Trophoblastic cell migration is essential for the attachment of the placenta to uterine wall, which is a critical step for fetal development. Aberrant migration of trophoblastic cells can lead to pregnant pathologies suc...Trophoblastic cell migration is essential for the attachment of the placenta to uterine wall, which is a critical step for fetal development. Aberrant migration of trophoblastic cells can lead to pregnant pathologies such as preeclampsia (PE), while the molecular mechanisms remain largely unknown. The current study investigated the effects and molecular mechanisms of the HAND1-Rac1-PAK axis in trophoblast migration using the HTR-8/SVneo cell line, isolated human primary cytotrophoblasts, and human placental tissues. We report herein that HAND1 is highly expressed in human early placenta and promotes trophoblastic cell migration. Mechanistically, HAND1 potentiates human trophoblastic cell migration via activating Vav2/Rac1/PAKs signaling, and the elevated capacity of HAND1 in inducing migration is dependent on its Serine/Threonine phosphorylation status, which is reciprocally regulated by PAK2 and is directly associated with the subsequent degradation of HAND1 protein, thereby generating a positive feed-back for HAND1-mediated trophoblasts migration during human pregnancy. The data suggest that HAND1 is essential for regulating human trophoblast function, potentially enhancing our understanding of pregnancy maintenance and contributing to new therapies for gestational disorders.
BACKGROUND: Epilepsy is a complex neurological disorder significantly influenced by genetic factors. Observational studies can identify associations, but cannot establish causality. Mendelian randomization (MR) offers a...BACKGROUND: Epilepsy is a complex neurological disorder significantly influenced by genetic factors. Observational studies can identify associations, but cannot establish causality. Mendelian randomization (MR) offers a robust tool for identifying etiology. Therefore, using multi-omics, we explored genetically proxied associations among cerebrospinal fluid metabolites, neuroimaging phenotypes, and epilepsy to prioritize candidate genes and potential therapeutic targets. METHODS: Leveraging data from genome-wide association studies, gene expression databases, and the UK Biobank, we performed two-sample MR analyses. Immune infiltration was assessed using multiple algorithms, transcription factors predicted using RcisTarget, and key genes prioritized through single-nucleotide polymorphism annotation and localization using single-cell RNA sequencing. RESULTS: We identified genetically proxied associations between 14 cerebrospinal fluid metabolites and focal epilepsy (11 protective, three risk-associated), with 13 confirmed using reverse MR. Additionally, 259 neuroimaging phenotypes showed significant associations with focal epilepsy. DAB1, ITGA8, and RORA were the top-ranked genes, localized to inhibitory neurons, oligodendrocyte precursor cells, and astrocytes, respectively. DISCUSSION: These findings demonstrate cell-specific pathology beyond standard neuronal activity. Mechanistically, DAB1 contributes to defects in inhibitory circuits, potentially disrupting excitation-inhibition balance, ITGA8 to matrix or myelin abnormalities in oligodendrocyte precursor cells, and RORA to astrocyte-mediated metabolic regulation and neuroinflammation. These robust metabolic and structural associations suggest that these features may represent early endophenotypes preceding clinical symptoms. CONCLUSION: By integrating multi-omics data, we identified a candidate gene-metabolite-imaging feature network potentially involved in epileptogenesis. The prioritized glial and neuronal targets offer hypothesis-generating insights for future mechanistic studies, precision medicine approaches, and translational research in epilepsy.
