促血小板生成素(TPO)抗化学性缺氧对PC12细胞保护作用的研究-生理学专业论文.docxVIP

ABSTRACT Objectives: Research reorganization boost platelet hormone (thrombopoietin, TPO) in the rat adrenal pheochromocytoma (PC12) cells induced by hypoxia protective effect and possible mechanism. For the clinical treatment of ischemia anoxic brain damage to seek more effective treatment measures and provide some experimental basis. Methods: Using chemical oxygen dose of cobalt chloride (CoCl2) simulation of PC12 cells induced by hypoxia, epoetin boost platelet hormone(thrombopoietin, TPO) establish CoCl2 inducing PC12 cell injury protection model; The application of tetramethyl azo salt colorimetry azole (3 - (4, 5 - dimethyl - 2 - thiazole - 2, 5 - diphenyl tetrazolium bromide, determined by MTT) the cytotoxicity of judgment, the survival rate of PC12 cells were examined; The Annexin Ⅴ/PI double flow cytometry to detect cell apoptosis; The intracellular Reactive Oxygen Species (ROS) : active Oxygen detection Kit (Reactive Oxygen Species Assay Kit), using fluorescent probe DCFH-DA staining fluorescence microscope detection of intracellular Oxygen free radicals (Reactive Oxygen Species, ROS) content; The mitochondrial membrane potential detection: JC-1 staining, the intensity of the red and green fluorescence microscope cell and radiography, using software to calculate the field of vision the ratio of red and green light; Flow cytometry to detect the change of cell membrane potential; The use of SPSS13.0 statistical software to analyze data, data with mean±standard error, said inspection level (P 0.05). Results: CoCl2 inhibit PC12 cell growth has the concentration dependence: chemical oxygen CoCl2 simulation agent can significantly inhibit the growth of PC12 cells, determined by MTT colorimetry to detect, CoCl2 stimulus concentration (200, 400, 600, 800, 1000 ?mol/L) PC12 cells after 24h, respectively, the cell survival rate decreased significantly, therefore, in the subsequent experiments with 500?mol/L for 24h concentration of CoCl2 role as models of chemical