基于谷胱甘肽/谷胱甘肽过氧化物酶4轴探讨雷公藤甲素引起肝细胞铁死亡的作用机制  

Mechanism of triptolide-induced ferroptosis in liver cells based on glutathione/glutathione peroxidase 4 axis

作　　者：王伟艳 刘珊 李会芳 常银霞 栾智华 苏越蕊 魏砚明 WANG Weiyan;LIU Shan;LI Huifang;CHANG Yinxia;LUAN Zhihua;SU Yuerui;WEI Yanming(College of Chinese Medicine and Food Engineering,Shanxi University of Chinese Medicine,Jinzhong 030619,China;Experimental Management Center,Shanxi University of Chinese Medicine,Jinzhong 030619,China)

机构地区：[1]山西中医药大学中药与食品工程学院,山西晋中030619 [2]山西中医药大学实验管理中心,山西晋中030619

出　　处：《中草药》2024年第9期2967-2975,共9页Chinese Traditional and Herbal Drugs

基　　金：山西省科技厅基础研究课题资助项目(202103021224295);山西中医药大学毒效关系研究创新团队(2022TD1016);山西省卫生健康委科研课题(2022133);山西中医药大学科技创新能力培养计划“基础研究专项”课题资助项目(2020PY-JC-17)。

摘　　要：目的观察雷公藤甲素对铁代谢和脂质过氧化的影响,探讨雷公藤甲素引起肝细胞铁死亡的可能机制。方法利用雷公藤甲素处理人正常肝细胞HL7702和C57BL/6J小鼠,或利用铁死亡抑制剂(ferrostatin-1,Fer-1)和雷公藤甲素共同处理HL7702细胞,CCK-8法检测细胞存活率;普鲁士蓝染色观察铁沉积;试剂盒检测铁离子、谷胱甘肽(glutathione,GSH)、丙二醛(malondialdehyde,MDA)含量以及超氧化物歧化酶(superoxide dismutase,SOD)、丙氨酸氨基转移酶(alanine aminotransferase,ALT)、天冬氨酸氨基转移酶(aspartate aminotransferase,AST)活性;qRT-PCR或Western blotting检测转铁蛋白受体1(transferrin receptor 1,TFR1)、谷胱甘肽过氧化物酶4(glutathione peroxidase 4,GPX4)、前列腺素内过氧化物合成酶2(prostaglandin-endoperoxide synthase 2,PTGS2)、长链脂酰辅酶A合成酶4(acyl-CoA synthetase long-chain family member 4,ACSL4)的表达。结果雷公藤甲素显著降低细胞存活率(P<0.05),引起铁沉积,升高血清中ALT、AST活性(P<0.05),增加HL7702细胞和小鼠肝组织中铁离子、MDA含量(P<0.05),降低GSH含量、SOD活性和GPX4表达(P<0.05),上调TFR1、PTGS2及ACSL4表达(P<0.05)。Fer-1显著上调细胞存活率和GPX4表达量(P<0.05),显著下调铁离子含量、TFR1、PTGS2及ACSL4表达(P<0.05)。结论雷公藤甲素可能通过GSH/GPX4轴引起铁代谢异常和脂质过氧化,诱导肝细胞铁死亡。Objective To investigate the effect of triptolide on iron metabolism and lipid peroxidation and evaluate the underlying mechanism of triptolide-induced ferroptosis in hepatic cells.Methods HL7702 cells and C57BL/6J mice were treated with triptolide,or HL7702 cells were co-treated with ferroptosis inhibitor ferrostatin-1(Fer-1)and triptolide,and then cell survival rate was detected by CCK-8 method;Iron deposition was detected by Prussian blue staining;Iron,glutathione(GSH),malondialdehyde(MDA)contents and superoxide dismutase(SOD),alanine aminotransferase(ALT),aspartate aminotransferase(AST)activities were detected by kit;Transferrin receptor 1(TFR1),glutathione peroxidase 4(GPX4),prostaglandin-endoperoxide synthase 2(PTGS2),acyl-CoA synthetase long-chain family member 4(ACSL4)expressions were detected by qRT-PCR or Western blotting.Results Triptolide significantly decreased cell viability(P<0.05),increased ALT and AST activities in serum(P<0.05),and resulted in iron deposition,increased iron,MDA contents(P<0.05),decreased GSH content,SOD activity and GPX4 expression(P<0.05),and up-regulated TFR1,PTGS2 and ACSL4 expressions in HL7702 cells and liver tissues(P<0.05).Fer-1 significantly reversed the decreased cell viability induced by triptolide,and increased GPX4 expression(P<0.05),as well as down-regulated iron content,TFR1,PTGS2 and ACSL4 expressions(P<0.05).Conclusion Triptolide may induce iron overload and lipid peroxidation via GSH/GPX4 axis which ultimately leading to ferroptosis in hepatic cells.

关 键 词：雷公藤甲素 铁死亡 肝细胞毒性 铁超载 脂质过氧化 谷胱甘肽/谷胱甘肽过氧化物酶4轴 

分 类 号：R285.5[医药卫生—中药学]