Objective: To evaluate the therapeutic potential and underlying mechanism of Latakaranja vati (LV) in dehydroepiandrosterone (DHEA)-induced polycystic ovarian syndrome (PCOS) in female Wistar rats through integrated network pharmacology, molecular docking, and experimental validation. Methods: Bioactive constituents in LV tablets were identified using liquid chromatography-mass spectrometry (LC-MS). Network pharmacology analysis was performed to predict LV-related targets and PCOS-associated genes using BindingDB, Super-PRED, GeneCards, and DisGeNET databases. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were conducted to clarify biological processes and signaling pathways. Molecular docking simulations evaluated binding affinities between LV phytoconstituents and key predicted targets. For in vivo validation, PCOS was induced in 36 female Wistar rats by daily subcutaneous administration of DHEA (60 mg/kg) for 21 d, and after successful model establishment, rats were randomly divided into DHEA, metformin (MET), clomiphene citrate (CC), and LV low-dose (LV-L, 51.5 mg/kg), medium-dose (LV-M, 103 mg/kg), and high-dose (LV-H, 206 mg/kg) groups (n = 6 each), which were orally administered for 21 d, respectively. Additional 6 rats were kept as normal control (NC) group, which did not receive any DHEA treatment. Estrous cyclicity, body weight, ovarian weight and diameter, fasting blood glucose (FBG), serum hormones [testosterone, progesterone, estrogen, follicle-stimulating hormone (FSH), luteinizing hormone (LH), and anti-Müllerian hormone (AMH)], insulin resistance [homeostatic model assessment for insulin resistance (HOMA-IR)], lipid profile [total cholesterol (TC), triglycerides (TG), high-density lipoprotein (HDL), and low-density lipoprotein (LDL)], inflammatory cytokines [tumor necrosis factor (TNF)-α and interleukin (IL)-6], oxidative stress markers [superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), malondialdehyde (MDA), and nitric oxide (NO)], myeloperoxidase (MPO), and ovarian histopathology were evaluated. Results: LC-MS analysis identified eight major phytoconstituents in LV: sitosterol, citrulline, bonducellin, oleic acid, δ-caesalpin, heptocosane, palmitic acid, and stearic acid. Network pharmacology revealed 36 overlapping targets between LV and PCOS, with key targets including estrogen receptor 1 (ESR1), nuclear receptor subfamily 3 group C member 1 (NR3C1), signal transducer and activator of transcription 3 (STAT3), and epidermal growth factor receptor (EGFR). GO and KEGG enrichment analyses indicated involvement in lipid metabolism regulation, steroid hormone receptor activity, prolactin signaling pathway, hypoxia-inducible factor (HIF)-1 signaling pathway, and insulin resistance pathways. Molecular docking demonstrated strong binding affinities between LV phytoconstituents and predicted targets, with sitosterol showing the strongest binding to EGFR (− 9.9 kcal/mol) and ESR1 (− 8.3 kcal/mol). In vivo experiments confirmed that LV treatment restored normal estrous cyclicity and significantly reduced body weight, ovarian weight, and ovarian diameter compared with DHEA group (P < 0.05, P < 0.01, or P < 0.001). LV dose-dependently restored FBG, insulin, and HOMA-IR levels (P < 0.01 or P < 0.001), and improved lipid profile, including reduced TC, TG, and LDL, and increased HDL (P < 0.05, P < 0.01, or P < 0.001). Hormonal abnormalities were corrected with testosterone, LH, and AMH decreased and progesterone, estrogen, and FSH increased (P < 0.05, P < 0.01, or P < 0.001). Furthermore, LV enhanced activities of antioxidant enzymes (SOD, CAT, and GPx), and reduced oxidative stress markers (MDA and NO) (P < 0.05, P < 0.01, or P < 0.001). Pro-inflammatory cytokines TNF-α and IL-6 were significantly suppressed, and MPO activity decreased compared with DHEA group (P < 0.05, P < 0.01, or P < 0.001). Histopathological examination showed that after LV treatment, ovarian morphology recovered with cystic follicles decreased and corpus luteum increased. Among the three LV-treated groups, LV-H group exhibited the most pronounced improvements across all parameters, indicating a clear dose-dependent therapeutic effects. Conclusion LV showed protective effects against DHEA-induced pcos by restoring endocrine balance and mitigating metabolic, oxidative, and inflammatory disturbances. The involvement of key regulatory targets, including ESR1, NR3C1, STAT3, and EGFR, supports its multi-target therapeutic potential. These findings highlight LV as a promising herbal candidate for polycystic ovarian syndrome management.

