姜黄素与博来霉素联用对A549细胞增殖抑制及细胞周期和凋亡的影响

Apoptosis Inducing and Antiproliferation Effects of Curcumin Combined with Bleomycin on the Human Lung Cancer Cell Line A549

目的:研究姜黄素和博来霉素联合应用对人肺泡Ⅱ型样细胞A549细胞增殖和细胞周期及细胞凋亡的影响,探讨姜黄素抗博来霉素诱导的肺纤维化机制。方法:采用MTT法观察姜黄素和博来霉素对A549细胞的增殖抑制作用,并用流式检测细胞周期分布,凋亡试剂盒检测细胞凋亡。结果:姜黄素及博来霉素能抑制A549细胞的增殖,并呈浓度及时间依赖性。姜黄素(Cur)浓度为60μmol/L与博来霉素(BLM)2、4、6、8μg/mL联合24h的抑制率显著高于单用博来霉素组;姜黄素可引起体外培养的A549细胞G0/G1期阻滞;而且对G0/G1期阻滞随作用时间延长而增强。单用博来霉素刺激细胞,培养24h和48h,对细胞周期各个时期的影响没有统计学意义;姜黄素联合博来霉素诱导A549细胞凋亡,细胞核变的不规则皱缩,可见凋亡小体。结论:姜黄素联合博来霉素在体外对A549细胞的增殖有明显的抑制作用,对细胞周期G0/G1期有显著阻滞作用,并可改变细胞凋亡;姜黄素和博来霉素联合应用可能通过改变肺泡上皮细胞细胞周期分布,并通过影响肺泡细胞凋亡从而起到抑制博来霉素治疗过程中肺纤维化的发生发展。

Objective：To investigate the apoptosis inducing and antiproliferation effects of curcumin combined with bleomycin on the human type II alveolar epithelial cell line A549 in vitro.Methods：MTT was used to measure inhibitory effects of curcumin combined with bleomycin on growth of A549 cells,flow cytometry were used to analyze the cell cycle,TUNEL system and fluorescence microscope were used to observe the morphology of apoptotic cells.Results：Curcumin and bleomycin inhibited the growth of A549 in a concentration and time dependent manner,Compared with either curcumin or bleomycin alone,combining curcumin at 60μmol/L,with bleomycin at 2,4,6,8μg/mL respectively increased the growth inhibition rate of A549 cells（P0.05） significantly;curcumin induce G0/G1 inhibition or arrest in a time dependent manner,curcumin combined with bleomycin also induced apoptosis,nuclear and cytoplasmic shrinkage,chromatin condensation and apoptotic bodies can been seen.Conclusion：Curcumin combined with bleomycin inhibited the growth of A549 significantly and also induced A549 apoptosis,but only curcumin induce G0/G1 inhibition or arrest.Through changing the cell cycle phase and the apoptosis of alveolar epithelial cell,curcumin may inhibit lung fibrosis during the development after the treatment of bleomycin.