Letters in Drug Design & Discovery最新文献

{"title":"CHRM2 drives neuroblastoma progression via the PI3K/AKT pathway: mechanistic insights and Genistein-based therapeutic intervention","authors":"Zhiping Hu, Hui Liu, Liang Luo, Hengying Zhu, Ying Xu","doi":"10.1016/j.lddd.2025.100229","DOIUrl":"https://doi.org/10.1016/j.lddd.2025.100229","url":null,"abstract":"Neuroblastoma (NB) remains a challenging pediatric malignancy with limited therapeutic targets. While PI3K/AKT signaling is implicated in NB progression, novel regulators within this pathway require exploration. This study identifies CHRM2 as a critical mediator and investigates the therapeutic potential of Genistein (GST) in modulating CHRM2-driven NB pathogenesis. Integrated bioinformatics analysis identified key genes linked to PI3K/AKT signaling. CHRM2 was prioritized for functional validation using PCR, western blot, CCK-8, TUNEL, wound healing, transwell, and ELISA in SH-SY5Y cells. Gain- and loss-of-function experiments (CHRM2, si-CHRM2) and PI3K/AKT modulators (LY294002, 740 Y-P) were employed to dissect mechanistic interactions. Bioinformatics analysis revealed CHRM2 as a consistently upregulated gene in advanced-stage NB patients and a PI3K/AKT-associated hub gene. GST treatment suppressed SH-SY5Y cell viability and reduced CHRM2 expression, correlating with diminished LDH/NSE release. CHRM2 knockdown inhibited proliferation, migration, and survival while promoting apoptosis, whereas its overexpression exerted opposing effects. GST synergized with CHRM2 knockdown to amplify anti-tumor outcomes. Mechanistically, CHRM2 ablation enhanced GST-induced apoptosis via caspase 3/7 and cytochrome C activation, while PI3K/AKT pathway activation (740 Y-P) reversed GST effects. Conversely, PI3K/AKT inhibition (LY294002) exacerbated GST-mediated apoptosis and functional suppression, which was attenuated by CHRM2 overexpression. CHRM2 drives NB progression by sustaining PI3K/AKT signaling, and GST exerts anti-tumor effects via CHRM2 suppression. These findings nominate CHRM2 as a novel therapeutic target and highlight GST as a promising agent for NB treatment through PI3K/AKT pathway modulation.","PeriodicalId":18059,"journal":{"name":"Letters in Drug Design & Discovery","volume":"22 11","pages":"100229-100229"},"PeriodicalIF":0.0,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147909800","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Vitexin inhibits the malignant progression of cervical cancer by regulating CDK2 expression","authors":"Zhongyan Zhang, Zidan Luo, Bin Li, Yajun Liao","doi":"10.1016/j.lddd.2025.100218","DOIUrl":"https://doi.org/10.1016/j.lddd.2025.100218","url":null,"abstract":"Cervical cancer (CC) remains a significant global health burden, necessitating novel therapeutic strategies. Natural compounds offer promising avenues for the discovery of anticancer drug. Vitexin, a flavonoid with documented biological activities, was investigated for its potential anti-tumor effects on CC, specifically focusing on identifying its molecular targets and underlying mechanisms, which are currently poorly understood. Cell viability was analyzed by cell counting kit-8 (CCK-8) assay. Cell proliferation was analyzed by 5-Ethynyl-2’-deoxyuridine assay and flow cytometry. Cell apoptosis was analyzed by flow cytometry. Cell migration and invasion were analyzed by transwell assays. Oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) were evaluated via Seahorse XF analysis. Target prediction was performed using the GeneCards database and the Swiss Target Prediction database; common targets underwent protein-protein interaction (PPI) network analysis. Molecular analyses employed quantitative real-time polymerase chain reaction and Western blotting for mRNA and protein levels, respectively. Cycloheximide (CHX) assay was performed to assess cyclin-dependent kinase 2 (CDK2) protein stability. An immunohistochemistry