补中益气汤含药血清逆转人肺腺癌耐药细胞株A549/DDP耐顺铂作用及对Survivin表达的影响

目的研究补中益气汤含药血清逆转人肺腺癌耐药细胞株A549/DDP耐顺铂作用及对Survivin表达的影响。方法采用随机对照法将24只SD大鼠分为高、中、低剂量组和空白对照组,制备补中益气汤高剂量（补中益气汤1.134 g/mL,2 mL）、中剂量（补中益气汤0.576 g/mL,2 mL）、低剂量（补中益气汤0.284 g/mL,2 mL）含药血清,采用MTT法检测不同剂量和浓度的补中益气汤含药血清对A549及A549/DDP细胞的增殖抑制效应,计算耐药逆转倍数;采用免疫细胞化学法、Western blot技术、免疫荧光三种方法检测Survivin在两细胞株中的表达。结果补中益气汤含药血清对A549和A549/DDP细胞有明显抑制作用,10%各剂量组抑制率均高于5%相应剂量组,且随着剂量浓度的升高,抑制率也逐渐升高;与10%空白组比较,10%中、低剂量组抑制率升高（P〈0.05）;10%中剂量含药血清作用后,A549和A549/DDP IC50均降低（P〈0.05）,逆转倍数均升高,对A549/DDP细胞的逆转倍数达到2.46,使A549/DDP对顺铂的耐药性下降;与本组A549比较,免疫细胞化学法、Western blot法、免疫荧光法均检测到A549/DDP细胞中Survivin的表达量均降低（P〈0.05）,与10%空白组比较,10%中剂量组A549/DDP均降低（P〈0.05）。结论 10%中剂量补中益气汤含药血清对A549及A549/DDP细胞都有抑制作用,并可逆转A549/DDP细胞的顺铂耐药,其逆转机制可能与减少细胞内Survivin的表达,增强A549/DDP对化疗药物的敏感性有关。

Buzhong Yiqi Decoction Containing Serum Reversed Resistance of A549/DDP to Cisplatin and Its Effect on the Expression of Survivin. an Experimental Research

Objective To analyze the reversal effect of Buzhong Yiqi Decoction （BYD） on multi- drug resistance of human adenocarcinoma of lung cell line A549/DDP, and to study its effect on the expression of survivin by using serum pharmacological methods in vitro. Methods Totally 24 SD rats were divided into the high, medium and low dose groups, and the blank control group by randomized controlled method. The high dose BYD containing serum （1. 134 g/mL, 2 mL）, the middle dose BYD containing serum （0. 576 g/mL, 2 mL）, and the low dose BYD containing serum （0. 284 g/mL, 2 mL） were prepared. The inhibitory effects of different dose and concentrations BYD on the proliferation of A549 and A549/DDP cells were detected by MTT assay, and the drug resistance reversal fold was calculated. The expression of Survivin in the two cell strains were detected respectively by immunohistochemical assay, Western blot, and immunofluorescence method. Results BYD containing serum showed obvious inhibitory effect on the growth of A549 and 549/DDP. The inhibition rates of 10% dose groups were higher than those of 5% dose groups. Besides, it gradually increased along with increased concentrations. Compared with 10% blank control group, the inhibition rate increased in 10% middle and low dose groups （P 〈0.05）. Af-ter acted with 10% middle dose BYD containing serum, IC50 of A549 and A549/DDP were both reduced （P 〈0.05）, reversal fold （RF） both increased. Its reversal ratio on A549/DDP cells was 2.46, decreasing the resistance of A549/DDP to DDP. Compared with A549 in the same group, the expression of Survivin was detected to decrease by immunocytochemical assay, Western blot, and immunofluorescence method （P 〈0.05）. Compared with 10% blank control group, the inhibition rate decreased in 10% middle dose group （P 〈0.05）. Conclusions 10% middle dose BYD containing serum could significantly inhibit the apoptosis of A549 and A549/DDP. Besides, it could moderately reverse the multidrug resistance of A549/ DDP cell