Effects of calycosin on mitochondrial apoptotic pathway in oxygen-glucose deprivation / reoxygenation PC12 cells / 解剖学报

ABSTRACT

Objective To investigate the effect of calycosin on mitochondrial apoptotic pathway in oxygen-glucose deprivation/ reoxygenation PC12 cells. Methods PC12 cells were randomly divided into four groups control group, model group, calycosin group and nimodipine group. Except for the control group, the other groups were treated with oxygen and glucose deprivation for 2 hours and compound oxygen and glucose for 24 hours. Calycosin group and nimodipine group were treated with drug-containing medium containing calycosin (0. 07 μmol/ L) and nimodipine (5. 00 μmol/ L) simultaneously with reoxygenation. CCK-8 method was used to detect cell survival rate, flow cytometry was used to detect cell apoptosis rate, immunofluorescence method was used to detect Bax/ Bcl-2 ratio, Western blotting was used to detect the expression of key proteins cytochrome C (Cyt-C), apoptotic protease activating factor-1 (Apaf-1) and Caspase-3 in mitochondrial apoptotic pathway. Results Compared with the control group, the survival rate of cells in model group decreased significantly (P<0. 05), and the apoptotic rate increased significantly (P<0. 05), the ratio of Bax/ Bcl-2 was significantly increased (P<0. 05), and the expression of key proteins Cyt-C, Apaf-1 and Caspase-3 in mitochondrial apoptotic pathway were significantly increased (P<0. 05). Compared with the model group, the cell survival rates of calycosin group and nimodipine group increased significantly (P < 0. 05), apoptotic rate decreased significantly (P<0. 05), the ratio of Bax/ Bcl-2 was significantly decreased (P<0. 05), and the expression of key proteins of mitochondrial apoptotic pathway, Cyt-C, Apaf-1 and Caspase-3 were significantly decreased (P < 0. 05). The difference has statistical significance. Conclusion Calycosin can significantly improve the survival rate of oxygen-glucose deprivation/ reoxygenation PC12 cells and inhibit cell apoptosis. Its mechanism is closely related to the inhibition of the expression of key proteins Cyt-C, Apaf-1 and Caspase-3 in mitochondrial apoptotic pathway by calycosin.