芦荟大黄素通过Bax/Caspase-3信号通路调控肝癌细胞自噬作用研究  

Research of Aloe-Emodin Regulating Autophagy of Hepatoma Cells by Bax/Caspase-3 Signaling Pathway

作　　者：杜学峰[1] 朱昱[1] 徐永富[1] 杨健[1] 胡鹏[1] DU Xuefeng;ZHU Yu;XU Yongfu;YANG Jian;HU Peng

机构地区：[1]浙江省台州医院肝胆胰外科,浙江临海317000

出　　处：《新中医》2020年第20期6-9,共4页New Chinese Medicine

基　　金：浙江省基础公益研究计划项目(LQ18H160028)。

摘　　要：目的:探讨芦荟大黄素通过B淋巴细胞癌-2基因(Bcl-2)相关X蛋白(Bax)/半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)信号通路调控肝癌细胞自噬的作用。方法:选用肝癌SMMC-7721细胞,设置SMMC-7721组、5氟尿嘧啶组、芦荟大黄素低、高剂量组(低、高剂量组)。5氟尿嘧啶组加入5氟尿嘧啶至终浓度为80μg/mL,低、高剂量组加入芦荟大黄素至终浓度分别为100μg/mL、200μg/mL。以上各组每孔设6个平行样,培养72 h。试验结束后,采用MTT法测定细胞增殖水平,吖啶橙染色观察细胞自噬水平,流式细胞仪测定细胞凋亡水平,RT-PCR法及Western-blot法测定细胞Bax、Caspase-3基因和蛋白水平。结果:与SMMC-7721组比较,其余各组细胞存活率降低(P<0.05),自噬小体、凋亡率、Bax、Caspase-3 mRNA及蛋白表达水平升高(P<0.05);与低剂量组比较,高剂量组存活率降低(P<0.05),自噬小体、凋亡率、Bax、Caspase-3 mRNA及蛋白表达水平升高(P<0.05);与5氟尿嘧啶组比较,低、高剂量组细胞存活率升高(P<0.05),自噬小体、凋亡率、Bax、Caspase-3 mRNA和蛋白表达水平降低(P<0.05)。结论:芦荟大黄素能明显抑制肝癌SMMC-7721细胞增殖,诱导该细胞发生自噬和凋亡,且其机制与芦荟大黄素能促进Bax、Caspase-3 mRNA及蛋白表达有关。Objective:To discuss the effect of aloe-emodin regulating autophagy of hepatoma cells by B-cell lymphoma-2(Bcl-2)-related X protein(Bax)/Cysteine aspartic acid protease-3(Caspase-3)signaling pathway.Methods:Hepatoma SMMC-7721 cells were selected and divided into the SMMC-7721 group,the 5-fluorouracil group,as well as the aloeemodin groups of low-dose and high-dose(the low-dose group and the high-dose group).In the 5-fluorouracil group,5-fluorouracil was added to the final concentration of 80μg/mL,and in the low-dose and high-dose groups,aloe-emodin was added to the final concentration of 100μg/mL and 200μg/mL respectively.Six parallel samples were set in each group and cultured for 72 hours.After experiment,the level of cell proliferation was detected by MTT method;the level of cell autophagy was observed via acridine orange staining;the level of cell apoptosis was detected by flow cytometry;the levels of gene and protein of Bax and Caspase-3 were detected via RT-PCR and Western blot method.Results:Compared with those in the SMMC-7721 group,in other groups,the rates of cell survival were decreased(P<0.05),and the number of autophagosome,apoptosis rate,and the expression levels of mRNA and protein of Bax and Caspase-3 were increased(P<0.05).Compared with those in the low-dose group,in the high-dose group,the rate of cell survival was decreased(P<0.05),and the number of autophagosome,apoptosis rate,and the expression levels of mRNA and protein of Bax and Caspase-3 were increased(P<0.05).Compared with those in the 5-fluorouracil group,in the low-dose and high-dose groups,the rates of cell survival were increased(P<0.05),and the number of autophagosome,apoptosis rate,and the expression levels of mRNA and protein of Bax and Caspase-3 were decreased(P<0.05).Conclusion:Aloe-emodin can significantly inhibit the proliferation of hepatoma SMMC-7721 cells,induce their autophagy and apoptosis,whose mechanism is related to its promotion to the expression levels of mRNA and protein of Bax and Caspase-3.

关 键 词：肝癌 芦荟大黄素 Bax/Caspase-3信号通路 自噬 细胞实验 

分 类 号：R285.5[医药卫生—中药学]