MiRNA-29b Regulation Of COL4A1 On The Protective Mechanism Of Ischemic Stroke Nerve Injury

Objective: The protective effect of mir-29 b on the hypoxia/ reoxygenation injury N2 a cells was desmonstrated by constructing on the oxygen-sugar deprivation/reoxygenation model(OGD/R)of the mouse-derived neuroblastoma(N2a cells),and the potential protection mechanism was discussed.The type IV collagen a1(COL4A1)was verified as the target of the Mir-29 b.through the establishent of the OGD/R model of the N2 a cell,the mir-29 b is proved to be protective effect on the OGD/R injury of the N2 a cell by targeting the expression of the COL4A1.Methods: N2 a cells were cultured in normal cell culture environment and oxygen-glucose deprivation/reoxygenation environment,and the expression of mir-29 b in N2 a cells cultured in two different environments was detected by q PCR.The cultured N2 a cells were divided into three groups according to different treatments.The cells were divided into three groups according to different treatments: no intervention group(NC),was given mir-29 b mimics intervention negative control group(mir-29 b mimics NC),mi RNA-29 b mimimics intervention group(NC).CCK8 and Cell clone formation was used to detect the proliferation of cells,Hoechst and flow cytometry was used to detect the apoptosis of cells,and Western Blot was used to detect the expression of apoptosis-related proteins in the two groups.The expression of COL4A1 in three groups of cells treated with different treatments was detected by q PCR,and the wild and mutant vectors of COL4A1 were constructed,and the target genes were verified by double luciferase.The synthetic COL4A1 vector was designed and synthesize.The synthesized COL4A1 was transferred into N2 a cells and divided into(NC),COL4A1 negative control group(COL4A1 NC)and COL4A1 group(COL4A1);CCK8 and clone formation detected the proliferation of the above three groups of cells,Hoechst and flow cytometry detected the apoptosis of the above three groups of cells,Western Blot to detect the expression of apoptosis-related proteins in the above three groups.Results:The expression of mir-29 b in N2 a cells cultured with hypoxia/reoxidation was lower than that in N2 a cells cultured with normal culture.After upregulating mir-29 b,the proliferation ability of N2 a cells was significantly enhanced and the apoptosis of N2 a cells was significantly inhibited.After upregulation of mir-29 b,the expression of COL4A1 was inhibited,while the proliferation of COL4A1 control group was significantly higher than that of overexpression group,and the apoptosis of COL4A1 control group was lower than that of overexpression group.Conclusion:Mir-29 b can regulate the damage of ischemia/hypoxia reperfusion to nerve cells by reducing the expression of COL4A1.