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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40μL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。
Objective: To observe the effect of Buzhong Yiqi decoction-containing serum on the expression of multidrug resistance-associated protein (MRP) in human lung adenocarcinoma A549 / DDP cell line, Small interfering RNA (siRNA) technology specifically silences MRP genes. Methods: Serum pharmacological methods were used to prepare the serum containing drugs. 18 SD rats were randomly divided into blank control group (given normal saline 10 mL · kg -1), Buzhong Yiqi Tang high and low dose group (11.34 , 2.84 g · kg-1 and 10 mL · kg-1 respectively). The rats were given ig once a day for 3 days. The serum of Buzhong Yiqi decoction was prepared. A549 / DDP cells were transfected with siRNA. The cells were counted in 6-well plates with 40 μL plasmid (0.1 g · L-1) for 24 h. The expression of MRP mRNA was detected by reverse transcription-polymerase chain reaction (RT-PCR) And Western blotting were used to detect MRP protein expression. The experimental groups were as follows: blank control group, Buzhong Yiqi Decoction containing serum low, high dose treatment group and MRP siRNA treatment group. Results: Compared with the blank control group, MRP mRNA and protein expression of Buzhong Yiqi Decoction containing low serum, high dose and MRP siRNA groups decreased, with statistical significance (P <0.05). Conclusion: Buzhong Yiqi decoction containing serum can reduce the expression of MRP protein in A549 / DDP cells, which is similar to the effect of MRP gene silencing.