五子衍宗丸对帕金森病小鼠神经元运动功能的作用及其可能的机制  

Effect and possible mechanism of Wuzi Yanzong Pill on motor function of neurons in Parkinson's disease mice

作　　者：潘涛 肖琪 樊慧杰 徐磊 贾璐 秦劭晨 王利然 马存根 张波 柴智 Pan Tao;Xiao Qi;Fan Hui-Jie;Xu Lei;Jia Lu;Qin Shao-Chen;Wang Li-Ran;Ma Cun-Gen;Zhang Bo;Chai Zhi(Key Research Laboratory of Benefiting Qi for Acting Blood Circulation Method to Treat Multiple Sclerosis of State Administration of Traditional Chinese Medicine/Neurobiology Research Center,Shanxi University of Chinese Medicine,Jinzhong,Shanxi 030619,China;The First Clinical College of Shanxi University of Chinese Medicine,Taiyuan,Shanxi 030024,China;The Second Clinical College of Shanxi University of Chinese Medicine,Taiyuan,Shanxi 030012,China)

机构地区：[1]山西中医药大学多发性硬化益气活血重点研究室/神经生物学研究中心,山西晋中030619 [2]山西中医药大学第一临床学院,山西太原030024 [3]山西中医药大学第二临床学院,山西太原030012

出　　处：《解放军医学杂志》2024年第5期550-556,共7页Medical Journal of Chinese People's Liberation Army

基　　金：国家中医药管理局青年岐黄学者培养项目[国中医药人教函(2022)256号];山西省重点国别科技合作项目(202204041101002);山西省青年拔尖人才支持计划项目(晋组办字[2019]35号文);山西省省筹资金资助回国留学人员科研项目(2021-142)。

摘　　要：目的观察五子衍宗丸(WYP)对帕金森病模型小鼠运动功能的作用及其可能的机制。方法24只雄性C57BL/6小鼠随机分为对照组、模型组和WYP组,每组8只。模型组及WYP组小鼠连续7 d腹腔注射1-甲基-4-苯基-1,2,3,6-四氢吡啶,建立帕金森病模型。WYP组小鼠从模型制备第1天开始,连续14 d采用WYP药液[16 g/(kg.d)]灌胃,2次/d;对照组和模型组同一时间用0.9%氯化钠溶液[50 ml/(kg.d)]灌胃,2次/d。步态实验检测各组小鼠行为学表现;免疫荧光染色检测各组小鼠黑质区酪氨酸羟化酶(TH)阳性细胞数,核因子E2相关因子2(Nrf2)的荧光强度和NeuN神经元与Nrf2a共染细胞数;Western blotting检测各组小鼠脑组织TH、Kelch样ECH相关蛋白1(Keap-1)、Nrf2、血红素加氧酶-1(HO-1)的表达水平。结果步态实验结果显示,与对照组比较,模型组小鼠左前爪、右前爪、左后爪、右后爪站立时间均明显缩短(P<0.01),左前爪、右前爪、左后爪、右后爪摆动时间均明显延长(P<0.05);与模型组比较,WYP组小鼠左前爪、右后爪站立时间延长(P<0.05),左前爪、右前爪摆动时间缩短(P<0.05)。免疫荧光染色和Western blotting检测结果显示,与对照组比较,模型组TH阳性细胞数与蛋白、Nrf2平均荧光强度与蛋白、HO-1水平降低(P<0.01),Keap-1水平增高(P<0.01),NeuN神经元与Nrf2共染细胞数降低(P<0.001);与模型组比较,WYP组小鼠脑组织TH阳性细胞数与蛋白、Nrf2平均荧光强度与蛋白、HO-1水平、NeuN神经元与Nrf2共染细胞数均增高(P<0.05),Keap-1蛋白水平降低(P<0.05)。结论五子衍宗丸可缓解帕金森病小鼠运动功能障碍,保护多巴胺能神经元,其作用机制可能与调控Keap-1/Nrf2/HO-1通路抑制氧化应激反应有关。Objective To observe the effects of Wuzi Yanzong Pill(WYP)on motor function in a mouse model of Parkinson's disease(PD)and to explore its potential mechanisms.Methods Twenty-four male C57BL/6 mice were randomly divided into control group,model group and WYP group,with 8 mice in each group.Mice in model and WYP group were intraperitoneally injected with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine for 7 consecutive days to establish a PD model.From the 1st day of model preparation,mice in WYP group were gavaged with WYP solution[16 g/(kg·d)]twice daily for 14 consecutive days.At the same time,mice in control group and model group were gavaged with 0.9%NaCl solution[50 ml/(kg·d)]twice a day.Gait experiment was utilized to assess the behavioral performance of mice in each group.Immunofluorescence staining was conducted to detect the number of tyrosine hydroxylase(TH)-positive cells in the substantia nigra region,the fluorescence intensity of nuclear factor E2-related factor 2(Nrf2),and the number of NeuN neurons co-labeled with Nrf2 in each group.Western blotting was employed to determine the expression levels of TH,Kelch-like ECH-associated protein 1(Keap-1),Nrf2,and heme oxygenase-1(HO-1)in the brain tissue of mice in each group.Results The gait experiment results showed that,compared with control group,standing time of the left front paw,right front paw,left hind paw,and right hind paw of the mice in model group was significantly shortened(P<0.01),while swinging time of the left front paw,right front paw,left hind paw,and right hind paw was significantly prolonged(P<0.05).Compared with model group,standing time of the left front paw and right hind paw of the mice in WYP group was significantly prolonged(P<0.05),while swing time of the left front paw and right front paw was significantly shortened(P<0.05).Immunofluorescence staining and Western blotting results showed that,compared with control group,in model group the number of TH-positive cells,average fluorescence intensity of Nrf2,and HO-1 levels decreased(P

关 键 词：五子衍宗丸 帕金森病 Keap-1/Nrf2/HO-1通路 神经元 

分 类 号：R742.5[医药卫生—神经病学与精神病学] R285[医药卫生—临床医学]