益肺解毒方逆转A549/DDP耐药及诱导其凋亡的实验研究

Reverse Effect of Yifei Jiedu Decoction on Cisplatin Resistant Human Lung Adenocarcinoma Cell Line A549/DDP and the Mechanism of Induce Cell Apoptosis

摘要: 目的：探讨益肺解毒方(YFJDF)含药血清逆转人肺腺癌A549/DDP细胞耐药性及诱导其凋亡的机制。方法：制备YFJDF含药血清，CCK8法分别检测不同浓度的顺铂对A549、A549/DDP细胞的增殖抑制率，不同浓度YFJDF对A549、A549/DDP细胞的增殖抑制率，YFJDF联合各浓度顺铂对A549/DDP细胞的增殖抑制率，并根据结果计算IC50值、A549/DDP细胞对顺铂的耐药指数及YFJDF逆转A549/DDP细胞的逆转耐药倍数。流式细胞术检测各组细胞凋亡率、West-blot检测各组细胞Bax、Bcl-2蛋白的表达水平。结果：不同浓度的DDP作用A549、A549/DDP细胞48 h后，其IC50值分别为4.49 μg/mL和45.09 μg/mL，计算出的耐药指数为10.05倍。不同浓度YFJDF对A549/DDP细胞的增殖有抑制作用，并且随着YFJDF浓度的升高和药物作用时间的延长对A549/DDP细胞的抑制作用也逐渐增加，选择中剂量的YFJDF、并作用48 h用于后续逆转实验。YFJDF联合各浓度顺铂对A549/DDP细胞的抑制作用更为明显，其48 h的IC50值为29.37 μg/mL，逆转倍数为1.53倍。流式细胞仪检测结果显示：YFJDF + DDP组细胞凋亡率显著增加，可达到69.3%；Western blot结果显示：YFJDF + DDP组Bax的表达量显著增加、Bcl-2的表达量明显减少。结论：益肺解毒方能逆转A549/DDP细胞对顺铂的耐药，其机制可能是促进顺铂诱导A549/DDP凋亡。

Abstract: Objective: To study the reversal effect of Yifei Jiedu Decoction on cisplatin resistant human lung adenocarcinoma cell line A549/DDP and to explore the mechanism of induce cell apoptosis. Methods: Prepare YFJDF medicated serum. The method of CCK8 was used to detect the proliferation inhibition rate of different concentrations of DDP on A549 and A549/DDP cells, the proliferation inhibition rate of different concentrations of YFJDF on A549 and A549/DDP cells and YFJDF combined with different concentrations of DDP on A549/DDP cells. IC50, the drug resistance index of A549/DDP cells to DDP and the reversal drug resistance fold of YFJDF were calculated according to the results. Apoptosis rate in each group detected by flow cytometry. The expression level of apoptosis-related protein Bax and Bcl-2 was detected by western blot in each group. Results: After different concentrations of DDP acted on A549 and A549/DDP cells for 48 hours, the IC50 was 4.49 μg/mL and 45.09 μg/mL, drug resistance index was 10.05 times. Different concentrations of YFJDF have an inhibitory effect on the proliferation of A549/DDP cells, and with the increase of YFJDF concentration and the prolongation of drug action time, the inhibitory effect on A549/DDP cells will gradually increase. A medium dose of YFJDF was selected and the action time of 48 hours was selected for the subsequent reversal experiment. The inhibitory effect of YFJDF combined with DDP at various concentrations on A549/DDP cells was more obvious, and its 48 h IC50 was 29.37 μg/mL, and the reversal drug resistance fold was 1.53 times. The results of flow cytometry showed that the apoptosis rate of the YFJDF + DDP group increased significantly, reaching 69.3%; Western blot results showed that the expression of Bax in the YFJDF + DDP group increased significantly, and the expression of Bcl-2 decreased significantly. Conclusion: Yifei Jiedu Decoction can reverse the resistance of A549/DDP cells to DDP, and its mechanism may be to promote the apoptosis of A549/DDP induced by DDP.