c-Jun和Bcl-2在海洛因所致大鼠小脑颗粒细胞凋亡中的作用

Effects of c-Jun and Bcl-2 in diacetylmorphine-induced apoptosis in rat cerebellar granule cells

目的探究海洛因致小脑颗粒细胞凋亡中c-Jun氨基端激酶(c-Jun)和B淋巴细胞瘤-2基因(Bcl-2)蛋白质的表达情况,分析c-Jun和Bcl-2与海洛因影响小脑颗粒细胞凋亡的关系。方法取8只出生7d的SD大鼠小脑颗粒细胞进行体外培养,随机分为实验组(以80 mg/L海洛因干预)、抑制剂组(以80 mg/L海洛因和10μmol/L CEP1347干预)、正常组(正常培养的小脑颗粒细胞)。采用Hoechst 33258荧光染色技术检测细胞凋亡率,采用免疫荧光、Western blot检测c-Jun和Bcl-2蛋白的表达情况。结果实验组细胞凋亡率较抑制剂组及正常组高(P<0.05);实验组与正常组比较,c-Jun蛋白含量明显升高(P<0.05),Bcl-2蛋白含量下调(P<0.05);抑制剂组中c-Jun蛋白含量较实验组明显下降(P<0.05)。结论 c-Jun和Bcl-2参与海洛因致小脑颗粒神经元细胞凋亡过程,并且可能是c-Jun氨基末端激酶信号通路中的重要候选促凋亡因子。

Objective To investigate the expression of c-Jun amino-terminal kinase(c-Jun)and B-cell lymphoma-2 gene(Bcl-2)proteins in diacetylmorphine-induced apoptosis in cerebellar granule cells and to analyze the relationship between c-Jun and Bcl-2 and diacetylmorphine-induced apoptosis.Methods The cerebellar granulosa cells from SD rats were cultured in vitro.Divide them into 80 mg/L diacetylmorphine intervention group,80 mg/L diacetylmorphine and 10μmol/L CEP1347 intervention group and control group randomly.Detect the apoptosis rate by Hoechst 33258 technique and the expression of c-Jun and Bcl-2 protein by immunofluorescence and western blotting.Results The apoptosis rate of 80 mg/L diacetylmorphine intervention group was higher than that of the control group(P <0.05).The content of c-Jun protein in the 80 mg/L diacetylmorphine intervention group was significantly higher than that in the control group(P <0.05),and the content of Bcl-2 protein was down-regulated(P <0.05).The contents of cJun protein in 80 mg/L diacetylmorphine and in 10μmol/L CEP1347 intervention group were significantly lower than that in 80 mg/L diacetylmorphine intervention group(P <0.05).Discussion The results showed that c-Jun and Bcl-2 were involved in diacetylmorphine-induced apoptosis of cerebellar granule cells and may be the important candidate apoptosis factors in c-Jun amino-terminal kinase signaling pathway.