小檗碱对PM2.5诱导的血管内皮细胞损伤的抑制作用及其p38 MAPK通路...

来自 : qikan.cqvip.com/Qikan/Article/ 发布时间：2021-03-25

目的：研究小檗碱对大气中空气动力学直径≤2.5μm的细颗粒物（PM2.5）诱导的EA.hy926型人脐静脉内皮细胞（简称EA.hy926细胞）损伤的抑制作用,以及其p38分裂原激活蛋白激酶（p38 MAPK）信号通路机制。方法：采集大气PM2.5并以0（空白对照）、20、200、400 mg/L孵育EA.hy926细胞24 h,测定细胞存活率、凋亡率、细胞中白细胞介素6（IL-6）、肿瘤坏死因子α（TNF-α）、丙二醛（MDA）含量,超氧化物歧化酶（SOD）、乳酸脱氢酶（LDH）活性,以及磷酸化p38 MAPK（p-p38 MAPK）、B淋巴细胞癌2（Bcl-2）、Bcl-2相关X蛋白（Bax）的蛋白水平。同法测定空白对照组、PM2.5组（200 mg/L PM2.5）、p38 MAPK通路特异性阻滞剂SB203580组（20μmol/L SB203580＋200 mg/L PM2.5）和小檗碱低、中、高浓度组（5、10、20μmol/L小檗碱＋200 mg/L PM2.5）EA.hy926细胞的上述指标。结果：与空白对照比较,200、400 mg/L PM2.5孵育细胞后细胞存活率、SOD活性、Bcl-2蛋白表达均明显降低;细胞凋亡率,IL-6、TNF-α、MDA含量,LDH活性以及p-p38 MAPK、Bax蛋白水平均明显升高（P〈0.05）,效果呈浓度依赖性。与PM2.5组比较,小檗碱中、高浓度组和SB203580组细胞存活率、SOD活性、Bcl-2蛋白表达明显升高;细胞凋亡率,IL-6、TNF-α、MDA含量,LDH活性以及p-p38 MAPK、Bax蛋白水平均明显降低（P〈0.05）。结论：小檗碱可通过抑制p38 MAPK通路,减轻PM2.5对EA.hy926细胞的损伤。 OBJECTIVE： To study the inhibitory effects of berberine on EA.hy926 human umbilical vein endothelial cells （EA. hy926 cells） injury induced by particulates with no more than 2.5 μm air aerodynamic diameter in atmospheric （PM2.5）, and its p38 mitogen-activated protein kinase （MAPK） signal pathway mechanism. METHODS： PM2.5 samples were collected and hatched EA.hy926 cells with concentrations of 0 （blank control）, 20, 200 and 400 mg/L for 24 h. The survival rate and apoptosis rate of cells, contents of IL-6, TNF-ct and MDA, activities of SOD and LDH, protein levels of p-p38 MAPK, Bcl-2 and Bcl-2 associated X protein （Bax） were detected. The above indexes of EA.hy926 cells in blank control group, PM2.5 group （200 mg/L PM2.5）, p38 MAPK pathway-specific blocker SB203580 group （20 μmol/L SB203580＋200 mg/L PM2.5）, berberine low-, medium- and high-concentrations groups （5, 10,20μmol/L berberine＋200 mg/L PM2.5） were also determined. RESULTS： Compared with blank control, survival rate of cells, SOD activity and Bcl protein decreased after 200, 400 mg/L PM2.5 hatched; apoptosis rate of cells, contents of IL-6, TNF-α and MDA, LDH activity, protein levels of p-p38 MAPK and Bax increased （P〈0.05）, in concentra- tion-dependent manner. Compared with PM2.5 group, survival rate of cells, SOD activity and Bcl-2 protein increased in berberine medium-, high-concentrations groups and SB203580 group; apoptosis rate of cells, contents of IL-6, TNF-α and MDA, LDH ac- tivity, protein levels of p-p38 MAPK and Bax decreased （P〈0.05）. CONCLUSIONS： Berberine attenuates PM2.5-induced EA. hy926 cells injury via the inhibition of p38 MAPK pathway.