艾烟可吸入颗粒物对人肺腺癌A549细胞凋亡的影响

Effect of PM10 in Moxa Smoke on Apoptosis of Human Lung Adenocarcinoma Cell Line-A549 Cells

目的观察艾烟可吸入颗粒物(PM10)对人肺腺癌A549细胞凋亡的影响,并探讨其诱导凋亡的可能机制。方法采用体外实验方法,以A549细胞为研究对象,采用Hoechest33258染色法,荧光显微镜观察艾烟PM10对A549细胞凋亡的影响,检测细胞内Ca2+水平、核因子(NF)-κB p65表达量及细胞内活性氧(ROS)的水平。结果艾烟PM10处理4 h在400μg/mL时出现部分凋亡细胞。艾烟PM10干预A549细胞4 h后,随着浓度增加,细胞内Ca2+水平显著增加(P<0.01)。与空白对照组比较,艾烟干预4 h,各浓度组A549细胞内NF-κB p65表达量均降低(P<0.05);干预20 h,70μg/mL组和280μg/mL组NF-κB p65表达量均降低(P<0.05),140μg/mL组NF-κB p65含量无明显变化(P>0.05)。相同浓度下,不同时点艾烟干预各组间比较,NF-κB p65表达量无明显变化(P>0.05)。与空白对照组比较,艾烟PM10处理各组细胞内ROS水平均显著降低(P<0.05)。结论艾烟PM10能诱导A549细胞凋亡、提高A549细胞内Ca2+水平。

Objective To observe the effect of PM10 （inhalable particles） in moxa smoke on apoptosis of human lung adenocarcinoma cell line-A549 cells, and explore the possible mechanisms of inducing apoptosisMethods The A549 cells were studied in vitro experiment method, which were stained by Hoechest33258 staining method. Their morphological changes of apoptosis were observed by the fluorescence microscopy. The levels of intracellular Ca2＋ and reactive oxygen species （ROS）, and expression of NF-κB p65 were also measured.Results Some apoptotic cells were observed after treated with moxa smoke PM10 in the concerntration of 400μg/mL. After 4 hours intervention by moxa smoke PM10 in A549 cells, the intracellular Ca2＋ level increased significantly （P〈0.01）. Compared with the blank control group, the expression of NF-κB p65 decreased significantly （P〈0.05） after intervention of moxa smoke PM10 with different concerntrations for 4 hours. When the A549 cells were cultured with moxa smoke PM10 for 20 hours, the expression of p65 decreased significantly （P〈0.05） in the concerntrations of 70, 280μg/mL, while there was no significant change in the concerntration of 140μg/mL （P〉0.05）. Compared with the blank control group, ROS level was significantly lower （P〈0.05） in A549 cells after intervention of moxa smoke PM10.Conclusion PM10 in moxa smoke could induce apoptosis of A549 cells, could increase cytosolic Ca2＋ level.