基于网络毒理学和动物实验研究苦参碱的肝毒性机制OA
Study on the hepatotoxicity mechanism of matrine based on network toxicology and animal experiments
目的 基于网络毒理学、动物实验探讨苦参碱肝毒性的作用机制.方法 通过TCMS和CTD等数据库搜集苦参碱肝毒性靶点,利用GeneCards数据库得到肝毒性疾病相关靶点.取两者交集靶点,上传至STRING数据库构建靶点互作图,DAVID(https://david.ncifcrf.gov/home.jsp)数据库进行GO功能注释分析和KEGG通路富集分析,并进行分子对接.小鼠随机分为阴性对照组、苦参碱组(200 mg/kg)、对乙酰氨基酚组(300 mg/kg),每组10只,单次灌胃给药.通过HE染色、DAPI染色,检测血清丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、碱性磷酸酶(ALP)、乳酸脱氢酶(LDH)、超氧化物歧化酶(SOD)、还原型谷胱甘肽(GSH)水平进行体内实验验证,qPCR法检测肝脏组织TNF、IL-6等基因的mRNA表达量.结果 网络毒理学共筛选出130个苦参碱和肝脏毒性交集靶点.KEGG共富集出165条通路,涉及炎症介质释放、TNF信号通路、TGF-β1信号通路等;分子对接结果表明,苦参碱对TNF等核心基因具有较好的亲和性;动物实验结果表明,苦参碱组小鼠SOD、GSH含量显著下降(P<0.05或P<0.01)),肝脏组织出现明显损伤,qPCR研究结果进一步表明,苦参碱能显著上调TNF、IL6、TGF-β1、TP53和NOS2基因表达,下调NQO1基因表达水平.结果 提示苦参碱可诱发氧化应激、诱导细胞凋亡.结论 苦参碱诱导的肝毒性是多靶点、多通路协同调控的结果,其分子机制可能与TNF、TGF-β1 信号通路的异常调控相关,进而介导肝细胞损伤、炎症反应及肝组织病理改变.
Objective To explore the mechanism of matrine hepatotoxicity based on network toxicology and animal experiments.Methods The TCMS and CTD databases were used to collect the targets of matrine hepatotoxicity,and the GeneCards database was used to obtain the targets related to hepatotoxic diseases.The intersection targets of the two sets were taken and uploaded to the STRING database to construct the target interaction map.The DAVID(https://david.ncifcrf.gov/home.jsp)database was used for GO functional annotation analysis and KEGG pathway enrichment analysis,and molecular docking was performed.The mice were randomly divided into a negative control group,a matrine group(200 mg/kg),and an acetaminophen group(300 mg/kg),with 10 mice in each group,and received a single gavage administration.In vivo experiments were conducted to verify the levels of serum alanine aminotransferase(ALT),aspartate aminotransferase(AST),alkaline phosphatase(ALP),lactate dehydrogenase(LDH),superoxide dismutase(SOD),and glutathione(GSH)through HE staining and DAPI staining.The mRNA expression levels of genes such as TNF and IL-6 in liver tissues were detected using qPCR.Results A total of 130 matrine and hepatotoxicity intersection targets were screened out by network toxicology.A total of 165 pathways were enriched by KEGG,involving the release of inflammatory mediators,TNF signaling pathway,TGF-β1 signaling pathway,etc.Molecular docking showed that matrine had good affinity for core genes such as TNF.The results of animal experiments showed that the contents of SOD and GSH in the matrine group were significantly decreased(P<0.05 or P<0.01),and the liver tissue was significantly damaged.The results of qPCR further showed that matrine significantly upregulates the expression of TNF,IL-6,TGF-β1,TP53 and NOS2 genes,and downregulates the expression of NQO1 gene.The results showed that matrine may induce oxidative stress and induce apoptosis.Conclusion Matrine-induced hepatotoxicity is the result of multi-target and multi-pathway synergistic regulation,and its molecular mechanism may be related to the abnormal regulation of TNF and TGF-β1 signaling pathways,which in turn mediates hepatocyte damage,inflammatory response,and pathological changes in liver tissue.
张红妹;陈杰君;杜青云;杨仁轩;吴俊标;郭秋平
广州中医药大学第二临床医学院,广东 广州 510006广州医药研究总院有限公司,广东 广州 510245广州医药研究总院有限公司,广东 广州 510245深圳平乐骨伤科医院,广东 深圳 518010广州中医药大学深圳医院,广东 深圳 518034广州医药研究总院有限公司,广东 广州 510245
医药卫生
苦参碱肝毒性网络毒理学分子对接TNF信号通路TGF-β1信号通路
matrinehepatotoxicitynetwork toxicologymolecular dockingTNF signaling pathwayTGF-β1 signaling pathway
《广东药科大学学报》 2026 (2)
64-76,13
广东省自然科学基金项目(2024A1515012010)
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