【摘要】 目的：探讨水木和宁方通过调控泛素-蛋白酶体通路（UPP）对帕金森病细胞模型的影响。方法：通过MPP+诱导帕金森病体外细胞模型，利用MTT法检测细胞活性，筛选美多芭、水木和宁方药物干预最佳浓度。药物干预后，采用Real-time PCR法和Western blotting法分别检测与UPP相关的泛素（UB）、泛素激活酶E1(UBE1)、Parkin、泛素羧基端水解酶-L1(UCH-L1)、α-突触核蛋白（α-syn）、酪氨酸羟化酶（TH）的mRNA和蛋白表达水平。结果：MTT法筛选MPP+最佳造模浓度为1mmol/L，水木和宁方、美多芭药物最佳浓度分别为6 mg/mL和3.125μg/mL。镜下观察显示水木和宁方、美多芭对MPP+诱导的PC12细胞具有保护作用，可以提高细胞存活率。Real-time PCR法和Western blotting结果表明，与对照组比较，模型组PC12细胞TH、UB、UBE1、Parkin、UCH-L1的mRNA和蛋白表达均明显下调（P<0.05）,α-syn的mRNA和蛋白表达均明显上调（P<0.05）；与模型组比较，水木和宁方组、美多芭组PC12细胞TH、UB、UBE1、Parkin、UCH-L1的mRNA和蛋白表达均明显上调（P<0.05）,α-syn的mRNA和蛋白表达均明显下调（P<0.05）；与美多芭组比较，水木和宁方组PC12细胞TH、UB、UCH-L1的mRNA和蛋白表达均明显上调（P<0.05）,α-syn的mRNA和蛋白表达明显下调（P<0.05）。结论：水木和宁方可能通过调控UPP功能促进异常蛋白降解，减少神经元凋亡，从而发挥抗帕金森病的作用。更多还原

【Abstract】 Objective: To investigate the effect of Shuimu Hening Recipe on Parkinson’s disease cell models by regulating the ubiquitin-proteasome pathway(UPP). Methods: MPP+ was used to induce Parkinson’s disease in vitro cell model, cell viability was detected by MTT method, and the optimal concentration of Madopar-Shuimu Hening Recipe were screened. After drug intervention, real-time polymerase chain reaction(RT-PCR)and Western blotting were used to detect the mRNA and protein expression levels of ubiquitin(UB) related to UPP, ubiquitin activating enzyme E1(UBE1), Parkin, ubiquitin carboxyl terminal hydrolase-L1(UCH-L1),α-synuclein(α-syn), and tyrosine hydroxylase(TH). Results: The optimal concentration of MPP+was 1 mmol/L by MTT method, and the optimal concentrations of Shuimu Hening Recipe-Madopar were 6 mg/m L and 3.125 μg/m L respectively. Microscopic observation showed that Shuimu Hening Recipe-Madopar had a protective effect on MPP+induced PC12 cells and could improve the cell survival rate. The results of RT-PCR and Western blotting showed that compared with the control group, the m RNA and protein expressions of TH, UB, Ub E1, Parkin and UCH-L1 in PC12 cells in the model group were significantly down-regulated(P<0.05), and the m RNA and protein expressions of α-syn were significantly up-regulated(P<0.05). Compared with the model group, the m RNA and protein expressions of TH, UB, Ub E1, Parkin and UCH-L1 in PC12 cells in Shuimu Hening Recipe group and Medopa group were significantly up-regulated(P<0.05), and the m RNA and protein expressions of α-syn were significantly down regulated(P<0.05). Compared with the Medopa group, the m RNA and protein expressions of TH, Ub and UCH-L1 in PC12 cells of Shuimu Hening Recipe group were significantly up-regulated(P<0.05),and the m RNA and protein expressions of α-syn were significantly down regulated(P <0.05). Conclusion:Shuimu Hening Recipe may promote the degradation of abnormal proteins and reduce neuronal apoptosis by regulating the function of UPP, thereby exerting an anti-Parkinson’s disease effect.更多还原