[关键词]
[摘要]
目的 观察注射用益气复脉(冻干)(YQFM)与顺铂(Cisplatin,DDP)联合对肺癌细胞株A549增殖抑制的协同效应及作用机制。方法 采用CCK-8法检测YQFM、DDP以及两药联用对肺癌细胞株A549的增殖影响,并应用两药相互作用指数评价药物的联合效应;采用荧光探针JC-1来检测线粒体膜电位的变化,Wester Blotting检测凋亡相关蛋白Bax、Cleaved-Caspase 3表达。结果 药物作用时间48 h,与DDP等剂量组比较,联合用药(YQFM 16.25 mg/mL+DDP 1.25 μg/mL、YQFM 16.25 mg/mL+DDP 2.5 μg/mL、YQFM 16.25 mg/mL+DDP 5.00 μg/mL)组细胞增殖抑制率显著增加(P<0.05)。与对照组比较,各给药组凋亡率均显著升高(P<0.05);与DDP组比较,联合用药组凋亡率显著升高(P<0.05)。与对照组比较,DDP 5.0、10.0 μg/mL组和各联合用药组促凋亡蛋白Bax表达显著增加(P<0.05);DDP质量浓度为5.0、10.0 μg/mL时,联合用药组与DDP单独给药组比较,Bax蛋白表达量显著升高(P<0.05)。与对照组比较,各给药组凋亡蛋白Cleaved-caspase 3表达量显著升高(P<0.05);DDP质量浓度为5.0、10.0 μg/mL时,联合用药组与DDP单独给药组比较,Cleaved-caspase 3蛋白表达量显著升高(P<0.05)。结论 YQFM联合DDP对A549细胞增殖存在协同抑制作用,机制可能与诱导线粒体凋亡相关。
[Key word]
[Abstract]
objective To observe the synergistic effect and mechanism of combined injection of Yiqi Fumai Lyophilized Injection (YQFM) and Cisplatin (DDP) on the proliferation inhibition of A549 lung cancer cell line. Methods The CCK-8 method was used to detect the effect of single DDP and YQFM, and combination of two drugs on the proliferation of lung cancer cell line A549. The changes of mitochondrial membrane potential were detected by fluorescent probe JC-1, which labeled the mitochondrial membrane potential. Western Blot was used to detect the expression of apoptosis-related proteins Bax and Caspase3. Results Compared with DDP equal dose group, the inhibition rate of cell proliferation was significantly increased in YQFM 16.25 mg/mL + DDP 1.25 μg/mL, YQFM 16.25 mg/mL + DDP 2.5 μg/mL, YQFM 16.25 mg/mL + DDP 5.00 μg/mL groups (P<0.05). Compared with the control group, the apoptotic rate of each drug group was significantly higher (P<0.05), and the apoptotic rate of the combined drug group was significantly higher (P<0.05) than that of the DDP group. Compared with control group, the expression of apoptotic protein Bax in DDP 5.0, 10.0 μg/mL and combination groups was significantly higher (P<0.05); when DDP concentration was 5.0 and 10.0 μg/mL, the expression of Bax protein in combination group was significantly higher than that in DDP alone group (P<0.05). Compared with the control group, the expression of apoptotic protein Cleaved-caspase 3 was significantly increased in each group (P<0.05). When the concentration of DDP was 5.0 and 10.0 μg/mL, the expression of Cleaved-caspase 3 in the combined group was significantly higher than that in the DDP alone group (P<0.05). Conclusion YQFM combined with DDP could inhibit the proliferation of A549 cells synergistically, and the mechanism might be related to the induction of mitochondrial apoptosis.
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[基金项目]
天津市中药注射剂关键技术校企协同创新实验室建设项目(17PTSYJC00090)