The Relationship Between PRMT5-mediated Methylation And Oxygen-glucose Deprivation-reperfusion Injury

Objective: In this experiment,we established a mouse HT4 cell line subtype(HT22 cell)oxygen deprivation reperfusion model of hippocampal neurons to detect the expression level of PRMT5 protein and GM130 methylation level,and to preliminarily investigate whether there is a correlation between the two changes.Methods: 1.Establishing model: Using HT22 cells as the research object,OGD/R model was established after oxidizing HT22 cells for 6 hour and then cultured under normal culture conditions for 0 hour,6 hour,12 hour,and 24 hour,respectively.2.Experimental grouping: divided into control group,OGD/R group.The OGD/R group was divided into four subgroups(OGD/R0 h,OGD/R6 h,OGD/R12 h,and OGD/R24h)according to the reperfusion time points of 0 hour,6 hour,12 hour,and 24 hour.3.Detection method: Morphological changes of cells were observed under fluorescence microscope using Hoechest33324 staining.The expression of PRMT5 protein was detected by Western Bolt technique.The methylation level of GM130 was detected by co-immunoprecipitation ray counting.Results: 1.Hoechest33324 staining observed by fluorescence microscopy showed that: OGD/R cells prolonged with the reperfusion time,cell apoptosis rate increased,compared with the control group,the difference was statistically significant(P <0.05).2.Western blot analysis showed that the expression of PRMT5 protein in OGD/R cells at 6h,12 h and 24 h after reperfusion was significantly increased(P<0.05).3.The results of co-immunoprecipitation ray counting showed that: OGD/R cells prolonged with the reperfusion time,ray count increased,compared with the control group,the difference was statistically significant(P <0.05).4.Linear regression analysis of apoptosis rate and PRMT5 protein expression in HT22 cells showed that the apoptosis rate of HT22 cells and the expression level of PRMT5 protein may be linearly correlated.5.Linear regression analysis of HT22 cell apoptosis rate and GM130 methylation level showed that there may be a linear correlation between the apoptosis rate of HT22 cells and the GM130 methylation level.6.Linear regression analysis of PRMT5 protein expression and GM130 methylation level showed that there may be linear correlation between PRMT5 protein expression and GM130 methylation level.Conclusion: PRMT5-mediated methylation may be associated with oxygen-glucose deprivation and reperfusion injury in HT22 cells.