人参皂苷Rg1对帕金森病模型小鼠黑质区FADD和FLIP表达的影响及其意义

摘要

Objective To investegate the effect of ginsenoside Rg1 on the apoptosis related protein FLICE-inhibitory protein(FLIP),Fas-associated death domain protein (FADD)and Caspase-3 in the subatania nigra(SN)of 1-methyl-4-phenyl-1,2,3,6-tetrahyd-ropyridine (MPTP)-induced mouse models of Parkinson’s disease(PD), and to investigate the role of FADD and FLIP in the pathogenesis of PD and the protective effect of ginsenosides Rg1 on dopaminergic neurons.Methods 45 C57BL/6N mice were randomly divided into control group,model group and ginsenoside Rg1 group (n=15).The mice in model group were injected with MPTP by intraperitoneal,the mice in Rg1 group were injected with ginsenoside Rg1 before injecting MPTP,and the mice in control group were injected with normal saline by intraperitoneal. The behavioral changes of the mice in various groups were observed, and immunohistochemistry and Western blotting methods were used to observe the expressions of tyrosine hydroxylase (TH),FADD,FLIP and Caspase-3 in substantia nigra of the mice.Results Compared with control group,the mice in model group presented with typical symptoms of PD, the TH-positive neurons in the subatania nigra was significantly reduced (P<0.01 ), the number of FADD, FLIP and Caspase-3 positive cells was significantly increased(P<0.01),and the cytoplasm was deeply stained;the protein expression levels of FADD,FLIP and Caspase-3 were significantly increased (P<0.01).Compared with model group,the PD symptoms of the mice in ginsenoside Rg1 group reduced, the number of TH-positive neurons was significantly increased, the number of positive cells of FLIP,FADD and Caspase-3 were significantly reduced(P<0.01),and the cytoplasm was lightly stained;the protein expression levels of FADD, FLIP and Caspase-3 were significantly reduced (P<0.01 ). Nonlinear correlation analysis found that there was a positive relationship between the number of FADD and Caspase-3 positive cells (r=0.791,P<0.05).Conclusion Ginsenoside Rg1 may play a neural protective effect dopaminergic on neurons by modulating the FADD and FLIP expressions in SN of PD model mice.%目的：探讨人参皂苷 Rg1对帕金森病（PD）模型小鼠黑质区凋亡信号蛋白 Fas 死亡结构域蛋白（FADD）、FADD样白细胞介素1-转化酶样抑制蛋白（FLIP）及Caspase-3表达的影响，阐明FADD和FLIP在PD发病机制中的作用及人参皂苷 Rg1对多巴胺（DA）能神经元的保护作用。方法：45只C57BL/6N小鼠随机分为对照组、模型组和人参皂苷 Rg1组，每组15只。模型组小鼠腹腔注射1-甲基-4-苯基-1，2，3，6-四氢吡啶（MPTP），人参皂苷 Rg1组小鼠注射 MPTP前先注射人参皂苷 Rg1，对照组小鼠腹腔注射生理盐水。观察各组小鼠行为学变化；采用免疫组织化学法和免疫蛋白印迹法检测各组小鼠中脑黑质区酪氨酸羟化酶（TH）、FLIP、FADD及Caspase-3的表达。结果：与对照组比较，模型组小鼠出现典型 PD症状，黑质区 TH 阳性细胞数明显减少（P＜0.01），FADD、FLIP 及 Caspase-3阳性细胞数明显增加（P＜0.01），胞浆染色深；FADD、FLIP 及Caspase-3蛋白表达水平升高（P＜0.01）。与模型组比较，人参皂苷 Rg1组小鼠 PD症状减轻，黑质区 TH 阳性细胞数明显增多（P＜0.01），FADD、FLIP及Caspase-3阳性细胞数明显减少（P＜0.01），胞浆染色浅；FADD、FLIP和Caspase-3蛋白表达水平降低（P＜0.01）。FADD 与 Caspase-3阳性细胞数呈正相关关系（r＝0.791， P＜0.05）。结论：人参皂苷 Rg1可通过影响PD模型小鼠黑质区 FADD和 FLIP的表达对 DA能神经元起到一定的保护作用。