红景天苷对HepG2细胞增殖、迁移、侵袭及凋亡的影响  

Effect of Salidroside on Proliferation,Migration,Invasion,and Apoptosis of HepG2 Cells

作　　者：蒋兵 杨韬[1] 封龙飞 王涛[2] 苏海翔[1,2] JIANG Bing;YANG Tao;FENG Longfei;WANG Tao;SU Haixiang(Gansu University of Chinese Medicine,Lanzhou 730000,China;Gansu Provincial Academic Institute for Medical Research,Gansu Provincial Cancer Hospital,Lanzhou 730050,China)

机构地区：[1]甘肃中医药大学,兰州730000 [2]甘肃省医学科学研究院,甘肃省肿瘤医院,兰州730050

出　　处：《中国实验方剂学杂志》2022年第17期75-83,共9页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：甘肃省科技计划项目(18JR2FA010);甘肃省省级重点人才项目(甘组通字[2021]17号);甘肃省中医药科研课题(GZKP-2021-21)。

摘　　要：目的:红景天苷是从红景天的根茎部中提取得到的,是红景天中含量最高的天然活性化合物。该实验旨在研究红景天苷对人源性肝癌细胞(HepG2细胞)增殖、迁移、侵袭和凋亡的影响。方法:选取未作任何处理的HepG2细胞作为空白组,以终浓度为20、40、80μmol·L-1红景天苷处理的HepG2细胞作为红景天苷组。采用多功能细胞分析仪测定红景天苷对HepG2细胞增殖的影响,划痕实验测定红景天苷对HepG2细胞迁移能力的影响,Transwell小室实验测定红景天苷对HepG2细胞侵袭能力的影响,倒置显微镜观察红景天苷对HepG2细胞形态的影响,透射电镜观察红景天苷对HepG2细胞中线粒体的影响,流式细胞术分析红景天苷对HepG2细胞凋亡及周期分布的影响,实时荧光定量聚合酶链式反应(Real-time PCR)测定红景天苷对HepG2细胞相关凋亡基因的影响,蛋白免疫印迹法测定红景天苷对HepG2细胞相关迁移、侵袭及凋亡蛋白的影响。结果:与空白组比较,红景天苷组(20、40、80μmol·L-1)细胞指数均明显下降(P<0.05),愈合面积均明显增加(P<0.05),且呈时间和浓度依赖性;红景天苷组(20、80μmol·L-1)中能够穿过Matrigel基质胶的HepG2细胞数量呈浓度依赖性减少;红景天苷组(20、40、80μmol·L-1)总凋亡率呈浓度依赖性升高,且阻滞细胞于G2/M期(P<0.05);红景天苷组(20、80μmol·L-1)上皮性钙黏附蛋白(E-cadherin)表达呈浓度依赖性升高(P<0.05),神经钙黏附蛋白(N-cadherin)表达呈浓度依赖性降低(P<0.05)。红景天苷组(20、40、80μmol·L-1)胱天蛋白酶-3(Caspase-3)mRNA表达、B细胞淋巴瘤-2(Bcl-2)相关X蛋白(Bax)、Caspase-3蛋白及胱天蛋白酶-9(Caspase-9)蛋白表达均呈浓度依赖性升高(P<0.05),肌动蛋白结合蛋白(Girdin)及Bcl-2表达均呈浓度依赖性降低(P<0.05)。结论:红景天苷对HepG2细胞的增殖、迁移及侵袭均具有抑制作用,且通过线粒体途径诱导HepG2细胞凋亡。表明红�Objective:Salidroside is the most abundant natural active compound in the famous Chinese herbal medicine Rhodiolae Crenulatae Radix et Rhizoma.This study aims to explore the effect of salidroside on the proliferation,migration,invasion,and apoptosis of human hepatoma(HepG2)cells.Method:The HepG2cells without any treatment were selected as the blank group,and the HepG2 cells in the salidroside groups were treated with salidroside at final concentrations of 20,40,80 μmol·L-1,respectively.A multifunctional cell analyzer,scratch assay,and Transwell assay were employed to determine the proliferation,migration,and invasion of HepG2 cells,respectively.An inverted microscope was used to observe the morphology,and a transmission electron microscope to observe the mitochondria of HepG2 cells.Flow cytometry was employed to determine the apoptosis and cycle distribution of HepG2 cells.Real-time fluorescent quantitative polymerase chain reaction( Real-time PCR) and Western blot were employed to determine the expression of apoptosisassociated genes and migration-,invasion-,and apoptosis-associated proteins,respectively,in HepG2 cells.Result:Compared with the blank group,salidroside(20,40,80 μmol·L-1)decreased the cell index and increased the healing area in a time-and dose-dependent manner(P<0.05).Compared with that in the blank group,the HepG2 cells that could pass through Matrigel reduced in the salidroside(20,80 μmol·L-1)groups.Compared with the blank group,salidroside(20,40,80 μmol·L-1)increased the total apoptosis rate in a dose dependence manner and blocked the cells in the G2/M phase(P<0.05).Compared with the blank group,salidroside up-regulated the expression of epithelial-cadherin(E-cadherin)in a dose-dependent manner(P<0.05)and down-regulated that of nerve-cadherin(N-cadherin)in the 20 and 80 μmol·L-1groups(P<0.05).Compared with the blank group,salidroside(20,40,80 μmol·L-1)up-regulated the mRNA level of cysteinecontaining aspartate-specific protease-3(Caspase-3)and the protein levels of B-cell lymphoma-2

关 键 词：肝癌 红景天苷 增殖 迁移 侵袭 凋亡 

分 类 号：R22[医药卫生—中医基础理论] R242[医药卫生—中医学] R2-031R285.5