[Objective]: To investigate the therapeutic potential and underlying mechanism of Lantana camara ethanolic extract (LCEE) in ulcerative colitis (UC). [Methods] : Phytochemical analysis of LCEE was conducted using qualitative analysis, liquid chromatography-mass spectrometry (LC-MS), and high-performance thin-layer chromatography (HPTLC). The active constituents of LCEE were identified through network pharmacology analysis, followed by molecular docking. The therapeutic mechanism was validated in a UC rat model using 42 male Wistar rats (200 – 250 g) induced by 2,4,6-trinitrobenzene sulfonic acid (TNBS). Rats were randomly divided into seven groups (n = 6 per group): normal control (NC), ethanol control (EC), disease control (DC), three doses of LCEE treatment [low dose LCEE (100 mg/kg), medium dose LCEE (200 mg/kg), and high dose LCEE (400 mg/kg), p.o.], and dexamethasone (DEX, 2 mg/kg, p.o.) groups. Following TNBS-induced UC (120 mg/kg, intrarectally), rats were treated orally for 28 d. Disease severity was assessed through body weight changes, disease activity index (DAI), colon weight, colon length, and morphological scores. Haematological parameters, enzymatic antioxidants, nitric oxide (NO), myeloperoxidase (MPO), and inflammatory cytokines were measured in the serum and colon tissues. Gene expressions of tumor necrosis factor (TNF)-α, epidermal growth factor receptor (EGFR), signal transducer and activator of transcription 3 (STAT3), and B-cell lymphoma 2 (Bcl-2) were analyzed by quantitative reverse transcription polymerase chain reaction (qRT-PCR). Histopathological alterations in the colon tissues were evaluated using hematoxylin and eosin (HE), Giemsa, and periodic acid-schiff staining (PAS). [Results] : LC-MS analysis identified 13 phytoconstituents in LCEE, and HPTLC analysis confirmed the presence of ursolic acid, geniposide, and chlorogenic acid. Network pharmacological analysis identified 152 potential therapeutic targets with TNF, STAT3, Bcl-2, albumin (ALB), and EGFR as the top 5 hub targets. Molecular docking revealed strong binding affinities of LCEE phytoconstituents with key inflammatory and apoptotic targets: linaroside with TNF-α (– 6.1 kcal/mol), ursolic acid with STAT3 (– 6.8 kcal/mol) and Bcl-2 (– 8.7 kcal/mol), and cirsiliol with EGFR (– 8.2 kcal/mol), comparable to DEX. LCEE treatment significantly increased body weights and thymus weight, while significantly reducing colon weight, spleen weight, and DAI scores. Haematological parameters showed significant improvements with increased haemoglobin, red blood cells, and platelet count, and decreased white blood cells counts. Antioxidants markers were significantly improved with increased glutathione, superoxide dismutase, and catalase levels, and decreased malondialdehyde levels. LCEE significantly reduced NO and MPO levels and inflammatory cytokines including TNF-α, interleukin (IL)-1β, nuclear factor kappa-B (NF-κB), IL-6, and IL-12 compared with TNBS treated rats. LCEE downregulated the gene expression levels of TNF-α, EGFR, and STAT3, while upregulating Bcl-2 expression level, indicating modulation of inflammation and apoptosis pathways. Histological evaluation confirmed that after LECC treatment, mucosal ulcers and inflammatory cell infiltration decreased. [Conclusion] The findings suggest that Lantana camara may serve as a medicinal plant to alleviate UC and offer an investigational basis for the clinical utilization of Lantana camara.