assay was used to detect the positive expression rates of CDK2, nuclear proliferation marker (Ki67), hexokinase 2, and glucose transporter 1. Vitexin treatment significantly inhibited proliferation, migration, invasion, and glycolysis and induced apoptosis of HeLa and SiHa cells. Bioinformatic analysis and experimental validation identified CDK2 as a potential target protein of vitexin. In addition, CDK2 expression was significantly upregulated in clinical CC tissues compared to normal tissues and in CC cell lines versus normal cervical epithelial cells. Vitexin treatment dose- and time-dependently suppressed CDK2 protein expression and accelerated its degradation. Moreover, CDK2 knockdown inhibited CC cell proliferation, migration, invasion, and glycolysis, and promoted apoptosis. Conversely, CDK2 overexpression counteracted the anti-tumor effects of vitexin. Critically, vitexin administration suppressed the malignant growth of Siha cell xenografts in vivo by modulating CDK2 expression. Vitexin exerted potent anti-tumor effects against CC cells in vitro and in vivo primarily by downregulating CDK2. These findings position vitexin as a promising candidate for CC therapy and underscore CDK2 as a critical therapeutic target.","PeriodicalId":18059,"journal":{"name":"Letters in Drug Design & Discovery","volume":"22 11","pages":"100218-100218"},"PeriodicalIF":0.0,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147895221","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"UPLC-MS/MS and network pharmacology-based antidepressant study of Chaihu Jia Longgu Muli decoction","authors":"Huang Huang, Xiuhui Guo, Siqi Quan, Xiangli Yan, Yucheng Li, Erping Xu","doi":"10.1016/j.lddd.2025.100192","DOIUrl":"https://doi.org/10.1016/j.lddd.2025.100192","url":null,"abstract":"","PeriodicalId":18059,"journal":{"name":"Letters in Drug Design & Discovery","volume":"22 11","pages":"100192-100192"},"PeriodicalIF":0.0,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147892207","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"HS3ST2, regulated by METTL14 through m6A mechanisms, contributes to M1 macrophage polarization in chronic obstructive pulmonary disease","authors":"Dan Xu, Tao Chen, Xiaolong Ma","doi":"10.1016/j.lddd.2025.100236","DOIUrl":"https://doi.org/10.1016/j.lddd.2025.100236","url":null,"abstract":"M1-like macrophages perpetuate tissue destruction through excessive release of pro-inflammatory cytokines in chronic obstructive pulmonary disease (COPD). Heparan sulfate 3-O-sulfotransferase 2 (HS3ST2) plays a crucial role in various inflammatory disorders. However, the precise role of HS3ST2 in COPD pathogenesis remains unknown. THP-1-derived macrophages were exposed to 20 % cigarette smoke extract (CSE). Expression analysis was performed using quantitative PCR and immunoblot assays. The percentage of M1 and M2 macrophages was determined by flow cytometry. The interaction between METTL14 and HS3ST2 was confirmed by Methylated RNA immunoprecipitation (MeRIP), RIP, and luciferase assays. Bioinformatic analysis revealed elevated HS3ST2 expression in COPD pathogenesis. CSE promoted HS3ST2 upregulation and induced the M1 polarization in THP-1-derived macrophages. Knockdown of HS3ST2 attenuated CSE-induced M1 polarization in THP-1-derived macrophages. Mechanistically, METTL14 regulated HS3ST2 expression by mediating its mRNA m6A methylation. HS3ST2 restoration reversed sh-METTL14-caused suppression of M1 polarization in THP-1-derived macrophages under CSE exposure. Our study suggests that the METTL14/HS3ST2 axis functions as a novel regulator of CSE-induced M1 polarization, providing understanding of epigenetic and post-transcriptional mechanisms in COPD pathogenesis. METTL14-mediated m6A modification regulates HS3ST2 expression, thereby promoting M1 polarization in THP-1-derived macrophages. • HS3ST2 depletion attenuates CSE-induced M1 polarization in THP-1-derived