爱康方含药大鼠血清对人肺腺癌A549细胞内Bad、Caspase-9蛋白及mRNA的作用  

Effect of serum containing Aikang Formula on Bad,Caspase-9 protein and mRNA in human lung adenocarcinoma A549 cells LIU

作　　者：刘文杰 李巧玲[1] 马科[1] LIU Wen-jie;LI Qiao-ling;MA Ke(Ningxia Medical University,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学,750004

出　　处：《中国实用医药》2022年第13期191-196,共6页China Practical Medicine

基　　金：国家自然科学基金资助项目(项目编号:81660829,81960860)。

摘　　要：目的 探讨爱康方含药血清对人肺腺癌A549细胞内B淋巴细胞瘤-2基因相关启动子(Bad)、Caspase-9蛋白(Caspase-9蛋白)及mRNA的表达作用。方法 40只大鼠,采用随机分组法分为空白对照组(NS)、环磷酰胺组(CTX)、爱康方组(AKF)和爱康方联合环磷酰胺组(AC),每组10只。以人肺腺癌A549细胞为研究对象,依据前期研究结果选定20%的含药血清为最佳干预浓度,用各组含药血清分别干预细胞48、72 h后收集细胞用于检测。通过蛋白质印迹法(Western blot)测定干预后目的细胞中Bad、Caspase-9蛋白表达,实时荧光定量PCR技术(RT-qPCR)法测定Bad、Caspase-9 mRNA的表达。比较各组不同方法检测的Bad mRNA、Caspase-9 mRNA表达水平及Bad、Caspase-9蛋白表达水平。结果 48、72 h时,环磷酰胺组、爱康方组和爱康方联合环磷酰胺组Bad mRNA表达水平高于空白对照组,爱康方联合环磷酰胺组Bad mRNA表达水平高于环磷酰胺组和爱康方组,差异具有统计学意义(P<0.05);48 h时,环磷酰胺组Bad mRNA表达水平高于爱康方组,差异具有统计学意义(P<0.05)。72 h时,爱康方组和爱康方联合环磷酰胺组Bad mRNA表达水平高于本组48 h,差异具有统计学意义(P<0.05)。48、72 h时,环磷酰胺组、爱康方组和爱康方联合环磷酰胺组Caspase-9 mRNA表达水平高于空白对照组,爱康方联合环磷酰胺组Caspase-9 mRNA表达水平高于环磷酰胺组和爱康方组,差异具有统计学意义(P<0.05);48 h时,环磷酰胺组Caspase-9 mRNA表达水平高于爱康方组,差异具有统计学意义(P<0.05)。72 h时,爱康方组和爱康方联合环磷酰胺组Caspase-9 mRNA表达水平高于本组48 h,差异具有统计学意义(P<0.05)。48、72 h时,环磷酰胺组、爱康方组和爱康方联合环磷酰胺组Bad蛋白表达水平明显高于空白对照组,环磷酰胺组、爱康方联合环磷酰胺组高于爱康方组,爱康方联合环磷酰胺组高于环磷酰胺组,差异具有统计学意义(P<0.05)。Objective To discuss the effect of serum containing Aikang Formula on Bcl-xl/Bcl-2 associated death promoter(Bad),Caspase-9 protein and mRNA in human lung adenocarcinoma A549 cells.Methods 40 rats were randomly divided into blank control group(NS),cyclophosphamide group(CTX),AKF group(AKF)and AKF combined with cyclophosphamide group(AC),with 10 rats in each group.Taking human lung adenocarcinoma A549 cells as the research object,and 20%drug-containing serum was selected as the optimal intervention concentration based on the results of the previous study.The cells were collected after 48 and 72 h of intervention with each group of drug-containing serum for assay.The expression of Bad and Caspase-9 protein in the target cells after the intervention was measured by Western blot,and the expression of Bad and Caspase-9 mRNA was measured by real time quantitative polymerase chain reaction(RT-qPCR).The expression levels of Bad mRNA and Caspase-9 mRNA and the protein expression levels of Bad and Caspase-9 detected by different methods in each group were compared.Results At 48 and 72 h,the expression levels of Bad mRNA in the cyclophosphamide group,the AKF group and the AKF combined with cyclophosphamide group were higher than those in the blank control group,and the expression level of Bad mRNA in AKF combined with cyclophosphamide group was higher than that in cyclophosphamide group and AKF group.All the differences were statistically significant(P<0.05).At 48 h,the expression level of Bad mRNA in the cyclophosphamide group was higher than that of the AKF group,and the difference was statistically significant(P<0.05).At 72 h,the expression levels of Bad mRNA in the AKF group and the AKF combined with cyclophosphamide group were higher than that of this group at 48 h,and the difference was statistically significant(P<0.05).At 48 and 72 h,the expression levels of Caspase-9 mRNA in the cyclophosphamide group,the AKF group and the AKF combined with cyclophosphamide group were higher than those in the blank control group,and

关 键 词：人肺腺癌 A549细胞 爱康方 B淋巴细胞瘤-2基因相关启动子 CASPASE-9蛋白 

分 类 号：R285.5[医药卫生—中药学]