帕金森病多巴胺细胞凋亡与Fas基因表达的机制研究

Study on mechanism of dopamine apoptosis and Fas gene expression in Parkinson′s disease

目的探讨Fas基因mRNA抑制后对帕金森病(PD)大鼠模型中多巴胺细胞的抗细胞凋亡作用。方法将75只SD大鼠按随机数字表法分为五组,每组15只。空白对照组:大鼠脑黑质内注入生理盐水;PD组:脑黑质内注射6-OHDA;Fas siRNA组:脑黑质内注射Fas基因抑制剂Fas siRNA,2 d后注射同剂量的6-OHDA;阳性对照组及阴性对照组:脑黑质内分别注射Fas siRNA阳性对照物、阴性对照物,2 d后注射同剂量的6-OHDA。采用免疫组化、聚合酶链反应和蛋白印迹杂交技术检测多巴胺阳性细胞数、Fas-mRNA及Fas-蛋白相对表达量。结果PD组大鼠的TH阳性细胞计数少于空白对照组(P<0.05);Fas siRNA组和阳性对照组大鼠的TH阳性细胞计数多于PD组(P<0.05);大鼠的TH阳性细胞计数多于阴性对照组(P<0.05)。PD组大鼠的Fas-mRNA及Fas-蛋白相对表达量均高于空白对照组(P<0.05);Fas siRNA组和阳性对照组大鼠的Fas-mRNA及Fas-蛋白相对表达量均低于PD组(P<0.05);Fas siRNA组和阳性对照组大鼠的Fas-mRNA及Fas-蛋白相对表达量均低于阴性对照组(P<0.05)。结论Fas基因的表达被抑制后可有效减少多巴胺细胞的凋亡。

Objective To explore the antiapoptosis effect on dopamine cell in the rat model of Parkinson′s disease(PD)after Fas gene mRNA inhibition.Methods Seventy-five SD rats were divided into 5 groups by random number table method,with 15 rats in each group.Blank control group:normal saline was injected into the substantia nigra of rats.PD group:intracerebral injection of 6-OHDA.In Fas siRNA group:Fas gene inhibitor Fas siRNA was injected into the substantia nigra of brain,and 6-OHDA of the same amount was injected 2 days later.Positive control group and negative control group:Fas siRNA positive control and negative control were injected into the substantia nigra respectively,and 6-OHDA of the same measurement was injected 2 days later.Immunohistochemistry,PCR and Western blot were used to detect the number of dopamine positive cells,Fas mRNA and the relative expression of Fas protein.Results The number of TH positive cells in PD group was less than that in blank control group(P<0.05).The number of TH positive cells in FAS siRNA group and positive control group was higher than that in PD group(P<0.05).The number of TH positive cells in FAS siRNA group and positive control group was more than that in negative control group(P<0.05).The relative expression of Fas-mRNA and Fas-protein in PD group was higher than that in blank control group(P<0.05).The relative expression of Fas-mRNA and Fas-protein in FAS siRNA group and positive control group were lower than that in PD group(P<0.05).The relative expression of Fas-mRNA and Fas-protein in Fas siRNA group and positive control group was lower than that in negative control group(P<0.05).Conclusion The dopamine apoptosis can be reduced effectively after the inhibition of expression of Fas gene.