Objective: To investigate the bone-protective potential of Nelumbo nucifera Gaertn. seed hydroalcoholic extract (NNHE) in an ovariectomized (OVX) rat model by modulating the estrogen receptor/osteoprotegerin/receptor activator of nuclear factor (NF)-κB (ER/OPG/RANKL) signaling pathway. Methods: Network pharmacology was employed with the databases of PubChem, BindingDB, DisGeNET, Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genomes (KEGG), along with Cytoscape 3.10.2 for identifying the targets and pathways of NNHE relevant to OP. A total of 48 specific pathogen-free (SPF) grade female Wistar rats were randomly divided into six groups (n = 8 per group): sham control, OVX control, OVX + NNHE [100, 200, 400 mg/(kg·d)], and OVX + alendronate [3 mg/(kg·week)]. The treatment lasted for 16 weeks. Post-treatment assessment included bone parameters (weight, thickness, density, volume, and length), serum biochemical markers [parathyroid hormone (PTH), estrogen, OPG, RANKL, tartrate-resistant acid phosphatase (TRAP), osteocalcin (OC), calcitonin (CT), calcium (Ca), phosphorus (P), and alkaline phosphatase (ALP)], pro-inflammatory cytokines [tumour necrosis factor (TNF)-α, NF-κB, interleukin (IL)-1β, and IL-6], lipid profiles [total cholesterol (TC), triglycerides (TG), low density lipoprotein (LDL), and high density lipoprotein (HDL)], oxidative stress markers [superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH), and malondialdehyde (MDA)], and histopathological analyses of femur, uterus, and vaginal tissues. Results: Network pharmacology analysis revealed 61 overlapping targets between NNHE and osteoporosis-related genes, including signal transducer and activator of transcription 3 (STAT3), NF-κB subunit 1 (NFKB1), dopamine receptor D2 (DRD2), matrix metalloproteinase 9 (MMP9), and caspase-3. GO and KEGG enrichment suggested involvement in the ER/OPG/RANKL signaling pathway. In vivo studies demonstrated that NNHE treatment (400 mg/kg) significantly reduced OVX-induced body weight gain and exhibited estrogenic activity in the vaginal cornification assay. NNHE at 200 and 400 mg/kg significantly increased serum estrogen levels compared with OVX control group, while uterine weight remained unaffected. NNHE significantly improved the lipid profile compared with OVX group, with TC, TG, and LDL decreased, while HDL levels were increased at 200 and 400 mg/kg. Bone metabolism markers were significantly improved compared with OVX group, with serum Ca and P levels restored at all NNHE doses and ALP activity reduced. NNHE effectively modulated bone turnover markers compared with OVX group by reducing levels of OC, TRAP, and PTH, and increasing level of CT. In addition, NNHE decreased RANKL level while increasing OPG level at 200 and 400 mg/kg. Bone mineral density (BMD) was significantly enhanced compared with OVX group. Serum oxidative stress was significantly mitigated compared with OVX group through increased levels of antioxidant enzymes (SOD, CAT, and GSH) and reduced MDA, with the most pronounced effects observed at 400 mg/kg. Pro-inflammatory cytokines (TNF-α, IL-6, IL-1β, and NF-κB) were significantly reduced in all NNHE treatment groups compared with OVX group. Histopathological analysis confirmed restoration of trabecular bone structure and normalization of reproductive tissue morphology in OVX rats after NNHE treatment. Conclusion NNHE demonstrated significant protective effects against OVX-induced osteoporosis through ER/OPG/RANKL signaling pathway modulation, oxidative stress, and inflammation suppression, resulting in improved BMD and structural integrity. These findings indicate that NNHE may represent a promising therapeutic candidate for postmenopausal osteoporosis management and merits further clinical investigation.