macrophages. • METTL14 regulates HS3ST2 expression by mediating its mRNA m6A methylation. • The METTL14/HS3ST2 axis functions as a regulator of CSE-induced M1 polarization","PeriodicalId":18059,"journal":{"name":"Letters in Drug Design & Discovery","volume":"22 11","pages":"100236-100236"},"PeriodicalIF":0.0,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147902977","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Investigating the mechanism of luteolin in the treatment of inflammatory bowel disease based on network pharmacology, normal mode analysis and experimental validation","authors":"Yi Xin, Wenlu Chang, Qing Wang","doi":"10.1016/j.lddd.2025.100222","DOIUrl":"https://doi.org/10.1016/j.lddd.2025.100222","url":null,"abstract":"This study aimed to explore the therapeutic mechanism of luteolin in the context of inflammatory bowel disease (IBD) by integrating in vitro experiments, network pharmacology, molecular docking, and normal mode analysis. The impacts of luteolin on cell toxicity and survival were assessed by a CCK-8 assay using a LPS-induced HT-29 cell model of inflammation. Flow cytometry was utilized to analyze cell apoptosis. Levels of superoxide dismutase (SOD) and malondialdehyde (MDA) were quantified using commercially available test kits, and intracellular reactive oxygen species (ROS) were detected via flow cytometric analysis. Putative targets of luteolin were sourced from the ECTM and SuperPred databases, while genes related to IBD were acquired from the GeneCards database. Common genes between the candidate targets of luteolin and those associated with IBD were discerned by employing a Venn diagram. Subsequent functional enrichment analyses, covering both Gene Ontology (GO) terms and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, were executed using the DAVID platform. A network of protein-protein interactions (PPI) was constructed with the STRING database and graphically represented in Cytoscape software (v3.10.2) to identify central targets. Molecular docking investigations were conducted using AutoDockTools 1.5.7, and normal mode analysis was performed on the iMODS server. The results indicated that 2.0 μg/mL of luteolin inhibited cell viability (83.66 % vs 100 %, P < 0.05); therefore, subsequent experiments utilized concentrations of 0.1, 0.5, and 1.0 μg/mL. In comparison with the LPS model group, 0.5 and 1.0 μg/mL of luteolin enhanced cell viability (92.67 % vs 78.00 %, 97.00 % vs 78.00 %, P < 0.05) and SOD levels (18.00 U/mg vs 10.00 U/mg, 21.00 U/mg vs 10.00 U/mg, P < 0.05), while reducing apoptosis (15.00 % vs 29.00 %, 8.00 % vs 29.00 %, P < 0.05), MDA content (13.33 mmol/mg vs 20.00 mmol/mg, 10.50 mmol/mg vs 20.00 mmol/mg, P < 0.05), and ROS levels (27.50 % vs 41.50 %, 14.16 % vs 41.50 %, P < 0.05). No significant differences in these indices were observed with the treatment with 0.1 μg/mL of luteolin ( P > 0.05). Network pharmacology analysis identified 25 common targets for luteolin and IBD targets. These key targets were primarily associated with signaling pathways, such as the PI3K-AKT pathway, cancer, and cellular aging. Further, the results revealed a high binding affinity between luteolin and the AKT1 protein. The findings suggest that luteolin may exert its therapeutic effects on IBD by targeting AKT1 and modulating the PI3K-AKT signaling pathway, thereby inhibiting oxidative stress injury.","PeriodicalId":18059,"journal":{"name":"Letters in Drug Design & Discovery","volume":"22 11","pages":"100222-100222"},"PeriodicalIF":0.0,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147902209","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Based on network pharmacology and molecular docking to explore the mechanism that Naringin alleviates neuronal injury after cerebral ischemia/reperfusion by inhibiting ferroptosis via TP53/SLC7A11","authors":"Juanjuan Xu, Wenyu Yao, Li Li, Song Luo, Pan Lu, Jiani Hao, Dongliang