspa9), microsomal triglyceride transfer protein (Mttp), and cytochrome P450 isoforms (Cyp2c6); it disrupted lipid trafficking, oxidative stress defense, and xenobiotic metabolism. Results: Rebamipide prophylaxis preserved lipid-handling proteins, prevented glycogen loss, and maintained antioxidant defenses. In contrast, therapeutic administration reversed established steatosis and remodeled metabolic pathways, enhancing fatty acid β-oxidation, detoxification, and mitochondrial protein import. Nine shared proteins across all comparisons, including MTTP and multiple Stress-70 mitochondrial isoforms, mapped to three core genes (Mttp, Cyp2c6, Hspa9) central to lipid transport, protein import, and metabolic stress adaptation. KEGG and Reactome analyses highlighted Rebamipide’s modulation of bile acid synthesis, ceramide and phosphatidylcholine metabolism, lipoprotein remodeling, and MAPK signaling. Histopathological evaluation confirmed Rebamipide’s efficacy, showing reduced steatosis and the normalization of the hepatocyte structure, with near-complete restoration in the therapeutic (Reb T2) group compared to partial protection in the Reb T1 group. Conclusions: These findings demonstrate Rebamipide’s dual-phase, multi-targeted mechanism: early protection against diet-induced metabolic injury and robust reversal of established MASLD pathology. The identified protein triad (Mttp, Cyp2c6, Hspa9) and associated pathways provide novel biomarker candidates and mechanistic insight supporting Rebamipide’s repurposing as a therapeutic for metabolic liver disease. Full article

Maclekarpine E is a minor alkaloid from Macleaya species with reported in vitro anti-inflammatory activity, but its in vivo metabolism remains unexplored. This study investigated the metabolic fate of maclekarpine E in rats and evaluated the potential pharmacological relevance of its metabolites. Maclekarpine E was orally administered to male Sprague-Dawley rats (250 mg/kg). Plasma, urine and feces were collected and analyzed by UPLC-Q-TOF-MS/MS. CYP phenotyping was performed using recombinant human enzymes. Molecular docking against ABCG2 and ABCC2 was conducted to assess potential interactions of all fecal compounds with these efflux transporters. Network pharmacology was employed to predict potential anti-ulcerative colitis-related targets of the metabolites, generating hypotheses for future experimental validation. Nineteen phase I metabolites were identified. Biotransformations included ring-opening, demethylation and oxidation. All 19 metabolites were detected in feces, nine in plasma and two in urine. No phase II conjugates were observed. CYP2C19 was the only significantly active isoform under the tested conditions, mediating approximately 16.5% substrate depletion (p < 0.05). All 20 fecal compounds bound ABCG2 (ΔG < −5.0 kcal/mol); 19 bound ABCC2. Network pharmacology yielded 57 overlapping targets with ulcerative colitis, enriched in PI3K-Akt and MAPK pathways. This study provides the first comprehensive metabolic profile of maclekarpine E in rats. The compound undergoes CYP2C19-mediated oxidation and is predominantly excreted into feces. Its fecal metabolites are potential ABCG2/ABCC2 substrates and may target UC-associated pathways based on network pharmacology predictions, warranting further experimental validation. Full article

Sex differences remain largely underexplored in metabolic disorders, particularly in the context of genetic predisposition to type 2 diabetes, the impact of aging, and environmental factors such as exposure to high-caloric diets. Previous studies using serotonin transporter (SERT)-knockout (SERT-KO) mice, which recapitulate metabolic conditions related to the lowered function of this transporter in humans, revealed an aggravated negative response of these mutants to housing on a high-fat/sugar ‘Western diet’ (WD). However, the role of sex in SERT-KO mice has not yet been studied. Available human and animal data suggest the differential regulation of insulin receptor-mediated signaling in males and females, which can be altered with aging. This study aimed to compare fat accumulation, blood biochemical changes, glucose tolerance, and insulin receptor (IR)-related signaling in the liver and various brain structures of 12-month-old male and female SERT-KO mice fed WD for 21 days. Relative to the dietary-unchallenged group and their wild-type (WT) littermates, WD-fed mutants of both sexes displayed markedly increased fat accumulation and impaired glucose and insulin tolerance. Body mass increase was more prominent in females than in males. The two sexes revealed a similar suppression of the gene expression of isoforms A and B of IR but distinct expression of IR-related factors. IR-related genes such as Cd36, Enpp, Ptpn1, Cyp4a14, Acsl1, and Pten showed differential expression between male and female SERT-KO mice fed WD. Several differences in gene expression were also found between the WT groups of the two sexes. Overall, the manifestations of hepatic steatosis, insulin resistance, and glucose tolerance were similar between the age groups of animals, whereas the gene expression of IR-related regulation differed between the groups. We conclude that aging and genetic absence of the serotonin transporter likely override sex differences in the end effects of WD challenge, while molecular mechanisms of adaptation of IR-mediated signaling are distinct between male and female SERT-KO mice fed WD. Full article

Background: Etoricoxib, a selective cyclooxygenase-2 (COX-2) inhibitor, is widely prescribed for the management of inflammatory conditions. Despite its extensive clinical use, evidence regarding its hepatic safety profile remains limited and incompletely characterized. Aims: This study aimed to systematically evaluate the hepatic effects of etoricoxib in a murine model by integrating histopathological assessment with analysis of mRNA expression of key enzymes involved in arachidonic acid metabolism Methods: Male BALB/c mice (n = 7 per group) received either low or high doses of etoricoxib (10.5 or 21 mg/kg/day) or celecoxib (35 or 70 mg/kg/day) for 28 consecutive days. Liver tissues were examined histologically using hematoxylin and eosin staining, while molecular alterations were assessed by quantitative PCR targeting representative cyclooxygenase (COX), lipoxygenase (LOX), and cytochrome P450 (CYP450) isoforms involved in arachidonic acid metabolism. Results: High-dose etoricoxib exposure was associated with pronounced hepatic histopathological alterations, including hepatocellular necrosis, inflammatory cell infiltration, and sinusoidal congestion. In contrast, low-dose treatment resulted in only mild vascular and cellular changes. At the molecular level, etoricoxib administration was associated with marked downregulation of several arachidonic acid–metabolizing genes (including Cyp4a12 and Alox12), whereas Cox2 expression was significantly upregulated (p < 0.05), indicating a shift toward a pro-inflammatory transcriptional profile. Conclusions: Etoricoxib exposure is associated with dose-dependent hepatic injury in mice, accompanied by coordinated transcriptional alterations in arachidonic acid–metabolizing pathways. Notably, molecular changes were detectable even at low doses in the absence of overt histological damage, suggesting potential early indicators of hepatic stress. These findings underscore the importance of cautious dose optimization and further translational studies to clarify the long-term hepatic safety of etoricoxib in clinical settings. Full article