龙葵提取物澳洲茄碱诱导A549细胞凋亡的机制研究  

Molecular Mechanism of Apoptosis Induced by Solasonine Extracted from Solanum nigrum in A549 Cells

作　　者：李龙妹 黄锦鹏 河文峰 李秋萍 吴万垠 龙顺钦 LI Longmei;HUANG Jinpeng;HE Wenfeng;LI Qiuping;WU Wanyin;LONG Shunqin(The Second Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510370 Guangdong,China)

机构地区：[1]广州中医药大学第二附属医院,广东广州510370

出　　处：《中药新药与临床药理》2020年第12期1422-1427,共6页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金青年项目(81803919);广东省自然科学基金博士启动项目(2018A030310601)。

摘　　要：目的探讨龙葵提取物澳洲茄碱(Solasonine,SS)对人肺癌A549细胞增殖、凋亡的影响及相关的分子机制。方法将浓度为0、15、20、25μmol·L^-1的澳洲茄碱分别作用于A549细胞24、48、72 h,将15μmol·L^-1的顺铂(DDP)及25μmol·L^-1的澳洲茄碱分别作用于A549、PC9、H1650细胞24 h,采用MTT法检测细胞存活率。将15、20、25μmol·L^-1的澳洲茄碱及15μmol·L^-1的顺铂作用于A549细胞24 h,采用Annexin V-FITC/PI流式细胞术检测细胞凋亡率,采用Western Blot法检测半胱天冬酶3酶原(proCaspase-3)、聚腺苷二磷酸-核糖聚合酶(PARP)、B淋巴细胞瘤-2(Bcl-2)、Bcl-2相互作用杀伤蛋白(Bik)、拟南芥类受体激酶(Bak)和核因子κB p65蛋白的表达。结果与空白对照组比较,澳洲茄碱15、20、25μmol·L^-1给药组在作用A549细胞24、48、72 h的细胞存活率均明显降低(P<0.05);作用24 h后,澳洲茄碱25μmol·L^-1组及顺铂15μmol·L^-1组的A549、PC9、H1650细胞存活率均明显降低(P<0.05);澳洲茄碱20、25μmol·L^-1组及顺铂15μmol·L^-1组的正常细胞占比降低,细胞早期凋亡占比和晚期凋亡占比增加,细胞凋亡率明显升高(P<0.05);澳洲茄碱15、20、25μmol·L^-1组A549细胞的proCaspase-3蛋白表达明显下调(P<0.05);澳洲茄碱20、25μmol·L^-1组A549细胞的Bcl-2、p65、PARP蛋白表达明显下调,而Bik、Bak蛋白表达明显上调,差异均有统计学意义(P<0.05)。结论澳洲茄碱可能通过抑制p65、Bcl-2蛋白表达,并增强Bik、Bak蛋白表达,激活Caspase-3通路,进而诱导A549细胞凋亡。Objective To explore the effects of Solasonine on the proliferation and apoptosis in human lung cancer A549 cells and the related molecular mechanism.Methods The Solasonine with concentrations of 0,15,20,25μmol·L^-1 were added to the A549 cells for 24,48,and 72 hours,respectively.The DDP with a concentration of 15μmol·L^-1 and the Solasonine with a concentration of 25μmol·L^-1 were added to the A549,PC9,and H1650 cells for 24 hours,respectively.Then the cell viability was detected by MTT assay.The Solasonine with concentrations of 15,20,25μmol·L^-1 and the DDP with a concentration of 15μmol·L^-1 were added to the A549 cells for 24 hours,respectively.Then the apoptosis rate was detected by Annexin V-FITC/PI flow cytometry analysis.The expression of proCasapse-3,PARP,Bcl-2,Bik,Bak and p65 were detected by Western Blot assay.Results Compared with the blank control group,the cell viabilities in Solasonine groups with concentrations of 15,20,25μmol·L^-1 were reduced significantly after treating A549 cells for 24,48,and 72 hours(P<0.05).The cell viabilities in Solasonine group with a concentration of 25μmol·L^-1 and the DDP group with a concentration of 15μmol·L^-1 were reduced significantly after treating A549,PC9 and H1650 cells for 24 hours(P<0.05).The proportion of normal cells was decreased,and the proportions of early apoptosis and late apoptosis were increased,and the apoptosis rates were significantly increased in the solasonine groups with concentrations of 20,25μmol·L^-1 and the DDP group with a concentration of 15μmol·L^-1(P<0.05).The expression of proCaspase-3 in solasonine groups with concentrations of 15,20,25μmol·L^-1 were reduced significantly(P<0.05).The expression of PARP,Bcl-2 and p65 in solasonine groups with concentrations of 20,25μmol·L^-1 were reduced significantly(P<0.05),but the expression of Bik and Bak were increased significantly(P<0.05).Conclusion The apoptosis induced by Solasonine is achieved by inhibiting the expression of p65 and Bcl-2,enhancing the expression of

关 键 词：龙葵 澳洲茄碱 A549细胞 凋亡 半胱天冬酶3 B淋巴细胞瘤-2家族 核因子κB p65 

分 类 号：R285.5[医药卫生—中药学]