补中益气汤含药血清对人肺腺癌A549/DDP细胞耐药作用的影响  

Effects of Buzhong Yiqi decoction-medicated serum on drug resistance of human lung adenocarcinoma cell line A549/DDP to cisplatin

作　　者：刘亚莉[1,2] 王莹[1] 易佳丽[1] 史妍婷[1] 刘春英[1] 

机构地区：[1]辽宁中医药大学基础医学院病理教研室,辽宁沈阳110032 [2]辽宁卫生职业技术学院医学技术系卫生检验教研室,辽宁沈阳110101

出　　处：《中国病理生理杂志》2014年第2期233-238,共6页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81072743);辽宁省教育厅资助项目(No.L2010355)

摘　　要：目的:观察补中益气汤含药血清对人肺腺癌细胞株A549/DDP耐药作用的影响并探讨其机制。方法:采用血清药理学方法制备含药血清;用MTT比色法筛选出最佳含药血清;以最佳含药血清和不同浓度梯度的顺铂干预A549和A549/DDP细胞,MTT比色法计算2种细胞的IC50,同时计算A549/DDP耐药指数和逆转指数;将A549/DDP细胞分为对照血清组、最佳含药血清组、LY294002组、LY294002+最佳含药血清组(联合组)和阴性对照组,给予相应的处置因素后,培养48 h,采用免疫细胞化学方法、Western blotting和real-time PCR等方法检测各组中PI3K、Akt蛋白与mRNA的表达情况。结果:10%中剂量含药血清作用48 h为最佳含药血清及最佳作用时间。最佳含药血清与顺铂联合作用时,A549/DDP细胞对顺铂的敏感性明显增强,最佳含药血清的逆转指数约为2.46。最佳含药血清组和联合组的PI3K、Akt蛋白和mRNA的表达水平均下降,与对照血清组比较差异有统计学意义(P<0.05)。LY294002组的PI3K、Akt蛋白和mRNA的表达水平与对照血清组比较,差异无统计学意义(P>0.05)。结论:补中益气汤通过减少PI3K的表达而降低A549/DDP细胞的耐药指数,增加A549/DDP细胞对顺铂的敏感性,干预或调整A549/DDP细胞对顺铂的耐药作用。AIM： To investigate the effects of Buzhong Yiqi decoction-medicated serum on the ;drug resistance of human lung adenocarcinoma cell line A549/DDP to cisplatin. METHODS ： Medicated serum containing Buzhong Yiqi decoction was prepared. The optimal dose of the medicated serum was selected by MTr assay. The A549 cells and A549/ DDP cells were treated with the optimal medicated serum and cisplatin at different concentrations. The IC50, resistance in- dex and reversal index were determined. The cells were divided into control serum group, optimal medicated serum group, LY294002 group, LY294002 ＋ optimal medicated serum group （combination group） and negative group. The expression of PI3K and Akt at mRNA and protein levels was detected by the methods of immunocytochemistry, Western blotting and real-time PCR. RESULTS ： Treatment with 10% middle dose of medicated serum for 48 h was the optimal dose and time for medicated serum. The sensitivity of A549/DDP cells to cisplatin was obviously enhanced when the cells were exposed to the optimal medicated serum with the reversal index of 2.46. The expression of PI3K and Akt at mRNA and protein levels in optimal medicated serum group and combination group was significantly decreased. CONCLUSION： Buzhong Yiqi de- coction decreases the resistance index by reducing the expression of PI3K in A549/DDP cells, thus increasing the sensitivi- ty of A549/DDP cells to cisplatin.

关 键 词：补中益气汤 含药血清 肺腺癌 耐药性 PI3K蛋白 AKT蛋白 

分 类 号：R273[医药卫生—中西医结合]