雷公藤甲素联合阿霉素对人肝癌Huh7细胞凋亡的影响  

Effects of Triptolide Combined with Doxorubicin on the Apoptosis of Hepatocellular Carcinoma Huh7 Cells

作　　者：彭宵 李强[1] 田丽莉 朱国福[1] PENG Xiao;LI Qiang;TIAN Lili;ZHU Guofu(School of Pharmacy,Shanghai University of Traditional Chinese Medicine,Shanghia2i01203;Traditional Chinese Medicine Pharmacy,Zhejiang Hospital,Hangzhou 310013)

机构地区：[1]上海中医药大学中药学院,上海201203 [2]浙江医院中药房,杭州310013

出　　处：《中药药理与临床》2023年第8期52-57,共6页Pharmacology and Clinics of Chinese Materia Medica

基　　金：上海中医药大学钱伯文名师工作室项目(编号:A1-C21-205-0106);浙江省中医药优秀青年人才项目(编号:2020ZQ001)。

摘　　要：目的:探究雷公藤甲素联合阿霉素对人肝癌Huh7细胞增殖、凋亡的影响。方法:将Huh7细胞分为正常对照组、雷公藤甲素30 nmol/L组、阿霉素2μmol/L组、联合给药(雷公藤甲素30 nmol/L+阿霉素2μmol/L)组,采用MTT法检测各组Huh7细胞存活率;Hoechst 33258染色和流式细胞术检测细胞凋亡;Western blot法检测凋亡相关蛋白及DNA损伤修复相关蛋白的表达。结果:与正常对照组相比,2μmol/L阿霉素可抑制Huh7细胞增殖,30 nmol/L雷公藤甲素对Huh7细胞的增殖无明显抑制作用(P>0.05);与阿霉素2μmol/L组相比,雷公藤甲素30 nmol/L+阿霉素2μmol/L组可显著抑制Huh7细胞的增殖(P<0.01),且联合给药组对Huh7细胞的凋亡诱导作用明显增强(P<0.01),Huh7细胞中促凋亡蛋白Cleaved Caspase 3、BAX及DNA损伤标志蛋白γ-H2AX表达显著上调(P<0.01),抗凋亡蛋白BCL-2及DNA损伤修复蛋白XRCC1、PARP、Rad51的表达显著下调(P<0.05或P<0.01)。结论:30 nmol/L雷公藤甲素可显著增强阿霉素对Huh7细胞的增殖抑制作用、凋亡诱导作用,其机制可能与上调Cleaved Caspase 3、BAX表达,下调BCL-2表达,抑制DNA损伤修复反应有关。Objective:To explore the effects oftriptolide combined with doxorubicin on the proliferation and apoptosis of human hepatocellular carcinoma Huh7 cells.Methods:Huh7 clls were divided into a normal control group,a triptolide group(30 nmol/L),a doxorubicin group(2μmol/L),and a combination group(30 nmol/L triptolide+2μmol/L doxorubicin).The cell viability of Huh7 cells in each group was detected by MTT assay.Cell apoptosis was detected by Hoechst 33258 staining and flow cytometry.Western blot was used to detect the expression of proteins related to apoptosis and DNA damage repair.Results:Compared with the conditions in the normal control group,2μmol/L doxorubicin inhibited the proliferation of Huh7 cells,while 30 nmol/L triptolide had no significant effect on the proliferation of Huh7 cells(P>0.05).Compared with 2μmol/L doxorubicin,triptolide(30 nmol/L)+doxorubicin(2μmol/L)significantly inhibited the proliferation of Huh7 cells(P<0.01)and significantly enhanced the induction of apoptosis of Huh7 cells(P<0.01).In addition,protein expression of pro-apoptotic proteins Cleaved Caspase-3,Bax,and DNA damage marker protein-H2AX was up-regulated in Huh7 cells in the combination group(P<0.01),while the expression of anti-apoptotic protein Bcl-2 and DNA damage marker proteins XRCC1,PARP,and Rad51 was down-regulated(P<0.05 or P<0.01).Conclusion:The 30 nmol/L triptolide can significantly enhance the inhibitory effect of doxorubicin on proliferation and apoptosis induction of Huh7 cells,and the mechanism may be attributed to up-regulating Cleaved Caspase 3 and Bax expression,down-regulating Bcl-2 expression,and inhibiting DNA damage repair response.

关 键 词：雷公藤甲素 阿霉素 HUH7细胞 凋亡 DNA损伤 

分 类 号：R735.7[医药卫生—肿瘤]