Objective: To evaluate the anti-arthritic efficacy of sarsasapogenin (SG) alone and in combination with the corticosteroid fluticasone (FC) in a rat model of rheumatoid arthritis (RA), which was induced by complete Freund’s adjuvant (CFA). Methods: Network pharmacology analysis was conducted to identify the potential molecular targets and signaling pathways of SG in RA. Targets were identified with multiple databases, including SwissTargetPrediction, GeneCards, DisGeNET, and Search Tool for the Retrieval of Interacting Genes/Proteins (STRING), and pathway enrichment analysis was performed using Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) databases. Molecular docking was performed to validate the binding affinity of key SG constituents with the predicted hub targets. Male Wistar rats were randomly divided into normal control (NC), CFA, SG, FC, and SG + FC groups (n = 6 per group). RA was induced in all groups except NC group by a single intradermal injection of CFA (0.1 mL) into the left hind paw on day 1. After successfully induction of RA (day 12), treatments were administered intradermally from day 12 to 28 as follows: SG (50 µg/rat, 40 µL per paw), FC (50 µg/rat, 40 µL per paw), or a combination of SG and FC (25 µg/rat each, total 40 µL per paw). Therapeutic outcomes were evaluated via the paw volume, joint diameter, arthritis scores, hematological and biochemical indicators, oxidative stress markers, inflammatory cytokines, and histopathological assessments of rats’ ankle joint. The gene expression analysis was performed by quantitative reverse transcription polymerase chain reaction (qRT-PCR). Acute toxicity, body weight, and immune organ indices (spleen and thymus) were also monitored to assess the potential mitigation of SG of corticosteroid-induced adverse effects. Results: Network pharmacology analysis revealed 138 potential SG-associated targets, involving 10 key hub genes. KEGG enrichment indicated the participation of pathways involving phosphoinositide-3-kinase regulatory subunit 1 (PIK3R1), estrogen receptor 1 (ESR1), E1A binding protein P300 (EP300), mammalian target of rapamycin (mTOR), C-X-C chemokine receptor type 4 (CXCR4), signal transducer and activator of transcription 3 (STAT3), and toll-like receptor 4 (TLR4). GO enrichment analysis also revealed significant involvement of inflammatory and immune-related biological processes. Molecular docking confirmed strong binding interactions between major SG constituents and the identified hub targets. SG and SG + FC groups preserved body weight, and normalized spleen and thymus indices compared with FC group (P < 0.05 or P < 0.01), suggesting the mitigation of corticosteroid-induced adverse effects. SG and SG + FC groups significantly reduced paw volume, ankle diameter, and arthritis scores compared with CFA group (P < 0.05, P < 0.01, or P < 0.001). These treatments also significantly normalized hematological indicators [red blood cells (RBC), white blood cells (WBC), hemoglobin (Hb), and platelets (PLT)] and biochemical indicators [aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (ALP)] (P < 0.05, P < 0.01, or P < 0.001). Serum proinflammatory cytokine levels [tumor necrosis factor (TNF)-α, interleukin (IL)-6, IL-12, and thromboxane B2 (TXB2)] were markedly decreased, accompanied by restored antioxidant defenses [superoxide dismutase (SOD) and glutathione (GSH)] and reduced oxidative stress markers [malondialdehyde (MDA) and myeloperoxidase (MPO)] (P < 0.05, P < 0.01, or P < 0.001). The qRT-PCR analysis demonstrated favorable downregulation of STAT3, mTOR, and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) expression levels in joint tissues in all treatment groups compared with CFA group (P < 0.05, P < 0.01, or P < 0.001). Histopathological findings corroborated these effects, indicating reduced inflammatory infiltration and preservation of joint architecture. Conclusion SG exerts protective effects against RA by modulating key inflammatory and immune pathways. The combined application of SG with FC enhances the therapeutic outcomes, while potentially reducing the corticosteroid-related adverse effects. These findings support SG as a promising adjunctive therapy in RA management, offering favorable efficacy and safety alongside conventional corticosteroid treatment.