α-常春藤皂苷协同雷公藤红素对SMMC7721肝癌细胞增殖和周期的影响  

Synergistic effects of α-hederin and celastrol on proliferation and cell cycle of SMMC7721 hepatoma cells

作　　者：张夜航 卢文丽[1] 彭佩克 方肇勤[1] ZHANG Ye-hang;LU Wen-li;PENG Pei-ke;FANG Zhao-qin(College of Basic Medical Sciences, Shanghai University of Traditional Chinese Medicine)

机构地区：[1]上海中医药大学基础医学院,上海201203

出　　处：《中成药》2022年第4期1099-1106,共8页Chinese Traditional Patent Medicine

基　　金：国家自然科学基金项目(81503482,82004000);上海市进一步加快中医药事业发展三年行动计划(2018年⁃2020年)[ZY(2018⁃2020)⁃CCCX⁃2001⁃01]。

摘　　要：目的 探索α-常春藤皂苷协同雷公藤红素对SMMC7721肝癌细胞增殖和周期的影响。方法 MTT法检测细胞存活率,CompuSyn软件获取相互作用指数(CI)并筛选出最佳协同组合。流式细胞仪检测细胞凋亡和周期分布,Western blot法检测细胞凋亡相关蛋白Bax和Bcl-2、周期蛋白依赖性激酶(CDKs)和细胞周期蛋白(Cyclins)的表达。结果 单用α-常春藤皂苷或雷公藤红素均能抑制细胞增殖,且呈剂量依赖性;作用48 h后,当α-常春藤皂苷质量浓度为10.0μg/mL,雷公藤红素质量浓度为0.30μg/mL时,CI值为0.24,协同效应最佳。在以上质量浓度条件下,协同组早期凋亡率高于其余3组(P<0.05)。α-常春藤皂苷组及协同组晚期凋亡率高于对照组(P<0.05);总凋亡率以协同组最高且高于对照组(P<0.05)。雷公藤红素及协同组G_(0)/G_(1)期比例较对照组升高(P<0.05);α-常春藤皂苷组S期比例较对照组升高(P<0.05);协同组G_(1)+S期总比例最高且高于对照组(P<0.05)。与对照组比较,各给药组Bax、Bcl-2及Bax/Bcl-2比值无明显变化(P>0.05);协同组CDK2和CDK4蛋白表达降低(P<0.05);雷公藤红素组和协同组Cyclin D1表达均升高(P<0.05);各给药组Cyclin E1表达均升高(P<0.05)。结论 α-常春藤皂苷协同雷公藤红素抑制SMMC7721细胞增殖效应确切,该效应可能与诱导G_(0)/G_(1)期周期阻滞、G_(1)+S期总比例增加有关,而Cyclin D1可能是参与其中的关键分子。AIM To explore the synergistic effects of α-hederin and celastrol on proliferation and cell cycle of SMMC7721 hepatoma cells.METHODS Results of α-hederin or celastrol treatment on the survival rate of SMMC7721 hepatoma cells determined by thiazolyl blue tetrazolium method(MTT) were processed by CompuSyn software to get combination index(CI) and to select the best synergistic combination.Further determination of apoptosis rate and cell cycle arrest by flow cytometry,and detection of the expressions of apoptosis related proteins including Bax and Bcl-2,Cyclin dependent kinase(CDKs) and Cyclins by Western blot,were carried out as well.RESULTS Being the inhibitors of cell proliferation in a concentration dependent manner,α-hederin at concentration of 10.0 μg/mL and celastrol at 0.30 μg/mL found their best synergistic effect 48 hours after treatment giving a 0.24 CI value,under which condition a higher early apoptosis rate than the other three groups was obtained(P<0.05).Higher late apoptotic rate in α-hederin and the synergistic group than that of the control group(P<0.05);and the highest total apoptotic rate in the synergistic group(P<0.05) were all observed.In contrast to the results of the control group,higher proportion of G_(0)/G_(1) phase in the celastrol group and the synergy group(P<0.05);higher proportion of S phase in the α-hederin group(P<0.05),and higher(also the highest among all the groups) total proportion of G_(1)+S phase in the synergistic group(P<0.05) were found,in addition to the nonsignificant changes in Bax,Bcl-2 and Bax/Bcl-2 ratio in each treatment group(P>0.05);decreased expressions of CDK2 and CDK4(P<0.05),increased cyclin D1 in the celastrol group and the synergistic group(P<0.05);and up-regulated cyclin E1 in all medication groups(P<0.05).CONCLUSION The good synergistic inhibitory effect on SMMC7721 cells due to the combination treatment of α-hederin and celastrol may be associated with key molecular induction of G_(0)/G_(1) phase arrest,and increase of the total proportion of G_(1

关 键 词：α-常春藤皂苷 雷公藤红素 协同作用 肝癌细胞 细胞增殖 细胞周期 Cyclin D1 

分 类 号：R285􀆰[医药卫生—中药学]