丹参酮IIA通过抑制p38 MAPK通路减轻PM2.5对血管内皮细胞的损伤

 目的: 探讨PM2.5对EA.hy926型人脐静脉内皮细胞损伤的影响及丹参酮ⅡA在这一过程中的作用及机制。方法: 采集广州城区大气PM2.5并以不同质量浓度(0、20、200、400 mg/L)染毒EA.hy926细胞24 h,MTT法测细胞存活率,流式细胞术测细胞凋亡,Western blot法测p-p38 MAPK、Bax和Bcl-2的蛋白水平,ELISA法测白细胞介素-6(IL-6)及肿瘤坏死因子-α(TNF-α)含量,并测定细胞丙二醛(MDA)含量、超氧化物歧化酶(SOD)及乳酸脱氢酶(LDH)活性;分别加入丹参酮ⅡA(5、10、20μ mol/L)和p38 MAPK通路特异性阻滞剂SB20358020μ mol/L检测丹参酮ⅡA的干预作用及机制。结果: 与对照组比较,PM2.5染毒后呈剂量依赖性降低EA.hy926细胞的存活率,上调p-p38 MAPK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及TNF-α,降低SOD活性,增加MDA含量及LDH活性,差异均有统计学显著性(P<0.05);丹参酮ⅡA呈剂量依赖性增加EA.hy926细胞的存活率,下调p-p38 MAPK蛋白水平及Bax/Bcl-2蛋白比率以抑制细胞凋亡,降低IL-6及TNF-α含量,增加SOD活性,降低MDA含量及LDH活性,差异均有统计学显著性(P<0.05)。结论: 丹参酮ⅡA可通过抑制p38 MAPK通路,减轻PM2.5对EA.hy926细胞的损伤。

Tanshinone ⅡA attenuates PM2.5-induced vascular endothelial cell injury via p38 MAPK signal pathway

AIM: To investigate the effect and the mechanism of tanshinone ⅡA in attenuating PM2.5-induced human umbilical vein endothelial EA.hy926 cell injury. METHODS: The samples of fine particulate matter(PM2.5) were collected in Guangzhou and made into suspension. Different concentrations(0, 20, 200 and 400 mg/L) of PM2.5 were added to EA.hy926 cells. The viability and apoptosis of EA.hy926 cells, the protein levels of p-p38 MAPK, Bax and Bcl-2 in the EA.hy926 cells, the contents of interleukin-6(IL-6), tumor necrosis factor-α(TNF-α) and malonaldehyde(MDA), and the activity of superoxide dismutase(SOD) and lactic dehydrogenase(LDH) in the EA.hy926 cell culture supernatant were measured by MTT assay, flow cytometry, Western blot, ELISA and colorimetry, respectively. Tanshinone ⅡA at different concentrations(5, 10 and 20μmol/L) or a specific inhibitor of p38 MAPK pathway, SB203580(20μmol/L), was added into the EA.hy926 cells to observe the effect of tanshinone ⅡA. RESULTS: Compared with control group, PM2.5 significantly increased the apoptosis, the contents of IL-6, TNF-α and MDA, the activity of LDH, and the protein levels of p-p38 MAPK and Bax/Bcl-2 ratio, but decreased the viability and SOD activity in the EA.hy926 cells(P<0.05). Compared with PM2.5 group, tanshinone ⅡA significantly decreased the apoptosis, the contents of IL-6, TNF-α and MDA, the activity of LDH, and the protein levels of p-38 MAPK and Bax/Bcl-2 ratio, but increased the viability and SOD activity in the EA.hy926 cells(P<0.05). CONCLUSION: Tanshinone ⅡA attenuates PM2.5-induced EA.hy926 cell injury via the inhibition of p38 MAPK pathway.