温下方含药血清诱导A549/DDP细胞凋亡及对Bcl-2,Bax,p53蛋白表达的影响

目的:研究温下方含药血清诱导人肺腺癌耐药细胞株A549/DDP凋亡及对Bcl-2,Bax,p53蛋白表达的影响。方法:制备温下方含药血清,高效液相色谱法(HPLC)定性检测含药血清中人参皂苷Rb1、大黄素和乌头碱。A549/DDP细胞分为DDP干预组,含药血清+DDP干预组,正常血清+DDP干预组。孵育24 h后,流式细胞仪检测细胞凋亡率,激光共聚焦显微镜检测Bcl-2,Bax及p53蛋白表达。结果:HPLC结果显示含药血清中含有人参皂苷Rb1、大黄素及乌头碱。含药血清作用后,与DDP干预组比较,含药血清+DDP干预组细胞凋亡率增高至(21.82±3.27)%(P<0.05);细胞内Bcl-2蛋白表达降低至2.81±0.02(P<0.05);Bax蛋白表达增高至2.90±0.08(P<0.05);p53蛋白表达增高至3.58±0.09(P<0.05)。结论:温下方含药血清明显诱导A549/DDP细胞凋亡,显著降低Bcl-2蛋白表达,提高Bax,p53蛋白表达,可能从诱导耐药细胞凋亡角度逆转耐药。

Effect of Wenxia Recipe Medicated Serum on Inducing Apoptosis ofA549/DDP Cells and Expression of Bcl-2,Bax and p53

Objective: To investigate the effect of Wenxia recipe medicated serum on inducing apoptosis of A549/DDP cells and expression of Bcl-2,Bax and p53 protein by using serum pharmacological methods in vitro. Method: Drug-containing serum was prepared and analyzed by HPLC. Furthermore,A549/DDP cells were divided into DDP group,drug-containing serum + DDP group and normal serum +DDP group.After incubated 24h,cell apoptosis was evaluated by laser flow cytometry and the expression of Bcl-2,Bax and p53 protein in cells was investigated by laser scanning confocal microscope. Result: There were ginsenoside Rb₁,emodin,aconitine in drug-containing serum. Compared to DDP group,the apoptosis rate of drug-containing serum + DDP group rose to (21.82±3.27)% (P<0.05),the expression of Bcl-2 significantly dropped to 2.81±0.02 (P<0.05) and the expression of Bax and p53 significantly rose to 2.90±0.08 and 3.58±0.09 (P<0.05) respectively. Conclusion: Wenxia recipe medicated serum could significantly induce apoptosis of A549/DDP,decrease the expression of Bcl-2 and increase the expression of Bax and p53.This study showed that Wenxia recipe might reverse the mutidrug-resistance of A549/DDP cell by inducing apoptosis.