水木和宁方通过调控泛素-蛋白酶体通路对MPP+诱导帕金森病体外模型细胞损伤的影响  

Effect of Shuimu Hening Recipe(水木和宁方)on MPP+Induced Injury of Parkinson's Disease Model Cells in Vitro by Regulating Ubiquitin-proteasome Pathway

作　　者：张天琪[1] 李传成 邱朝阳 刘萍 翟茜茜 霍青[5] ZHANG Tian-qi;LI Chuan-cheng;QIU Zhao-yang;LIU Ping;Zhai Xi-xi;HUO Qing(Shandong University of Traditional Chinese Medicine,Jin'an Shandong 250355;Linyi Central Hospital,Linyi Shandong 276400,China;Weifang Medical University,Weifang Shandong 261000,China;Shandong Maternity and Child Health Hospital,Ji'nan Shandong 250014,China;Affiliated Hospital of Shandong University of Traditional Chinese Medicine,Ji'nan Shandong 250011,China)

机构地区：[1]山东中医药大学,山东济南250355 [2]临沂市中心医院,山东临沂276400 [3]潍坊医学院,山东潍坊261000 [4]山东省妇幼保健院,山东济南250014 [5]山东中医药大学附属医院,山东济南250011

出　　处：《中医药导报》2022年第3期7-12,共6页Guiding Journal of Traditional Chinese Medicine and Pharmacy

基　　金：山东省重点研发项目(2018GSF119021)。

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。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 exe

关 键 词：帕金森病 水木和宁方 泛素-蛋白酶体通路 细胞凋亡 PC12细胞 

分 类 号：R285.5[医药卫生—中药学]