Metastasis remains the leading cause of mortality in breast cancer, and effective prevention strategies are urgently needed. Sodium ferulate (SF), a bioactive compound derived from ferulic acid, has demonstrated diverse...Metastasis remains the leading cause of mortality in breast cancer, and effective prevention strategies are urgently needed. Sodium ferulate (SF), a bioactive compound derived from ferulic acid, has demonstrated diverse pharmacological activities, yet its role in suppressing cancer metastasis remains unclear. In this study, we demonstrate that SF significantly inhibits breast cancer metastasis through comprehensive modulation of the bloodstream microenvironment. At non-toxic concentrations, SF inhibited the migration and invasion of MCF-7 and MDA-MB-231 cells by suppressing the BMP4/Smad1/5/9 signaling pathway, reversing epithelial-mesenchymal transition (EMT), and disrupting cell cycle progression. SF also enhanced the sensitivity of breast cancer cells to paclitaxel. In addition, SF reduced tumor cell adhesion to endothelial cells by blocking TNF-α-induced expression of ICAM-1 and VCAM-1 through inhibition of NF-κB signaling cascade. Furthermore, SF attenuated platelet activation and angiogenesis, two key processes involved in metastatic dissemination and colonization. In 4T1 mouse metastasis model, SF treatment significantly decreased pulmonary metastatic nodules without causing organ toxicity. Immunohistochemical analysis further confirmed decreased expression of BMP4, PI3K, and N-cadherin in lung tissues following SF treatment. Additionally, SF enhanced systemic immunity by increasing the proportion of cytotoxic T cells and natural killer cells in peripheral blood. Collectively, these findings reveal that SF exerts potent anti-metastatic effects by modulating tumor-bloodstream-immune interactions rather than direct cytotoxicity. By remodeling the bloodstream microenvironment and enhancing immune defense, SF may represent a promising and safe candidate for the prevention of breast cancer metastasis.
BACKGROUND: Necrosis of distal flap tissue is often attributed to ischemic injury. Previous research has indicated that Bergapten (BeG), known for its anti-inflammatory and antioxidant activities, protects tissues from i...BACKGROUND: Necrosis of distal flap tissue is often attributed to ischemic injury. Previous research has indicated that Bergapten (BeG), known for its anti-inflammatory and antioxidant activities, protects tissues from ischemic injury. This investigation aimed to ascertain the beneficial effects of BeG on ischemic flap survival and explore its underlying mechanisms. METHODS: To assess the survival of ischemic skin flaps, analyses of flap viability were conducted utilizing survival rate evaluations and laser Doppler blood flow (LDBF) detection. RNA sequencing was performed to clarify the underlying molecular processes involved. In addition, angiogenesis, oxidative stress (OS), pyroptosis, transcription factor EB (TFEB)-mediated autophagy, and adenosine AMP-activated protein kinase (AMPK)-transient receptor potential mucolipin 1 (TRPML1)-calcineurin (CaN) signaling were assessed using molecular docking (MD), cellular thermal shift assay (CETSA), and surface plasmon resonance (SPR), Western blot (WB) assays, immunofluorescence, and dihydroethidium (DHE) staining. RESULTS: The improvement in flap viability due to BeG was associated with the stimulation of autophagy, reduction of OS, and inhibition of pyroptosis. Notably, BeG-mediated enhancement of autophagic flux and increased resistance to OS were crucial for alleviating pyroptosis in vascular endothelial cells (VECs). BeG promoted autophagy flux and reduced endothelial oxidative stress by activating TFEB in ischemic flaps. However, the therapeutic effects of BeG were abolished by adeno-associated virus (AAV)-mediated TFEB knockdown. Additionally, BeG regulated TFEB activity through the AMPK-TRPML1-CaN pathway. CONCLUSIONS: BeG enhences autophagy and alleviates OS through stimulation of the AMPK-TRPML1-CaN-TFEB signaling cascade, hence improving the survival of ischemic flaps and potentially offering significant clinical implications.
