补中益气汤含药血清对A549/DDP中p-Akt和P-gp表达的影响  

Effect of Serum Containing Buzhong Yiqi Decoction on p-Akt and P-gp of Human Lung Adenocarcinoma Cells A549 /DDP

作　　者：易佳丽[1] 井欢[1] 王莹[1] 刘亚莉[1] 刘春英[1] 

机构地区：[1]辽宁中医药大学,辽宁沈阳110847

出　　处：《中国实验方剂学杂志》2013年第17期229-234,共6页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81072743);辽宁省教育厅项目(L2010355)

摘　　要：目的:研究补中益气汤含药血清逆转人肺腺癌顺铂耐药细胞株(A549/DDP)耐顺铂(DDP)作用及其对磷酸化丝氨酸/苏氨酸蛋白激酶(p-Akt)和P-糖蛋白(P-gp)表达的影响。方法:采用血清药理学方法制备含药血清,随机对照法将24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、中、低剂量组(11.34,5.67,2.84 g·kg-1,10 mL·kg-1),每日ig给药1次,连续7 d后提取血清。将A549/DDP分为空白血清组,补中益气汤高、中、低剂量含药血清组,阻滞剂组,高剂量含药血清联合阻滞剂(LY294002)组,MTT法检测各组细胞对DDP的IC50,并计算耐药逆转倍数。免疫细胞化学法检测各组细胞中p-Akt和P-gp蛋白的表达;逆转录-多聚合酶链式反应(RT-PCR)法检测各组细胞中Akt和P-gp的mRNA表达。结果:与空白血清组比较,补中益气汤中剂量以上的含药血清可明显降低A549/DDP对DDP的IC50(P<0.05),并呈浓度依赖效应,与阻滞剂联合效果最佳(P<0.05);与A549/DDP空白血清组相比,补中益气汤中、高剂量含药血清组,阻滞剂组和高剂量联合阻滞剂组的p-Akt和P-gp的表达显著下降(P<0.01,P<0.05),其中以高剂量联合阻滞剂组下降最多(P<0.01);与A549/DDP空白血清组相比,中、高剂量含药血清组,阻滞剂组和高剂量联合阻滞剂组的P-gp的mRNA表达量都显著减少(P<0.01,P<0.05),中、高剂量含药血清组,高剂量联合阻滞剂组的p-Akt的mRNA表达量也显著减少(P<0.05)。结论:补中益气汤含药血清可逆转A549/DDP细胞的顺铂耐药,与LY294002联合应用效果更佳,其逆转机制可能与减少细胞内p-Akt和P-gp的表达,增强A549/DDP对DDP的敏感性有关。Objective：To analyses the reversal effect of serum containing Buzhong Yiqi decoction on multidrug resistance of human adenocarcinoma of lung cell line A549/DDP,and observe the impacts on expression of phosphorylation of senine threonine protein kindse（p-Akt） and P-glycoprotein（P-gp）.Method：The serum containing Buzhong Yiqi decoction was prepared by adopting the method of serum pharmacology.24 rats were divided into the blank group（normal saline,10 mL·kg-1）,high（1.314 g·kg-1）,middle（5.76 g·kg-1） and low（2.84 g·kg-1） dose groupsof Buzhong Yiqi decoction.At the end of the 7th day serum was obtained.A549/DDP was divided into blank serum group,high,middle and low dose groups of serum drug-containing.LY294002 group,high dose combined with LY294002 group,IC50 was observed by MTT assay,and the reversal fold was computed.Immunohistochemical（IHC） was used to assay the protein expression of p-Akt and P-gp,RT-PCR was used determine to the content of mRNA of Akt and P-gp.Result：Compared with blank serum group,IC50 in middle and high dose group was decreased（P ＆lt; 0.05）,and it was observed to exert a concentration-dependent inhibition on the proliferation of A549/DDP cell line,the largest decline was in high dosage combined with LY294002 group（P ＆lt; 0.01）.The protein expression of p-Akt and P-gp was significantly decreased in middle and high group,LY294002 group combined with high dose LY294002 group（P ＆lt; 0.01,P ＆lt; 0.05）.The mRNA expression of P-gp and Akt was significantly decreased in middle and high dose group,LY294002 group combined with high dose group（P ＆lt; 0.01,P ＆lt; 0.05）.Conclusion：The serum containing Buzhong Yiqi decoction could moderately reverse the multidrug resistance of A549/DDP cells through reducing the expression of p-Akt and P-gp and increasing the sensitivity of 549/DDP to DDP.

关 键 词：A549 DDP细胞株 磷酸化丝氨酸 苏氨酸蛋白激酶 P-糖蛋白 补中益气汤 多药耐药 

分 类 号：R285.5[医药卫生—中药学]