miRNA-132靶向SCN2A调控对癫痫大鼠海马区细胞凋亡的影响

Effects of microRNA-132 targeting SCN2A regulation on apoptosis in hippocampus of epileptic rats

目的研究miRNA-132靶向SCN2A调控对癫痫大鼠海马区细胞凋亡的影响。方法清洁级健康雄性SD大鼠60只,随机分为模型组、过表达组、沉默组各10只。细胞转染,建立沉默组、过表达组。荧光定量检测miRNA-132、SCN2A表达,MTT检测细胞增殖能力,采用流式细胞仪检测细胞凋亡,Western blot检测Bcl-2、Bax、Akt表达。结果沉默组SCN2A mRNA表达高于模型组和过表达组(P<0.05)。沉默组海马区细胞增殖率高于模型组和过表达组,细胞凋亡率低于模型组和过表达组(P<0.05)。沉默组在24,48,72 h细胞增殖率逐渐升高,过表达组相反(P<0.05)。沉默组在24,48,72 h细胞凋亡率逐渐降低,过表达组相反(P<0.05)。沉默组Bcl-2蛋白表达低于模型组和过表达组,Bax、Akt表达高于模型组和过表达组(P<0.05)。结论miRNA-132沉默能够增强SCN2A表达,miRNA-132通过靶向调控SCN2A,能促进癫痫大鼠海马区细胞增殖,抑制其凋亡。

Objective To study the effect of microRNA-132 targeting SCN2A on apoptosis in hippocampus of epileptic rats.Methods Total of 60 healthy male SD rats of clean grade were randomly divided into model group,over-expression group and silence group,with 10 rats in each group.Cellular transfection,silence group and over-expression group were also established.The expression of miRNA-132 and SCN2A was quantitatively detected by fluorescence,the proliferation of cells was detected by MTT,the apoptosis was detected by flow cytometry,and the expression of Bcl-2,Bax and Akt was detected by Western blot(WB).Results The expression of SCN2A in silence group was higher than that in model group and over-expression group(P<0.05).The proliferation rate of hippocampal cells in silence group was higher than that in model group and over-expression group,and the apoptosis rate was lower than that in model group and over-expression group(P<0.05).The proliferation rate of cells in silence group increased gradually at 24,48 and 72 hours,while that in over-expression group was the opposite(P<0.05).The apoptotic rate of the silent group decreased gradually at 24,48 and 72 hours,while that of the over-expressed group decreased gradually(P<0.05).The expression of Bcl-2 protein in silence group was lower than that in model group and over-expression group,and the expression of Bax and Akt was higher than that in model group and over-expression group(P<0.05).Conclusion The silencing of microRNA-132 can enhance the expression of SCN2A.MicroRNA-132 can promote the proliferation and inhibit the apoptosis of hippocampal cells in epileptic rats by targeting SCN2A.