AIM: This study investigates how the anti-Alzheimer's effectiveness of rivastigmine (RIVA) is affected by the antidiabetic drug metformin (MET). METHOD: ology: Male rats were randomly divided into a control group, an Alz...AIM: This study investigates how the anti-Alzheimer's effectiveness of rivastigmine (RIVA) is affected by the antidiabetic drug metformin (MET). METHOD: ology: Male rats were randomly divided into a control group, an Alzheimer's disease (AD) group receiving aluminum chloride (AlCl), a RIVA-treated group, a MET-treated group, and a RIVA+MET combination group. Cognitive performance was assessed using passive avoidance (PA), the radial arm maze (RAM), the Morris water maze (MWM), and novel object recognition (NOR) tests. Hippocampal microtubule-associated protein tau (MAPT), beta-site APP cleaving enzyme 1 (BACE1), acetylcholinesterase (AChE), and autophagy marker Sequestosome 1 (SQSTM1/p62) were measured, while amyloid-beta (Aβ) and caspase-3 expression were analyzed immunohistochemically. Histopathology and electron microscopy were used to assess neuronal integrity. RESULTS: MET, RIVA, and their combined treatment mitigated the neurodegenerative alterations induced by AlCl. The combination of MET+RIVA failed to yield significant differences in behavioral performance [PA, RAM, MWM, and NORT], MAPT levels, and AChE activity compared with the treatment with RIVA monotherapy. However, the combination therapy showed significant reductions in hippocampal BACE1, Aβ deposition, and SQSTM1/p62 levels, indicating enhanced suppression of amyloidogenic processing and improved autophagy. Although differences between MET+RIVA combination and RIVA monotherapy were not statistically significant for these markers, the combination markedly reduced caspase-3 immunoreactivity compared with the diseased group, indicating greater attenuation of apoptosis. CONCLUSION: These results highlight the crosstalk among the amyloid, tau, autophagy, and apoptotic pathways in AD and suggest that the MET+RIVA combination showed promising molecular improvements but no clear behavioral superiority, indicating the need for further optimization.
Allergic rhinitis (AR), a local inflammatory disease mediated by IgE in the nasal mucosa, presents a significant challenge to global health. Erucic acid (EA) is a monounsaturated fatty acid. Current research indicates EA...Allergic rhinitis (AR), a local inflammatory disease mediated by IgE in the nasal mucosa, presents a significant challenge to global health. Erucic acid (EA) is a monounsaturated fatty acid. Current research indicates EA possesses diverse biological activities, including anti-inflammatory, antioxidant, and immunomodulatory properties. However, its effects on AR remain unexplored. This study aims to investigate the therapeutic action and mechanism of EA in AR. The efficacy and mechanisms of EA were evaluated on the AR mouse model. Therapeutic outcomes were assessed through behavioral observations, measurements of various related cytokines in nasal mucosa and serum, and histopathological examination of nasal mucosa. An in vitro inflammatory model was established in RAW264.7 cells to investigate the anti-inflammatory effects and mechanisms of EA via RT-qPCR and Western blot. This study shows that, at a safe dose, EA significantly alleviates AR-associated behaviors such as rubbing and sneezing and reduces serum levels of histamine, Ovalbumin-specific IgE (OVA-sIgE), inflammatory cytokines, and chemokines. EA mitigated damage to the nasal mucosa tissue, including an increase of goblet cells, mast cells, and eosinophils. In the nasal tissue, EA also downregulated the mRNA levels of inflammation-related factors and epithelial barrier-related factors, as well as the protein levels of Myeloid Differentiation Primary Response Protein 88 (MyD88) and NF-κB. In vitro, EA suppressed Lipopolysaccharide (LPS)-induced expression of inflammatory cytokines and inhibited activation of the MyD88/NF-κB pathway. These findings demonstrate that EA exerts anti-inflammatory and epithelial barrier-restorative effects through inhibiting the MyD88/NF-κB pathway.