Liu","doi":"10.1016/j.lddd.2025.100216","DOIUrl":"https://doi.org/10.1016/j.lddd.2025.100216","url":null,"abstract":"Acute ischemic stroke (AIS) is a global cerebrovascular disease with high disability and mortality. Previous studies have revealed the protection of Naringin against ischemia in the nervous system. This study is designed to explore the role and mechanism of Naringin in oxygen-glucose deprivation/recovery (OGD/R)-induced ischemic neuronal injury. The OGD/R model was built using SK-N-SH cells, followed by exposure to Naringin. CCK-8 and flow cytometry analyzed cell viability and apoptosis. ELISA determined TNF-α, IL-6, and IL-1β levels. Special kits detected GSH, ROS, and MDA levels. Fe + levels were determined using Iron assay kit. Western blot determined Solute carrier family 7 member 11 (SLC7A11) and tumor protein p53 (TP53) protein levels. Comparative Toxicogenomics Database (CTD), SwissTargetPrediction Database, and GeneCards Database provided targets of Naringin, AIS, and ferroptosis. PPI networks for intersected targets were constructed. KEGG enrichment analyses assessed the function of intersection targets. Molecular docking recognized the association between Naringin and core targets. Naringin relieved OGD/R-induced SK-N-SH cell apoptosis, inflammatory response, oxidative stress, and ferroptosis. From 64 targets of Naringin, AIS, and ferroptosis, PPI network identified ten hub genes. Among them, TP53 was mostly enriched in ferroptosis. Furthermore, Naringin had good binding affinity with TP53. Naringin could mitigate OGD/R-induced SK-N-SH cell damage by regulating TP53. Naringin could attenuate OGD/R-induced SK-N-SH cell injury by regulating TP53/SLC7A11, providing a new idea for AIS treatment.","PeriodicalId":18059,"journal":{"name":"Letters in Drug Design & Discovery","volume":"22 11","pages":"100216-100216"},"PeriodicalIF":0.0,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147919899","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Ginsenoside Rb1 suppresses the proliferation and tumorigenesis of U2OS osteosarcoma cells via downregulating PI3K/Akt signaling","authors":"Xing Chen, Ruicong Li, Zifeng Ruan, Qi Chen","doi":"10.1016/j.lddd.2025.100244","DOIUrl":"https://doi.org/10.1016/j.lddd.2025.100244","url":null,"abstract":"As the most prevalent malignant bone tumor, osteosarcoma (OS) primarily affects children and adolescents. Current treatment options for OS are limited by early metastasis, chemotherapy resistance, and severe side effects. Ginsenoside Rb1, a natural compound derived from Panax ginseng, has demonstrated anticancer effects on various tumor types, but its effects on OS remained to be unexplored. We examined the antitumor effect of Ginsenoside Rb1 on U2OS OS cells by cytotoxicity assay, scratch-wound healing assay, transwell assay, etc., and elucidated its underlying molecular mechanisms. Ginsenoside Rb1 significantly inhibited U2OS cell proliferation, migration, and invasion depending on the dosage while simultaneously inducing early apoptosis. Mechanistically, Ginsenoside Rb1 suppressed the PI3K/Akt signaling pathway, reducing Akt phosphorylation and downstream targets including MMP-9 and MMP-2 and cyclin D1. Ginsenoside Rb1 modulated proteins associated with apoptosis, upregulating the level of caspase-3, caspase-8, and Bax and downregulating that of Bcl-2. Ginsenoside Rb1 exerted its antitumor effects through inhibiting the PI3K/Akt signaling pathway and its downstream mediators, thereby suppressing the proliferative and metastatic behaviors of U2OS cells while promoting apoptosis. This study highlighted the potential of Ginsenoside Rb1 as a novel treatment agent against OS. However, further in vivo studies are required to assess its clinical applicability.","PeriodicalId":18059,"journal":{"name":"Letters in Drug Design & Discovery","volume":"22 11","pages":"100244-100244"},"PeriodicalIF":0.0,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147882483","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"GA