水木和宁方对帕金森病模型小鼠泛素-蛋白酶体系统的影响

Effect of Shuimuhening Compound on Ubiquitin-Proteasome System (UPS) in Mice with Parkinson’s Disease Model

目的:基于泛素-蛋白酶体系统(UPS)研究水木和宁方对帕金森病(PD)小鼠的影响。方法:C57BL/6小鼠颈背部皮下注射鱼藤酮建立慢性PD小鼠模型。造模成功PD小鼠随机分为模型对照组、美多芭0. 15 g/kg组、水木和宁方15、30、60 g/kg组。治疗组每天灌胃给药2次,溶剂对照组和模型组给予等体积生理盐水,连续4 w。观察小鼠行为学改变;免疫组化检测中脑黑质α-突触核蛋白(α-syn)、酪氨酸羟化酶(TH)表达;WB和RT-PCR方法检测中脑黑质α-syn、TH及UPS相关的蛋白及mRNA表达。结果:与溶剂对照组比较,PD模型对照组小鼠前肢、后肢步长显著降低,游泳评分显著降低,中脑黑质TH阳性细胞数显著降低,α-syn阳性表达显著上调,α-syn蛋白及mRNA表达明显上调,TH、u BE1、Parkin、UCH-L1、Ubiquitin蛋白及mRNA表达明显下调(P <0. 05或P <0. 01)。与模型对照组比较,水木和宁方15 g/kg、30 g/kg、60 g/kg组小鼠行为学改变明显改善;中脑黑质TH阳性细胞数显著增加,α-syn阳性表达显著下调;TH、UPS相关蛋白及mRNA表达明显上调,α-syn蛋白及mRNA表达显著下调(P <0. 05或P <0. 01)。结论:水木和宁方可能通过调节UPS功能,促进α-syn的降解,从而改善PD小鼠的运动功能。

Objective: To study the effects of Shuimuhening compound on PD mice based on ubiquitin-proteasome system( UPS). Methods:C57 BL/6 mice were injected subcutaneously with rotenone at the back of the neck to establish a chronic PD mouse model. Successfully modeled PD mice were randomly divided into the model group,0. 15 g/kg Madopar group and Shuimuhening groups( 15 g/kg,30 g/kg,and 60 g/kg). The mice in the treatment group were orally administered twice a day,while those in the solvent control group and the model group were given the same amount of saline water,for 4 weeks. Behavior changes of mice were observed. Immunohistochemical method was used to detect the expression of α-syn and TH in substantia nigra. The expression of α-syn,TH and UPS related proteins and mRNA were detected by WB and RT-PCR. Results: Compared with the solvent control group,the step length of both fore limb and hind limb was significantly decreased in the model group,score in swimming was significantly decreased,TH positive expression in substantia nigra was decreased,while the positive expression of α-syn was increased,the expressions ofα-syn protein and mRNA were significantly up regulated. Also the expressions of TH、u BE1、Parkin、UCH-L1 and Ubiquitin protein were significantly down regulated in the model group. Compared with the model group,the behavior changes were improved in all doses of Shuimuhening groups,TH positive expression in substantia nigra was significantly increased,the positive expression of α-syn was decreased,the expressions of TH and UPS related proteins and mRNA were significantly up regulated,the expressions ofα-syn proteins and mRNA were significantly down regulated. Conclusion: Shuimuhening compound may improve the motor function of PD mice by regulating UPS function and promoting the degradation of α-syn.