AIM: Intrauterine adhesions (IUA) remain a significant clinical challenge, as current surgical interventions are limited by high recurrence rates. While metabolic dysregulation is implicated in various fibrotic diseases,...AIM: Intrauterine adhesions (IUA) remain a significant clinical challenge, as current surgical interventions are limited by high recurrence rates. While metabolic dysregulation is implicated in various fibrotic diseases, its specific role in IUA pathogenesis remains largely unexplored. In this study, metabolomic analysis revealed a marked depletion of L-carnosine in IUA patients, prompting us to investigate its pathophysiological role and therapeutic potential. METHODS: We evaluated the anti-fibrotic effects of L-carnosine in vitro using TGF-β-induced human endometrial stromal cells (hESCs) and in vivo using a mouse model of IUA. Subsequently, RNA-seq was performed on hESCs to compare the TGF-β-treated group with the TGF-β plus L-carnosine group, aiming to identify differentially expressed genes and enriched signaling pathways, thereby elucidating the molecular mechanisms by which L-carnosine attenuates endometrial fibrosis. RESULTS: We demonstrated that L-carnosine treatment effectively reversed TGF-β-induced fibrosis in hESCs in vitro and attenuated uterine fibrosis in an IUA mouse model in vivo. Mechanistically, the anti-fibrotic effects of L-carnosine were mediated by the inhibition of ERK phosphorylation, which subsequently led to increased the expression of prostaglandin-endoperoxide synthase 2 (PTGS2). The functional importance of this ERK/PTGS2 axis was validated: exogenous PGE2 (the major product of PTGS2) mimicked the anti-fibrotic effects, whereas inhibition of PTGS2 with celecoxib (a specific PTGS2 inhibitor) abrogated the protective effects of L-carnosine. Conversely, pharmacological activation of ERK using mSIRK (an ERK activator) blunted the L-carnosine-induced upregulation of PTGS2. CONCLUSION: These findings identify L-carnosine depletion as a pivotal metabolic hallmark of IUA and position L-carnosine supplementation as a viable anti-fibrotic strategy by specifically targeting the ERK/PTGS2 pathway.
Fritillaria thunbergii Miq. has been traditionally used to treat respiratory disorders in Chinese medicine. Peimisine, a major F. thunbergii alkaloid, has anti-inflammatory and antioxidant effects, but its role in allerg...Fritillaria thunbergii Miq. has been traditionally used to treat respiratory disorders in Chinese medicine. Peimisine, a major F. thunbergii alkaloid, has anti-inflammatory and antioxidant effects, but its role in allergic asthma and mucus-driven airway remodeling remains unclear. This study investigated whether peimisine could attenuate asthmatic phenotypes and explored its mechanistic links to Th2 inflammation, epithelial activation, and oxidative stress. Female BALB/c mice were sensitized and challenged with ovalbumin (OVA) and treated with peimisine. Airway hyperresponsiveness (AHR) and lung function were measured, and lung tissues were evaluated for inflammatory infiltration, goblet cell hyperplasia/mucus hypersecretion, and remodeling. Inflammatory mediators and oxidative stress indices were determined in bronchoalveolar lavage fluid (BALF) and lung homogenates. In vitro, BEAS-2B bronchial epithelial cells were used to assess inflammatory activation, intracellular reactive oxygen species (ROS) generation, and monocyte-epithelial adhesion. Peimisine significantly improved AHR and reduced airway inflammatory cell infiltration. It also suppressed goblet cell expansion, mucus hypersecretion, and collagen deposition. Mechanistically, peimisine attenuated Th2-skewed inflammation, reducing BALF eosinophils, Th2 cytokines, eotaxins, and OVA-specific IgE/IgG1 while increasing Th1-associated markers and OVA-specific IgG2a. Peimisine restored redox balance and inhibited NF-κB activation in lung tissue. Consistently, peimisine downregulated epithelial eotaxins, ICAM-1, and pro-inflammatory cytokines in BEAS-2B cells, concomitant with reduced ROS production and diminished monocyte adhesion. Overall, peimisine alleviated allergic asthma by reducing airway inflammation and oxidative stress and by limiting Th2 immune activation, thereby decreasing epithelial inflammatory responses, mucus hypersecretion, and airway remodeling. These results suggest that peimisine may be a promising therapeutic candidate for allergic asthma.
