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*

 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IC 50，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。

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

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 MTT assay. The A549 cells and A549/DDP cells were treated with the optimal medicated serum and cisplatin at different concentrations. The IC 50, resistance index 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 decoction decreases the resistance index by reducing the expression of PI3K in A549/DDP cells, thus increasing the sensitivity of A549/DDP cells to cisplatin.