丹皮酚对脂多糖刺激气道上皮细胞的影响

Effects of paeonol on airway epithelial cells stimulated by lipopolysaccharide

[目的]探讨丹皮酚对脂多糖(LPS)所刺激气道上皮细胞的影响及其作用机制.[方法]采用流式细胞术检测以15、30μmol/L丹皮酚预处理经LPS刺激的16HBE细胞ROS的表达水平及细胞凋亡情况,采用Western blot法检测细胞色素C(Cyt-C)、caspase-9、caspase-3、Bax及Bcl-2的表达水平.[结果]与LPS组比较,丹皮酚低剂量和高剂量组LPS刺激16HBE细胞中ROS的表达均显著降低(P<0.05)、细胞早期和晚期凋亡率(P<0.05)及细胞胞浆中Cyt-C蛋白表达水平均显著降低(P<0.01),线粒体中Cyt-C蛋白表达水平显著升高(P<0.01);丹皮酚显著降低LPS刺激16HBE细胞中caspase-9、caspase-3的表达(P<0.01)及促凋亡蛋白Bax的表达(P<0.01),明显升高抗凋亡蛋白Bcl-2的表达(P<0.05).[结论]丹皮酚通过抑制线粒体凋亡通路减轻LPS刺激对气道上皮细胞的影响.

OBJECTIVE To explore the effects of paeonol on airway epithelial cells stimulated lipopolysaccharide(LPS)and its mechanism.METHODS Flow cytometry was used to detect the expression level of ROS and cell apoptosis of 16 HBE cells stimulated by LPS pretreated with 15 and 30μmol/L paeonol.Western blot method was used to detect the expression levels of cytochrome C(Cyt-C),caspase-9,caspase-3,Bax and Bcl-2.RESULTS Compared with the LPS group,the expression level of ROS in 16 HBE cells stimulated by LPS was significantly decreased in both low and high dose paeonol groups(P<0.05),and the early and late cell apoptosis rate and the expression level of Cyt-C in cytoplasm were significantly decreased(P<0.05),while the expression level of Cyt-C in mitochondria was significantly increased(P<0.01).Paeonol significantly decreased the expression of caspase-9 and caspase-3 and the expression of pro-apoptotic protein Bax in 16 HBE cells stimulated by LPS(P<0.01),and significantly increased the expression of anti-apoptotic protein Bcl-2(P<0.05).CONCLUSION Paeonol alleviates the effect of LPS stimulation on airway epithelial cells by inhibiting the mitochondrial apoptosis pathway.