UNLABELLED: Breast cancer is a leading malignancy in women. HOTAIR promotes tumor proliferation, invasion, and EMT. This study evaluated combined therapy of attenuated Salmonella typhimurium (ST) L-forms and lentiviral s...UNLABELLED: Breast cancer is a leading malignancy in women. HOTAIR promotes tumor proliferation, invasion, and EMT. This study evaluated combined therapy of attenuated Salmonella typhimurium (ST) L-forms and lentiviral shRNA-HOTAIR on triple-negative breast cancer in a murine model. METHODOLOGY: ST VNP20009 L-forms were induced by ceftriaxone. 4T1 cell invasion was assessed by Transwell Matrigel assay and apoptosis by Annexin V/PI flow cytometry in vitro and TUNEL in vivo. Tumorigenicity and aggressiveness were evaluated in a mouse breast cancer model. EGFR, Snail, and MMP9 expression in xenografts was detected by RT-qPCR and immunohistochemistry. RESULTS: ST L-forms appeared as Gram-negative bacteria with irregular red staining and long filament bodies. Transwell invasion assay showed attenuated ST and its L-forms inhibited 4T1 cell invasion in vitro. Annexin V/PI and TUNEL assays revealed that combining ST L-forms with lentiviral shHOTAIR induced significantly greater apoptosis in 4T1 cells compared to control, ST, or ST L-forms alone in vitro and in vivo. Furthermore, this combination more effectively suppressed tumor growth and aggressiveness in murine breast cancer. IHC and RT-qPCR on xenografts showed that the combination more effectively suppressed expression of EGFR, Snail, and MMP9 compared to controls. CONCLUSION: This study confirmed that the combination of attenuated S. typhimurium L-forms and lentiviral shRNA-HOTAIR significantly enhanced the suppression of tumor growth and aggressiveness in murine breast cancer. These findings supported the potential of this combined strategy of attenuated ST L-forms and lentiviral shHOTAIR as a novel and more effective bio-therapeutic approach for bacterial-mediated breast cancer therapy.
Triple-negative breast cancer (TNBC) remains one of the most aggressive breast cancer subtypes, with limited targeted treatment options. This study investigates the efficacy of folic acid (FOL) as a targeting molecule fo...Triple-negative breast cancer (TNBC) remains one of the most aggressive breast cancer subtypes, with limited targeted treatment options. This study investigates the efficacy of folic acid (FOL) as a targeting molecule for TNBC. It also examines whether the cellular uptake of FOL is influenced by acidic pH, a major feature of tumor tissue. A multifunctional FITC-Catalase-Folate (FITC-CAT-FOL) conjugate was developed and optimized to combine fluorescence tracking, enzymatic activity, and receptor targeting. Two conjugation strategies were compared to evaluate how reaction order affects labeling efficiency and catalase (CAT) stability. The optimized FITC-CAT-FOL configuration showed higher fluorescence, efficient FOL incorporation, and greater retention of catalytic activity compared to the reverse order (FOL-CAT-FITC). The conjugate maintained stable enzymatic function across different pH levels and demonstrated enhanced cellular uptake and viability at mildly acidic to neutral conditions (pH 6-7.4) through folate receptor-mediated endocytosis in TNBC cells. Fluorescence microscopy further confirmed enhanced intracellular localization of the conjugate under mildly acidic to neutral conditions. Moreover, cells treated with FITC-CAT-FOL showed higher viability and improved tolerance to oxidative stress, supporting the conjugate's biocompatibility and protective antioxidant function. These results confirm that folic acid exhibits strong affinity toward TNBC cells under physiological and tumor-relevant pH, supporting its use as an effective targeting molecule in enzyme-based drug delivery systems.
Alzheimer's disease (AD), historically characterized by amyloid plaques and tau tangles, is increasingly recognized as a disorder of intracellular signalling failure-where second messengers such as cyclic adenosine monop...Alzheimer's disease (AD), historically characterized by amyloid plaques and tau tangles, is increasingly recognized as a disorder of intracellular signalling failure-where second messengers such as cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) become silent before memory does. These cyclic nucleotides are critical for synaptic plasticity, memory consolidation, and neurovascular function, operating through tightly regulated pathways involving protein kinases such as protein kinase A (PKA) and protein kinase G (PKG), transcription factors like cAMP response element-binding protein (CREB), and phosphodiesterases (PDEs). In the healthy brain, cAMP and cGMP orchestrate gene transcription, long-term potentiation, and cerebral blood flow. However, in AD, these signalling networks are progressively disrupted. Although it is not clear, whether failures in cyclic nucleotide signalling lead to amyloid beta and tau pathology or these pathological hallmarks disrupt the signalling mechanisms and lead to synaptic failure. However, the evidence suggests both of these possibilities. Amyloid-β oligomers and tau pathology impair adenylyl and guanylyl cyclase activity, reduce CREB phosphorylation, and disrupt nitric oxide (NO) signalling. Concurrently, overexpression of PDEs accelerates cyclic nucleotide degradation, silencing downstream pathways essential for neuronal resilience and plasticity. Dysregulated cyclic nucleotide signalling has been linked to neurovascular dysfunction, and cognitive decline. The present review is significant as it reframes AD as a signalling failure disorder rather than solely a protein aggregation disease. By identifying cyclic nucleotide dysregulation as a central and druggable mechanism, it establishes a strong translational rationale for targeting PDE-mediated degradation. This mechanism-driven perspective provides a focused platform for therapeutic innovation aimed at restoring synaptic integrity and improving cognitive outcomes in AD.
