补肾疏肝方对肺腺癌A549细胞的抑制及凋亡相关因子研究  

Research about inhibition effects of Bushen Shugan Formula on human lung adenocarcinoma A549 cells and apoptosis related genes

作　　者：苗学红 王淑玲[2] 古兆森[1] 杜纪英[1] 

机构地区：[1]郑州大学第一附属医院中西医结合科,郑州450052 [2]郑州大学基础医学院,郑州450001

出　　处：《中华中医药杂志》2014年第8期2661-2664,共4页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：河南省科技厅科技攻关项目(No.112300410049)~~

摘　　要：目的:研究补肾疏肝方对肺腺癌A549细胞凋亡的影响及其凋亡相关基因的研究。方法:制备补肾疏肝方大鼠含药血清,与肺癌A549细胞共同培养,MTT法检测补肾疏肝方含药血清对A549增殖的影响。流式细胞术检测含药血清,对A549细胞周期的影响,Annexin V-FITC/PI双染流式细胞术观察含药血清对A549细胞凋亡的影响。Western blot检测含药血清对P53、Bcl-2蛋白表达的影响。结果:随着补肾疏肝方药物浓度的增加,处理时间的延长其抑制作用呈明显的时间-剂量效应,与顺铂联合作用效果最显著,其24、48、72h联合组的抑制率分别为26.71%、30.86%、41.74%;含药血清可以诱导细胞的凋亡,且随着浓度的增加G2/M期细胞的百分比增加,凋亡率与对照组比较差异有统计学意义(P<0.05);其在诱导细胞凋亡过程中,下调Bcl-2蛋白表达,也使P53蛋白表达量增高。结论:补肾疏肝方对A549细胞的增殖有抑制作用,并呈时间、剂量依赖方式,其作用机制可能是通过抑制Bcl-2蛋白表达,活化P53蛋白表达从而诱导肺癌细胞A549的凋亡而抑制细胞增殖。Objective： To study the apoptosis inducing effects and apoptosis related genes research of Bushen Shugan Formula on human lung adenocarcinoma A549 cells. Methods： Rat serum concentrating Bushen Shugan Formula was prepared and A549 cells were cultured in the drug serum with different concentrations. The proliferation of cells was detected by MTT assay. The flow cytometry was used to test the A549 cell cycle and cell apoptosis. Western-blot was used to test the expression of protein P53 and Bcl-2. Results： With the increase of Bushen Shugan Formula concentration and the extension of action time, the inhibitory effect on cells more obvious, and the effect of treatment group with DDP combined Bushen Shugan Formula was the most obvious. The inhibition rate of action time 24h, 48h and 72h was 26.71%, 30.86% and 41.74%, respectively. The drug serum could induce the cell apoptosis, and there were significant differences of apoptosis rate between the treatment group and the control group （P〈0.05）. With the increase of Bushen Shugan Formula concentration, the cell percentage in G2/M also increased. Drug could decrease the expression of Bcl-2, and increase the expression of P53. Conclusion： Bushen Shugan Formula could inhibit proliferation of A549 cells and demonstrate a dose and time dependent manner. The mechanism of inducing cell apoptosis and inhibiting cell proliferation with Bushen Shugan Formula might be that it could inhibit the Bcl-2 protein expression and promote P53 protein expression.

关 键 词：补肾疏肝方 肺癌 A549细胞 凋亡 凋亡相关基因 

分 类 号：R273[医药卫生—中西医结合]