论文部分内容阅读
目的建立氯化钴(CoCl2)诱导大鼠原代脑微血管内皮细胞(BMECs)缺氧模型,并观察细胞缺氧损伤情况。方法使用不同浓度(100、200、400、800μmol/L)CoCl2诱导大鼠原代BMECs缺氧,蛋白印迹法检测低氧诱导因子-1α(HIF-1α)蛋白水平来反映细胞缺氧的程度,同时采用CCK-8法、化学比色法及TUNEL法检测细胞增殖、氧化应激和细胞凋亡水平。结果 200μmol/L CoCl2处理大鼠原代BMECs 6h后,HIF-1α蛋白水平较常氧阴性对照组显著升高(P<0.01);随着CoCl2处理浓度(200、400、800μmol/L)的增加,与常氧阴性对照组相比,细胞增殖抑制率逐渐升高,MDA水平明显增高(P<0.01),GSH水平和SOD活性显著降低(P<0.01),细胞凋亡率明显增加(P<0.01)。结论 200μmol/L浓度CoCl2处理大鼠原代BMECs 6h可以诱导明显的低氧HIF反应,而细胞增殖、氧化应激和细胞凋亡的损伤较轻,是模拟大鼠原代BMECs细胞缺氧的理想条件。
Objective To establish a rat model of primary cerebral microvascular endothelial cells (BMECs) -induced hypoxia induced by cobalt chloride (CoCl2), and to observe the cell injury induced by hypoxia. Methods Hypoxia was induced in primary BMECs of rats by CoCl2 at different concentrations (100, 200, 400 and 800μmol / L), and the level of HIF-1α protein was detected by Western blotting to reflect the degree of cell hypoxia. At the same time, CCK-8 method, chemical colorimetry and TUNEL method were used to detect cell proliferation, oxidative stress and apoptosis. Results After treated with 200μmol / L CoCl 2 for 6h, the level of HIF-1α protein in BMECs was significantly increased compared with normoxia-negative control group (P <0.01). With the increase of CoCl2 concentration (200,400,800μmol / L) (P <0.01). GSH level and SOD activity were significantly decreased (P <0.01), and the apoptosis rate was significantly increased (P <0.01). Compared with normoxia-negative control group, the cell proliferation inhibition rate increased gradually, MDA level increased significantly 0.01). Conclusions Treatment of primary BMECs with 200μmol / L of CoCl2 for 6h can induce obvious hypoxic HIF response, but less damage of cell proliferation, oxidative stress and apoptosis, which is ideal for hypoxia of primary BMECs in rats condition.