β-casomorphins (BCMs) are bioactive peptides generated from milk β-casein digestion, which interact with opioid and other receptors to affect both tumor cell proliferation and immune responses within the tumor microenvir...β-casomorphins (BCMs) are bioactive peptides generated from milk β-casein digestion, which interact with opioid and other receptors to affect both tumor cell proliferation and immune responses within the tumor microenvironment. This review is aimed at summarizing current evidence on the anti-proliferative and immunomodulatory activities of BCMs, in particular BCM-5 and BCM-7, and their relevance to cancer biology. Studies have shown that BCM-5 primarily inhibits cancer cell growth, whereas BCM-7, in addition, enhances anti-tumor immunity by promoting CD8 T cell infiltration and reducing regulatory T cell populations. The review also discusses synthetic cyclic analogs designed to improve peptide stability and bioavailability, which demonstrate opioid-independent antiproliferative activities. By integration of mechanistic, in vitro, and in vivo data, this review emphasizes the potential of β-casomorphins and their derivatives as modulators of tumor progression and immune surveillance and identifies gaps in knowledge to guide future research in oncology.
Pomierny B, Krzyżanowska W, Skórkowska A
… +13 more, Jurczyk J, Przejczowska-Pomierny K, Szafarz M, Marcinkowska M, Guzda Z, Rachwał P, Kim E, Mesquita M, Torregrossa R, Whiteman M, Pomierny-Chamioło L, Pera J, Budziszewska B
Pirfenidone (PFD), a broad-spectrum anti-fibrotic agent, shows therapeutic potential in various fibrotic diseases. However, its effect on neurogenic bladder fibrosis following spinal cord injury (SCI) and its association...Pirfenidone (PFD), a broad-spectrum anti-fibrotic agent, shows therapeutic potential in various fibrotic diseases. However, its effect on neurogenic bladder fibrosis following spinal cord injury (SCI) and its association with ferroptosis remain unclear. This study aimed to investigate the therapeutic efficacy of PFD against bladder fibrosis after SCI and to preliminarily analyze its potential link with ferroptosis using a rat SCI model and a Transforming Growth Factor-β1 (TGF-β1)-induced fibrotic model in Simian Virus 40-Transformed Human Urothelial Cells (SV-HUC-1). In SCI rats, PFD treatment significantly improved urodynamic parameters, while markedly reducing collagen deposition, inflammatory infiltration, and the progression of epithelial-mesenchymal transition (EMT) in bladder tissue. In the cellular model, PFD effectively attenuated TGF-β1-induced fibrotic responses. Mechanistically, network pharmacology analysis predicted that PFD could modulate the TGF-β1 signaling and lipid peroxidation pathways. In vivo experiments confirmed that PFD reversed the dysregulation of key ferroptosis-related molecules, and in vitro studies demonstrated that PFD antagonized Erastin-induced ferroptosis, an effect comparable to the ferroptosis inhibitor Ferrostatin-1 (Fer-1). However, in the TGF-β1-induced fibrotic model, inhibition of ferroptosis alone only partially ameliorated the fibrotic phenotype, and its effect was weaker than that of PFD. This study demonstrates that PFD can effectively alleviate bladder fibrosis after SCI, and its therapeutic effect may be associated with the ferroptosis pathway. These findings suggest that PFD likely exerts its anti-fibrotic action through multi-pathway synergy, with ferroptosis inhibition representing an important contributing pathway supported by the current data, providing a new theoretical basis for treating neurogenic bladder fibrosis.
