基于生物信息学验证海桐皮-透骨草抑制炎性软骨细胞铁死亡的作用机制  

Bioinformatics-based verification of the mechanism of Haitongpi-Tougucao in inhibiting ferroptosis in inflammatory chondrocytes

作　　者：徐梦雨 吴天菊 黄璐 刘鑫 赵佳荣 游元元[1] XU Mengyu;WU Tianju;HUANG Lu;LIU Xin;ZHAO Jiarong;YOU Yuanyuan(School of Pharmacy,Chengdu Medical College,Chengdu 610500,China)

机构地区：[1]成都医学院药学院,四川成都610500

出　　处：《山东科学》2025年第1期23-31,共9页Shandong Science

基　　金：国家中医药管理局全国名老中医药专家传承工作室建设项目(国中医药人教函[2019]41号);成都医学院应用开发与成果转化培育项目(CYCG19-01);四川省大学生创新训练项目(S202313705037)。

摘　　要：基于生物信息学验证海桐皮-透骨草(复方海桐皮)抑制脂多糖(LPS)诱导的大鼠炎性软骨细胞铁死亡的作用机制。通过生物信息学工具预测骨关节炎铁死亡的相关作用机制,确定待验证通路。利用相关试剂盒检测亚铁离子浓度、还原型谷胱甘肽(GSH)质量分数;利用酶联免疫法(ELISA)检测加药后细胞活力和相关细胞因子IL-1β、IL-6、TNF-α的质量浓度;蛋白免疫印迹法(Western-Blot)检测各组NLRP3炎症小体通路相关的蛋白NLRP3、Caspase-1、ASC以及抑制铁死亡的基因GPX4蛋白表达水平。结果发现亚铁离子浓度显著降低,GSH质量分数显著升高;ELISA实验结果显示,药物各组与模型组相比IL-1β、IL-6、TNF-α炎症因子质量浓度均降低;WB结果显示,药物各剂量组与模型组相比,NLRP3、Caspase-1、ASC蛋白表达水平显著降低,GPX4蛋白表达水平显著升高。复方海桐皮可以通过介导NLRP3炎症小体通路对炎性软骨细胞的铁死亡进行干预,从而达到治疗骨关节炎的目的。Based on bioinformatics,this study validates the mechanism of action of Haitongpi-Tougucao(compound Haitongpi)in inhibiting lipopolysaccharide(LPS)-induced ferroptosis in rat inflammatory chondrocytes.Bioinformatics tools were used to predict the mechanism of action of ferroptosis in osteoarthritis and identify pathways for validation.The key techniques used were as follows:the detection of ferrous ion content and reduced glutathione(GSH)content using relevant kits;the detection of cell viability and the levels of related cytokines IL-1β,IL-6,and TNF-αusing the enzyme-linked immunosorbent assay(ELISA)after dosing;and the use of protein immunoblotting(western blot,WB)to detect the protein expression levels of NLRP3,Caspase-1,ASC,and GPX4,a gene that inhibits ferroptosis,related to the NLRP3 inflammasome pathway in each group.The results revealed that the ferrous ion content was significantly decreased,while the GSH content was significantly increased;the ELISA experiment showed that the levels of inflammatory factors IL-1β,IL-6,and TNF-αwere decreased in each group administered with the drug compared with those in the model group;the WB results showed that the expression levels of NLRP3,Caspase-1,and ASC proteins were significantly decreased and GPX4 protein expression levels were significantly increased in each group administered with a specific dosage of the drug compared with those in the model group.Therefore,the compound Haitongpi can intervene in the ferroptosis of inflammatory chondrocytes by mediating the NLRP3 inflammasome pathway,thereby achieving the purpose of osteoarthritis treatment.

关 键 词：骨关节炎 铁死亡 海桐皮-透骨草 生物信息学 NLRP3炎症小体通路 

分 类 号：R285[医药卫生—中药学] R289[医药卫生—中医学]