吴茱萸碱诱导人胃腺癌细胞MGC-803程序性坏死的作用机制  

Mechanism of evodiamine inducing necroptosis in human gastric adenocarcinoma cell line MGC-803

作　　者：刘丽平 李德亮 李峰 李琴[1,4] 葛科立 LIU Li-ping;LI De-liang;LI Feng;LI Qin;GE Ke-li(Institute of Integrated Medicine,School of Basic Medicine,Qingdao University,Qingdao 266023,China;Affiliated Hospital of Qingdao University,Qingdao 266000,China;Zibo Bashan Wanjie Hospital,Zibo 255200,China;Shenzhen University General Hospital,Shenzhen 518000,China)

机构地区：[1]青岛大学基础医学院中西医结合中心,青岛266023 [2]青岛大学附属医院,青岛266000 [3]淄博岜山万杰医院,淄博255200 [4]深圳大学总医院,深圳518000

出　　处：《中华中医药杂志》2022年第4期2259-2263,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：山东省重点研发计划(No.2017GSF218084);青岛市应用基础专项(No.19-6-2-31-cg)。

摘　　要：目的:探究吴茱萸碱(EVO)诱导人胃腺癌细胞MGC-803程序性坏死的作用机制。方法:CCK-8法检测细胞活力;DCFH-DA法、Mito-Sox法、Flou-4 AM法、JC-1法分别检测活性氧簇(ROS)水平、Ca^(2+)水平和线粒体膜电位;Western Blot检测蛋白表达。结果:EVO能明显诱导MGC-803细胞活力下降,呈时间-剂量依赖性(P<0.01);EVO(2.5μmol/L)处理细胞6、12、24 h后,ROS和Ca^(2+)水平明显增加、线粒体膜电位明显下降,受体相互作用蛋白激酶(RIP)1、RIP3和混交激酶域蛋白(MLKL)磷酸化水平上升;EUK-134、鱼藤酮(Rotenone)、BAPTA-AM和辣椒平预处理可减轻EVO诱导的细胞活力、线粒体膜电位下降,ROS水平、Ca^(2+)水平和RIP1、RIP3和MLKL磷酸化水平上升。结论:EVO可通过辣椒素受体(TRPV)1/Ca^(2+)/ROS激活RIP1/RIP3/MLKL信号通路诱导MGC-803细胞程序性坏死。Objective:To investigate the mechanism of evodiamine(EVO)inducing necroptosis in human gastric adenocarcinoma cell line MGC-803.Methods:CCK-8 assay was used to detect cell viability.Reactive oxygen species(ROS)level,Ca^(2+)level and mitochondrial membrane potential were detected by DCFH-DA assay,Mito-Sox assay,Flou-4 AM assay and JC-1 assay.Protein expression was detected by Western Blot.Results:EVO could significantly inhibit MGC-803 cell viability in a time-dose dependent manner(P<0.01).After EVO(2.5μmol/L)treatment for 6,12,24 h,ROS and Ca^(2+)levels significantly increased,mitochondrial membrane potential significantly decreased,and phosphorylation of RIP1,RIP3 and MLKL were increased.EUK-134,Rotenone,BAPTA-AM and capsazepine could alleviate the effects of EVO causing the decresion of cell viability,mitochondrial membrane potential and the increasing of ROS levels,Ca^(2+)levels,RIP1,RIP3,and MLKL phosphorylation.Conclusions:EVO induced necroptosis in MGC-803 cells by activating RIP1/RIP3/MLKL signaling pathway through TRPV1/Ca2+/ROS.

关 键 词：吴茱萸碱 胃癌 程序性坏死 活性氧簇 Ca^(2+) 辣椒素受体 

分 类 号：R285[医药卫生—中药学]