【摘要】 目的 建立犬肾细胞（Madin-Darby canine kidney, MDCK）缺氧模型，进一步探讨丝裂原活化蛋白激酶（MAPKs）在缺氧性细胞损伤中的活性改变。方法 将MDCK细胞置于体积分数为5%CO₂和95%N₂的有机玻璃调节性密闭容器中分别培养24、36、48、72、84 h, MTT实验检测细胞的存活力，蛋白免疫印迹法（Western blot）检测JNK（c-Jun NH2 terminal protein kinase）和p38 MAPK的磷酸化活性。结果 MDCK细胞缺氧24、36、48、72、84 h后，与正常对照组相比，MTT D值均明显下降（P<0.01）。MDCK细胞缺氧后，JNK和p38 MAPK的磷酸化水平增高。结论 此方法可以成功建立操作简便、有效的MDCK细胞缺氧模型，且是通过激活JNK、p38 MAPK引起缺氧性细胞损伤。更多还原

【Abstract】 Objective To establish a hypoxic model of Madin-Darby canine kidney（MDCK） cells and to investigate the changes of mitogen activated protein kinase（MAPKS） activity during hypoxic cell injury. Methods MDCK cells were incubated in a modular incubator chamber（5% CO₂ and 95% N₂） for 24 h, 36 h, 48 h, 72 h and 84 h. Cell viability was measured by MTT assay. The phosphorylation of JNK and p38 MAPK was detected by Western blotting. Results Compared with the control, MDCK cells showed significantly decreases OD value in MTT assay after cultivated in hypoxic condition for 24 h, 36 h, 48 h, 72 h and 84 h（P<0.01）. Meanshile, the phosphorylation of JNK and p38MAPK was enhanced under hypoxic condition. Conclusion A simple and effective model of hypoxic MDCK cells was established by the present method, where hypoxia cell damages can be induced by activation of JNK and p38MAPK.更多还原