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目的:探讨大气细颗粒物(PM2.5)对EA.hy926型人脐静脉内皮细胞的损伤及黄连素的保护作用及其机制。方法:采集大气PM2.5并分别以0、20、200、400 mg/L染毒EA.hy926细胞24 h,MTT法检测细胞存活率、流式细胞术检测细胞凋亡、Western blot法检测p-ERK1/2、BAX、BCL-2蛋白表达,ELISA法测IL-6、TNF-α、MDA含量及SOD、LDH活性;分别加入黄连素(10、50、100μmol/L)和ERK1/2通路特异性阻滞剂PD98059 20μmol/L检测黄连素的干预作用。结果:与对照组比较,PM2.5染毒后呈剂量依赖性降低细胞存活率,并上调p-ERK1/2蛋白水平及BAX/BCL-2蛋白比率以促进细胞凋亡、诱导分泌IL-6、TNF-α及升高MDA含量、降低SOD活性、升高LDH活性(P<0.05);黄连素呈剂量依赖性升高PM2.5作用下细胞存活率、下调p-ERK1/2蛋白水平及BAX/BCL-2蛋白比率以抑制细胞凋亡、降低IL-6、TNF-α及MDA含量、升高SOD活性、降低LDH活性(P<0.05)。结论:黄连素能通过抑制ERK1/2通路,减轻PM2.5对EA.hy926细胞的损伤。
Objective: To investigate the effects of PM2.5 on the injury of EA.hy926 human umbilical vein endothelial cells and the protective effect of berberine and its mechanism. Methods: PM2.5 was collected and the cells were treated with 0, 20, 200 and 400 mg / L for 24 h respectively. The cell viability was detected by MTT assay and the apoptosis was detected by flow cytometry. The expression of p The levels of IL-6, TNF-α, MDA and the activities of SOD and LDH were measured by ELISA. The expressions of ERK1 / 2 Specific Inhibitor PD98059 20μmol / L Intervention of Berberine. Results: Compared with the control group, PM2.5 dose-dependently decreased the cell viability and up-regulated the protein level of p-ERK1 / 2 and the ratio of BAX / BCL-2 protein in order to promote cell apoptosis and induce the secretion of IL-6 (P <0.05); Berberine could increase the cell survival rate and down-regulate the protein level of p-ERK1 / 2 in a dose-dependent manner under the effect of PM2.5 and increase the content of MDA and decrease the activity of SOD and the activity of LDH BAX / BCL-2 protein ratio could inhibit the apoptosis, decrease the levels of IL-6, TNF-α and MDA, increase the activity of SOD and decrease the activity of LDH (P <0.05). Conclusion: Berberine can reduce the injury of EA.hy926 cells by inhibiting the ERK1 / 2 pathway.