inhibits CA9-mediated SMAD pathway to attenuate colon cancer metastasis and progression","authors":"Zhixin Zhang, Gang Cao, Lili Liang, Jinxuan Guo, Dandan Cheng, Zijian Zhao","doi":"10.1016/j.lddd.2026.100273","DOIUrl":"https://doi.org/10.1016/j.lddd.2026.100273","url":null,"abstract":"Colon cancer is a malignant tumor of the digestive tract associated with high morbidity and mortality. Currently, recurrence after surgery is often the ultimate cause of death in colon cancer. Gambogic acid (GA) has demonstrated anti-tumor efficacy against multiple cancers. Moreover, the expression of carbonic anhydrase 9 (CA9) expression is correlated with poor prognosis in various malignancies. Therefore, this study aims to investigate the roles and mechanisms of GA and CA9 in the progression and metastasis of colon cancer. The cellular morphology was assessed by the microscope. Cell migration and invasion were examined using wound healing and transwell assays, respectively. Angiogenesis was measured using an angiogenesis assay. Reverse-transcription quantitative polymerase chain reaction (RT-qPCR) and western blot were performed to examine the expression of protein and mRNA. Lung metastases were monitored using an in vivo live imaging system . In vivo experiments, the number of pulmonary metastatic lesions was quantified by counting metastatic nodules on the lung surface and further examined by hematoxylin and eosin (H&E) staining. GA suppressed transforming growth factor-β1 (TGF-β1)-induced colon cancer cell migration, invasion, and epithelial-mesenchymal transition (EMT), meanwhile, CA9 expression was up-regulated in colon cancer tissues and TGF-β1-stimulated colon cancer cells. Additionally, CA9 knockdown suppressed the TGF-β1-stimulated migration, invasion, and EMT by colon cancer cells. Mechanistically, GA impeded colon cancer progression and metastasis by silencing CA9 regulated small mother against decapentaplegic (SMAD) signal pathway in vivo or in vitro . GA inhibits colon cancer progression and metastasis by silencing CA9 mediated SMAD pathway. The GA may be a potential therapeutic agent for the treatment of colon cancer by regulating CA9. Gambogic acid attenuates TGF-β1-stimulated colon cancer cell migration, invasion, angiogenesis, EMT and lung metastasis by silencing the CA9 via regulating the SMAD signal pathway. • GA hampers the progression and metastasis of TGF-β1-stimulated colon cancer cells. • In the colon cancer and colon cancer cells, CA9 levels are increased. • CA9 facilitates progression and metastasis of TGF-β1-stimulated colon cancer cells. • GA inhibits the colon cancer progression and metastasis via down-regulated CA9. • GA inhibits colon cancer progression and metastasis by inhibiting CA9-regulated SMAD signal pathway.","PeriodicalId":18059,"journal":{"name":"Letters in Drug Design & Discovery","volume":"22 11","pages":"100273-100273"},"PeriodicalIF":0.0,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147883990","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Parabacteroides distasonis-derived ursodeoxycholic acid alleviates macrophage foaming and atherosclerosis progression by targeting NLRP3","authors":"Zhaofeng Wang, Yan Sun, Tao Zhu, Jiaping Li, Chengsi Qian, Mengdi Wang, Yang Fan","doi":"10.1016/j.lddd.2025.100175","DOIUrl":"https://doi.org/10.1016/j.lddd.2025.100175","url":null,"abstract":"Atherosclerosis (AS) is a persistent immune-mediated inflammatory condition characterized by high morbidity and mortality rates. Ursodeoxycholic acid (UDCA) exhibits anti-inflammatory properties and the ability to reduce lipid levels. This study aimed to investigate the action and mechanisms of Parabacteroides distasonis ( P. distasonis )-derived UDCA on AS. The anti-atherosclerotic effect of UDCA in THP-1-derived macrophages exposed to oxidized low-density lipoprotein (ox-LDLs) was investigated using Oil red O staining, biochemical analyses, qRT-PCR, western blot, and ELISA in vitro . Also, apolipoprotein E knockout (ApoE −/− ) mice fed a high-fat diet (HFD) were established to investigate the anti-atherosclerotic effects of UDCA in vivo . UDCA suppressed macrophage foaming, M1 polarization, and ox-LDL-induced NLRP3 inflammasome activation in THP-1-derived macrophages. However, NLRP3 overexpression demonstrated an opposite effect and reversed the effect of UDCA in ox-LDL-induced THP-1 macrophages. Furthermore, P . distasonis alleviated the atherosclerotic lesion area, M1 polarization, and NLRP3 inflammasome activation in HFD-fed ApoE −/− mice. UDCA alleviated ox-LDL-triggered foam cell production, M1 polarization, and inflammatory response in macrophages derived from THP-1 cells through NLRP3 inhibition.","PeriodicalId":18059,"journal":{"name":"Letters in Drug Design & Discovery","volume":"22 10","pages":"100175-100175"},"PeriodicalIF":0.0,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147879710","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"In vitro and in silico analyses reveal the potential mechanisms of Liu-Wei-Di-Huang-Wan formula for tinnitus treatment","authors":"Huan Long, Zhongjie Zeng, Tolbert Osire, Jiang Zhu, Mengfei Long","doi":"10.1016/j.lddd.2025.100179","DOIUrl":"https://doi.org/10.1016/j.lddd.2025.100179","url":null,"abstract":"Tinnitus remains a challenging auditory disorder with limited effective treatments. Liu-Wei-Di-Huang-Wan formula (LWDHW) has shown promising effects in alleviating tinnitus symptoms. This study aims to identify the key active ingredients in LWDHW and investigate their mechanisms, with a focus on the MAPK3 and STAT3 signaling pathways, to provide a scientific foundation for its application in tinnitus treatment. We utilized ultra-performance liquid chromatography–tandem mass spectrometry (UPLC-MS/MS) to analyze LWDHW constituents, identified key ingredients, and explored tinnitus treatment mechanisms through network pharmacology, Mendelian randomization, molecular docking, and molecular dynamics simulations. The cochlear cell recovery model, evaluated using HEI-OC1 cells and FACS analysis, demonstrated validated protein expressions via Western blot analysis. Additionally, computational tools were employed to assess protein-ligand interactions and pathway regulation. A total of 185 compounds were identified in LWDHW by UPLC-MS/MS, with Kaempferol, Cerevisterol, and AlisolB2 detected as active ingredients. Network pharmacology and Mendelian randomization identified AKT1, MAPK3, STAT3, and MTOR as key targets, validated by molecular docking. Fluorescence activated cell sorter (FACS) analysis revealed LWDHW’s protective effect against cisplatin-induced apoptosis, enhancing cell viability. Western blot results demonstrated that LWDHW modulates tinnitus primarily through the MAPK3 and STAT3 pathways, promoting MAPK3 phosphorylation and inhibiting STAT3 activation, thereby improving cochlear cell resilience and reducing neuroinflammation. This study identifies Kaempferol, Cerevisterol, and AlisolB2 as critical active ingredients in LWDHW, influencing key biomarkers such as AKT1, MAPK3, and STAT3. These compounds effectively regulate the MAPK3 and STAT3 pathways, offering a promising approach for enhancing tinnitus management and supporting further clinical development. • The molecular mechanisms of Liu-Wei-Di-Huang-Wan (LWDHW) in treating tinnitus were uncovered. • Active compounds and tinnitus-related targets of LWDHW were identificated and constructed. • LWDHW enhanced MAPK3 phosphorylation and inhibited STAT3 to reduce neuroinflammation.","PeriodicalId":18059,"journal":{"name":"Letters in Drug Design & Discovery","volume":"22 10","pages":"100179-100179"},"PeriodicalIF":0.0,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147897588","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}