补中益气汤含药血清同siRNA对A549/DDP细胞MRP表达影响的比较

To Compare Effect of Buzhong Yiqi Decoction Serum with siRNA on MRP Expression of A549/DDP Cells

摘要:

目的：观察补中益气汤含药血清对人肺腺癌耐顺铂药细胞株（A549/DDP）多药耐药相关蛋白（multidrug resistance-associated protein，MRP）表达的影响，并将结果同小分子干扰RNA（small interfering RNA，siRNA）技术特异性沉默MRP基因相比较。方法：采用血清药理学方法制备含药血清，随机对照法将18只SD大鼠分为空白对照组（给生理盐水10 mL·kg^-1），补中益气汤高、低剂量组（11.34，2.84 g·kg^-1，10 mL·kg^-1），每日ig 给药1 次，连续3 d后制备补中益气汤药含药血清，使用siRNA转染A549/DDP细胞，取处于对数生长期细胞于6 孔板中，每孔加40 μL质粒（0.1 g·L^-1），24 h后，利用RT-PCR（reverse transcription-polymerase chain reaction，RT-PCR）技术检测MRP mRNA的表达和蛋白质印迹法（Western blot）技术检测MRP蛋白表达。实验分组为：空白对照组、补中益气汤含药血清低、高剂量处理组和MRP siRNA处理组。结果：与空白对照组相比，补中益气汤含药血清低、高剂量处理组和MRP siRNA处理组的MRP mRNA及蛋白表达均减少，具有统计学意义（P<0.05）。结论：补中益气汤含药血清能降低A549/DDP细胞上的MRP蛋白表达，且同特异性沉默MRP基因后效果类似。

Abstract:

Objective: To compare the effect of Buzhong Yiqi decoction serum with small interfering RNA (siRNA)on the multidrug resistance-associated protein(MRP)expression of A549/DDP lung drug resistance cells. Method: Buzhong Yiqi decoction serum was prepared. 18 rats were divided into the blank group (normal saline, 10 mL·kg^-1), high (11.34 g·kg^-1) and low (2.84 g·kg^-1) dose groups of Buzhong Yiqi decoction. At the end of the 3th day serum was obtained. siRNA was used to silence MRP gene into A549/DDP cells. Reverse transcription-polymerase chain reaction(RT-PCR) and Western blot analyses were performed for MRP mRNA, and MRP expression in A549/DDP cells. Cells were randomized into control groups;Buzhong Yiqi decoction lower does groups;Buzhong Yiqi decoction higher does groups, and MRP siRNA groups. Result: This study showed that three treatment groups could significantly decreased the MRP mRNA expression, and the expression of MRP at the level of protein(P<0.05). Conclusion: These results suggest the Buzhong Yiqi decoction serum could decrease the expression of MRP in A549/DDP cells, and has similar effect with siRNA-mediated specific gene silencing in A549/DDP cells.