芍药甘草汤加减对MPTP诱导的帕金森病模型小鼠的影响

Effect of Modified Shaoyao Gancao Tang on MPTP-induced Parkinson's Disease Model Mice

目的:研究芍药甘草汤加减对1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)诱导的帕金森病模型小鼠的作用。方法:将60只C57/BL6小鼠随机分为正常组,模型组,美多巴组(50 mg·kg^(-1))和芍药甘草汤加减低、中、高剂量组(1,2,4 g·kg^(-1))。将C57/BL6小鼠腹腔注射MPTP(40 mg·kg^(-1)),每天1次,连续7 d。除正常组和模型组每天灌胃生理盐水外,给药组每天灌胃1次相应的药物,灌药第3天后按上述方法造模(除正常组外),给药15 d后,检测各组小鼠的行为学(爬杆实验、游泳实验和悬尾实验)。采用酶联免疫吸附测定(ELISA)检测各组小鼠血清中胱抑素-C(Cys-C)和神经元特异烯醇化酶(NSE)的含量,采用分光光度法检测各组小鼠纹状体中超氧化物歧化酶(SOD),丙二醛(MDA)和谷胱甘肽过氧化物酶(GSH-PX)的水平,采用蛋白免疫印迹法(Western blot)检测各组小鼠纹状体中α-突触核蛋白(α-syn),酪氨酸羟化酶(TH),B淋巴细胞瘤-2基因(Bcl-2)和Bcl-2相关X蛋白(Bax)的表达。结果:与正常组比较,模型组小鼠的行为学实验中游泳实验得分和悬挂实验得分显著降低(P <0. 01),爬杆时间显著增加(P <0. 01),血清中Cys-C含量显著减少(P <0. 01),NSE含量明显增加(P <0. 05),纹状体中SOD水平和GSH-Px水平显著减少(P <0. 01),MDA水平明显增加(P <0. 05),TH表达和Bcl-2表达显著减少(P <0. 01),α-syn和Bax表达明显增加(P <0. 05);与模型组比较,芍药甘草汤加减各剂量组与美多巴组的行为学实验中游泳实验得分和悬挂实验得分均明显增加(P <0. 05),爬杆时间明显减少(P <0. 05),血清中Cys-C含量显著增加(P <0. 05),NSE含量显著减少(P <0. 05);纹状体中SOD水平和GSH-Px水平显著增加(P <0. 05),MDA水平明显减少(P <0. 05),纹状体中TH表达和Bcl-2表达明显增加(P <0. 05),α-syn和Bax表达明显减少(P <0. 05);与美多巴组比较,芍药甘草汤加减高剂量组的行为学实验中游泳实验得分和悬挂实验得分明显增加(P <0. 05),爬杆时间明显减少(P <0. 05),血清中Cys-C含量明显增加(P <0. 05),而NSE含量明显减少(P <0. 05);纹状体中SOD水平和GSH-Px水平明显增加(P <0. 05),MDA水平明显减少(P <0. 05),纹状体中TH和Bcl-2表达显著增加(P <0. 05),α-syn和Bax表达明显减少(P <0. 05)。结论:芍药甘草汤加减对MPTP诱导的帕金森病模型小鼠神经保护作用可能是通过抑制氧化损伤和细胞凋亡来实现的。

Objective: To study the effect of modified Shaoyao Gancao Tang on 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine( MPTP)-induced Parkinson's disease model mice. Method: The sixty C57/BL6 mice were randomly divided into normal group,model group,Madopar group( 50 mg·kg^-1) and low,medium and high doses modified Shaoyao Gancao Tang( 1,2,4 g·kg^-1). The modeling method was to intraperitoneally inject C57/BL6 mice with MPTP( 40 mg·kg^-1) once a day for 7 days. Except normal group and model group were given normal saline daily,drug-administered group was intragastrical administered once a day,and the third day after the drug was administered according to the above method( except normal group). The behavior of each group of mice( climbing test,swimming test,and tail suspension test) was examined up to 15 days after administration.Subsequently,the levels of Cystatin-C( Cys-C) and neuron-specific enolase( NSE) in the serum of each group were detected by enzyme-linked immunosorbent assay( ELISA). The levels of superoxide dismutase( SOD),malondialdehyde( MDA) and glutathione peroxidase( GSH-Px) in striatum of each group were detected by spectrophotometry. The protein expression of α-synuclein(α-syn),tyrosine hydroxylase( TH),B lymphoma-2 gene( Bcl-2) and Bcl-2 related X protein( Bax) in the striatum of each group was detected by Western blot.Result: Compared with normal group,the swimming experiment scores and suspension experiments scores in the behavioral experiments of model group mice were significantly lower( P<0. 01), The climbing time was significantly increased( P<0. 01);the serum Cys-C content was significantly decreased( P<0. 01),serum NSE content increased significantly( P<0. 05). SOD level and GSH-Px level in striatum decreased significantly( P<0. 01),MDA level in striatum increased significantly( P<0. 05),TH and Bcl-2 expression in striatum were significantly decreased( P<0. 01),α-syn and Bax expression in striatum were significantly increased( P<0. 05). Compared with model group,the scores of swimming experiment and suspension experiment in the behavioral experiments of Shaoyao Gancao Tang group and Madopar group were significantly increased( P<0. 05),while the climbing time was significantly reduced( P<0. 05). The serum Cys-C content increased significantly( P<0. 05),the serum NSE content decreased significantly( P<0. 05). The SOD level and GSH-Px level in the striatum increased significantly( P<0. 05),while the MDA level in the striatum decreased significantly( P<0. 05). There was also a significant increase in TH and Bcl-2 expression in the striatum( P<0. 05),α-syn and Bax expression in striatum were significantly decreased( P<0. 05). Compared with Madopar group,the scores of swimming experiment and suspension experiment in the behavioral experiment of high-dose group of Shaoyao Gancao Tang were significantly increased( P<0. 05),while the climbing time was significantly reduced( P<0. 05). The serum Cys-C content increased significantly( P<0. 05),while the serum NSE content decreased significantly( P<0. 05). The SOD level and GSH-Px level in the striatum increased significantly( P<0. 05),while the MDA level in the striatum decreased significantly( P<0. 05). There was also a significant increase in TH expression and Bcl-2 expression in the striatum( P<0. 05),while α-syn expression and Bax expression in the striatum were significantly decreased( P<0. 05). Conclusion: The neuroprotective effect of modified Shaoyao Gancao Tang on MPTP-induced Parkinson's disease model mice may be achieved by inhibiting oxidative damage and apoptosis.