Osteoporosis (OP) is characterized by an imbalance in which bone marrow mesenchymal stem cells (BMSCs) preferentially differentiate into adipocytes rather than osteoblasts. The natural polyhydroxyflavone fisetin function...Osteoporosis (OP) is characterized by an imbalance in which bone marrow mesenchymal stem cells (BMSCs) preferentially differentiate into adipocytes rather than osteoblasts. The natural polyhydroxyflavone fisetin functions as an antioxidant and multi-target signaling regulator with potential to modulate stem cell fate. However, its role and underlying mechanisms in OP remain unclear. This study investigated whether fisetin ameliorates OP by regulating osteogenic and adipogenic differentiation using integrated in vitro and in vivo models. The effects of fisetin on BMSC differentiation were assessed using reverse transcription-quantitative polymerase chain reaction (RT-qPCR), Western blot, alkaline phosphatase (ALP)/Alizarin Red S staining, Oil Red O and lipid-droplet fluorescence staining, and immunofluorescence. RNA sequencing and network pharmacology were used to explore the underlying mechanisms, followed by validation using pharmacological and genetic approaches. The in vivo effects of fisetin were confirmed in an ovariectomy (OVX)-induced OP mouse model by micro-computed tomography (micro-CT) and bone histomorphometry. Fisetin dose-dependently promoted osteogenic differentiation and inhibited adipogenic differentiation in cultured BMSCs. Mechanistically, fisetin activated the canonical Wnt/β-catenin signaling pathway, which underpinned its pro-osteogenic and anti-adipogenic effects. In OVX-induced OP mice, fisetin treatment significantly attenuated trabecular bone loss and reduced bone marrow adiposity, demonstrating its anti-osteoporotic efficacy as evidenced by micro-CT and histomorphometric analyses. This study demonstrates that fisetin alleviates OVX-induced OP by mitigating trabecular bone loss and suppressing marrow adipogenesis via activation of the Wnt/β-catenin pathway, providing a mechanistic foundation for its development as a therapeutic agent.
Sinomenine (SIN), a bioactive alkaloid with anti-inflammatory activity, has shown therapeutic potential in ulcerative colitis (UC), but its precise molecular targets remain unclear. This study investigated the key target...Sinomenine (SIN), a bioactive alkaloid with anti-inflammatory activity, has shown therapeutic potential in ulcerative colitis (UC), but its precise molecular targets remain unclear. This study investigated the key targets and mechanisms underlying SIN-mediated protection in UC. In a dextran sulfate sodium (DSS)-induced colitis mouse model, SIN dose-dependently alleviated colitis symptoms, restored colon length, improved histopathology injury, and enhanced intestinal barrier integrity by increasing ZO-1 and claudin-1 expression. Integrated bioinformatics analyses, including differential expression analysis, WGCNA, network pharmacology, and machine-learning algorithms, identified farnesoid X receptor (FXR) as a core therapeutic target of SIN. Molecular docking predicted stable SIN-FXR binding, and cellular thermal shift assay further supported engagement of SIN with FXR. FXR expression was reduced in UC tissues and DSS-induced colitis mice but was restored after SIN treatment. Functional enrichment and single-cell RNA sequencing analyses linked FXR to lipid metabolism, immune regulation, and the Th17/Treg balance. Mechanistically, SIN suppressed arachidonic acid metabolism, reduced pro-inflammatory cytokines, increased anti-inflammatory cytokines, decreased Th17 cells, and promoted Treg accumulation. Co-administration of SIN with the FXR agonist fexaramine further potentiated its protective effects, whereas the FXR antagonist glycine-β-muricholic acid markedly reversed SIN-mediated improvements in intestinal inflammation, barrier integrity, arachidonic acid metabolism, and Th17/Treg homeostasis. Collectively, these findings demonstrate that SIN ameliorates experimental colitis, at least in part, by targeting FXR to modulate lipid metabolism and restoring Th17/Treg balance. This study provides a mechanistic basis for the therapeutic application of SIN, either alone or in combination with FXR agonists, in UC treatment.