摘要
目的：研究党参总皂苷（TSC）对人肝癌SMMC-7721细胞的抑制作用,并初步探讨其作用机制。方法：采用MTT法检测0、100、200、400、600、800、1 000、1 200、1 400、1 600、1 800、2 000 mg/L的TSC作用于SMMC-7721细胞72 h后的细胞活力,并计算生长抑制率和半数抑制浓度（IC50）;采用流式细胞仪检测911.0 mg/L的TSC作用于SMMC-7721细胞24、48、72 h后的细胞凋亡情况并计算凋亡率;采用酶标仪检测0（空白对照）、200、400、800、1 000 mg/L的TSC作用于SMMC-7721细胞72 h后半胱氨酸蛋白酶3（Caspase-3）、Caspase-8、Caspase-9活性,并采用双抗体夹心酶联免疫法检测其p38丝裂原活化蛋白激酶（MAPK）、p53蛋白表达。结果：100-2 000 mg/L TSC可抑制SMMC-7721细胞的增殖,且与TSC给药浓度呈正相关,其IC50为911.0 mg/L;911.0 mg/L的TSC作用于细胞24、48、72 h的凋亡率分别为21.10%、30.20%、41.10%。与空白对照比较,200、400、800、1 000 mg/L的TSC能增强细胞中Caspase-3、Caspase-8、Caspase-9活性与p38MAPK、p53蛋白表达,差异具有统计学意义（P〈0.01或P〈0.05）,且与给药质量浓度呈正相关。结论：TSC能有效抑制SMMC-7721细胞增殖,其机制可能是通过上调Caspase-8、Caspase-9活性与p38MAPK、p53蛋白表达,最终激活Caspase-3,从而诱导细胞凋亡。
OBJECTIVE：To investigate the inhibitory effects of total saponins from Codonopsis pilosula（TSC）on human hepatocellular carcinoma SMMC-7721 cells and to explore their mechanism of action. METHODS：MTT method was used to determine cell viability after the TSC with concentrations of 0,100,200,400,600,800,1 000,1 200,1 400,1 600,1 800 and 2 000mg/L had acted on SMMC-7721 cells for 72 h,and the growth inhibition rates and median inhibitory concentration（IC50）were calculated. The flow cytometry was employed to determine the apoptosis after the TSC with the concentration of 911.0 mg/L had acted on SMMC-7721 cells for 24,48 and 72 h,and the apoptosis rates were calculated. The microplate reader and double-antibody sandwich enzyme-linked immunosorbent assay were adopted to determine the activities of Caspase-3,Caspase-8 and Caspase-9 as well as p38 MAPK and p53 protein expression after the TSC with concentrations of 0（blank control）,200,400,800 and 1 000 mg/L had acted on SMMC-7721 cells for 72 h. RESULTS：The TSC with concentrations of 100-2 000 mg/L could inhibit the proliferation of SMMC-7721 cells,and it had a positive correlation with the concentration;and the IC50 was 911.0 mg/L. The apoptosis rates were 21.10%,30.20% and 41.10% respectively after the TSC with the concentration of 911.0 mg/L had acted on cells for 24,48 and 72 h. Compared with blank control,the TSC with concentrations of 200,400,800 and 1 000 mg/L could enhance the protein expression of Caspase-3,Caspase-8,Caspase-9,p38 MAPK and p53 in cells,showing a positive correlation with the concentration.There was significant difference（P〈0.01 or P〈0.05）. CONCLUSIONS：TSC can effectively inhibit the proliferation of SMMC-7721 cells by the mechanism of increasing the protein expression of Caspase-8,Caspase-9,p38 MAPK and p53 to activate Caspase-3 and induce apoptosis.
出处
《中国药房》
CAS
北大核心
2015年第10期1356-1359,共4页
China Pharmacy
关键词
党参总皂苷
细胞凋亡
人肝癌SMMC-7721细胞
机制
Total saponins from Codonopsis pilosula
Apoptosis
Human hepatocellular carcinoma SMMC-7721 cells
Me-chanism
作者简介
中药师，硕士。研究方向：中药药物分析。电话：023-67063732。E—mail：fangzhe03@163．com
通信作者：副主任中药师，博士。研究方向：中药新药研究。电话：023—67063732。E-mail：ljlengjing@sina．com
