Effect of over-expression of BIRC5 gene on the activity and VEGF expression of cerebral microvascular endothelial cells induced by oxygen glucose deprivation/reoxygenation
Objective To investigate the protective effect of over-expression of BIRC5 gene on microvas-cular endothelial cell injury induced by oxygen glucose deprivation/reoxygenation(OGD/R)in mouse brain,and to analyze its possible mechanism.Methods According to different intervention methods,mouse brain microvascular endothelial cells were divided into normal control group(normal cell culture),cell damage group(normal cell culture for 24 hours,followed by OGD3h/R3h injury),BIRC5 intervention group(The cells were transfected with BIRC5 plasmids of adenovirus and cultured for 24 hours and then underwent OGD for 3 hours/R for 3 hours.)and negative control group(The cells were transfected with empty plasmids of adenovirus and cultured for 24 hours and then underwent OGD for 3 hours/R for 3 hours.).Laser confocal microscope was used to observe the morphological changes of cells in each group.The cell survival rate and apoptosis rate were detected by MTT and flow cytometry,respectively.The mRNA and protein expressions of BIRC5 and VEGF were detected by RT-PCR and Western blot,respectively.Results The cytoskeletal microfilaments of the normal control group were connected with each other and regularly distributed in an orderly network.In the in-jury group and negative control group,the microfilaments of the cells were broken,and the contraction of the cells became shorter or moved to the periphery;the microfilaments were arranged in a disordered network;the cell appearance was shrunk,the gap enlarged and a small number of microfilaments missing with gaps;In the BIRC5 intervention group,the microfilaments of cells were connected,spindle shaped with regular arrange-ment and reduced gap between cells,indicating that the overexpression of BIRC5 gene could reduce the disor-der of cytoskeletal microfilaments of damaged cells.Compared with the normal control group,the cell damage group,the BIRC5 intervention group and the negative control group had significantly reduced cell survival rates,but had significantly increased apoptosis rates,showing significant differences(P＜0.05).Compared with those of the cell damage group,the cell survival rate of the BIRC5 intervention group was significantly in-creased,while the apoptosis rate was significantly reduced,and the differences were statistically significant(P＜0.05).As compared with normal control group,BIRC5 and VEGF mRNA and protein in cell damage group,BIRC5 intervention group and negative control group was significantly reduced,and the difference was statisti-cally significant(P＜0.05).Compared with the cell damage group,the BIRC5 intervention group had signifi-cantly increased mRNA and protein expressions of BIRC5 and VEGF,showing siginifcant differences(P＜0.05).Conclusion Overexpression of the BIRC5 gene has a protective effect on microvascular endothelial cell injury induced by oxygen glucose deprivation/reoxygenation,and the mechanism may be related to its up-regulation of VEGF expressions,suggesting that BIRC5's regulation of VEGF expressions to promote angio-genesis may be one of the important mechanisms for the establishment and formation of good cerebral collateral circulation.
overexpression of BIRC5 genecerebral microvascular endothelial cellssurvivalapoptosisoxygen and sugar deprivation/reoxygenationvascular endothelial growth factor
万方数据
维普
