Effects Of CDK1 On GRASP65 And Golgi Morphology In Oxygen-glucose Deprivation Followed By Reperfusion

Object:To elucidate the effects of Cdk1 on GRASP65 and Golgi Morphology in Oxygen-glucose deprivation reperfusion.Method:1.Employed a stable OGD/R model to simulate ischemic-like condition on PC12 cells.2.Experimental groups were divided into control group,OGD/R group and OGD/R+Roscovitine group.3.The cellular viability and the rate of cell apoptosis were measured by CCK-8 kit and flow cytometric analysis.4.Using immunofluorescence to observe the morphological chang of Golgi apparatus.5.Using western blot to detect the expression of CDK1,GRASP65 and pGRASP65.Results:1.The CCK-8 results showed that cellular viability of OGD/R group were reduced at 6-hour,12-hour and 24-hour time points recovery from oxygen-glucose deprivation,compared with control group at the same time point(P < 0.05);Roscovitine obviously promoted cell activity at 12-hour compared with that in OGD/R group(P<0.05).2.Flow cytometric analysis result showed that cell apoptosis of OGD/R group were increased at 6-hour,12-hour and 24-hour time points recovery from oxygen-glucose deprivation,compared with control group at the same time point(P < 0.05);Roscovitine obviously reduced cell apoptosis compared with that in OGD/R group at the same time point(P<0.05).3.The expression of CDK1 were obviously increased at 6-hour,12-hour and 24-hour time points after OGD/R,compared with the control group(P<0.05);After the intervention of Roscovitine,expression of CDK1 obviously decreased,compared with OGD/R group(P <0.05).The expression of GRASP65 showed no significant differences in all of the groups.The levels of pGRASP65 were obviously increased at 6-hour,12-hour and 24-hour time points after OGD/R,compared with the control group(P<0.05);After the intervention of Roscovitine,expression of pGRASP65 obviously decreased,compared with OGD/R group(P<0.05).4.The immunofluorescence results showed that Golgi apparatus showed no obvious fragmentation at 0-hour and 6-hour time points after OGD/R,while it fragmented seriously at 12-hour and 24-hour time points after OGD/R;After the intervention of Roscovitine,the degree of Golgi fragmentation alleviated.Conclusion:CDK1 mediated GRASP65 phosphorylation after oxygen glucose deprivation reperfusion may be one of the mechanisms of Golgi fragmentation.