复方地黄颗粒对帕金森病阴虚动风证大鼠纹状体PINK1、Parkin、α-syn表达的影响  

Effects of Compound Dihuang Granules on the Expressions of PINK1,Parkin andα-syn in Striatum of Rats with Parkinson Disease Yin-deficiency and Wind Movement Syndrome

作　　者：吕子微 梁建庆 康永强 孙光洁 王行玲 何建成[5] LYU Ziwei;LIANG Jianqing;KANG Yongqiang;SUN Guangjie;WANG Xingling;HE Jiancheng(School of Basic Medical Sciences,Gansu University of Chinese Medicine,Lanzhou 730000,China;Gansu Provincial Key Laboratory of Traditional Chinese Medicine Prescription Excavation and Innovation Transformation,Lanzhou 730000,China;Key Laboratory of Dunhuang Medicine and Transformation,Ministry of Education,Lanzhou 730000,China;The Second People's Hospital of Lanzhou City,Lanzhou 730000,China;School of Basic Medical Sciences,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China)

机构地区：[1]甘肃中医药大学基础医学院,甘肃兰州730000 [2]甘肃省中医方药挖掘与创新转化重点实验室,甘肃兰州730000 [3]敦煌医学与转化教育部重点实验室,甘肃兰州730000 [4]兰州市第二人民医院,甘肃兰州730000 [5]上海中医药大学基础医学院,上海201203

出　　处：《中国中医药信息杂志》2024年第8期117-125,共9页Chinese Journal of Information on Traditional Chinese Medicine

基　　金：国家自然科学基金(82160929);甘肃省科技厅第十一批省级科技计划自然科学基金(自筹经费)项目(21JR11RA143);敦煌医学与转化教育部重点实验室开放课题(DHYX18-08);甘肃中医药大学中医学一级学科“岐黄英才”导师专项基金硕导项目(ZYXKSD-202211)。

摘　　要：目的观察复方地黄颗粒对帕金森病(PD)阴虚动风证大鼠纹状体PTEN诱导假定激酶1(PINK1)、Parkin、α-突触核蛋白(α-syn)表达的影响,探讨其治疗PD阴虚动风证的作用机制。方法采用黑质注射6-羟基多巴胺制备PD阴虚动风证模型。将65只成模大鼠随机分为模型组、美多芭(150mg/kg)组和中药低、中、高剂量(1.75、3.5、7g/kg)组,另取26只大鼠分为正常对照组和假手术组,每组13只,各给药组分别予相应药液灌胃,正常对照组、假手术组和模型组予等体积生理盐水灌胃,每日1次,连续28d。检测大鼠行为学变化,ELISA和生化试剂盒分别检测纹状体三磷酸腺苷(ATP)含量及线粒体呼吸链复合物Ⅳ(COXⅣ)活性,HE染色观察纹状体组织形态,免疫组化染色、Westernblot和RT-qPCR检测纹状体PINK1、Parkin、α-syn蛋白和mRNA表达。结果与正常对照组和假手术组比较,模型组大鼠旋转圈数和爬杆时间增加,游泳评分降低(P<0.01),纹状体ATP含量及COXⅣ活性降低(P<0.01),神经元排列混乱、数量减少、细胞肿胀,部分细胞核皱缩,空泡形成,边界模糊,纹状体PINK1、Parkin蛋白和mRNA表达降低(P<0.01),α-syn蛋白和mRNA表达升高(P<0.01);与模型组比较,各给药组大鼠旋转圈数和爬杆时间减少,游泳评分升高(P<0.01),纹状体ATP含量及COXⅣ活性升高(P<0.05,P<0.01),神经元数量增加、排列趋于整齐、形态有所改善,纹状体PINK1、Parkin蛋白和mRNA表达升高(P<0.05,P<0.01),α-syn蛋白和mRNA表达降低(P<0.05,P<0.01),以美多芭组和中药高剂量组改善最明显(P<0.05,P<0.01),2组差异无统计学意义(P>0.05),且中药各剂量组呈剂量依赖性(P<0.05,P<0.01)。结论复方地黄颗粒可改善PD阴虚动风证大鼠行为学症状,可能通过调节纹状体PINK1、Parkin、α-syn表达,进而减轻线粒体功能障碍,保护多巴胺能神经元,发挥治疗PD作用。Objective To observe the effects of compound Dihuang Granules on striatum PINK1,Parkin andα-syn in rats with Parkinson disease yin-deficiency and wind movement syndrome;To explore the mechanism of compound Dihuang Granules in the treatment of Parkinson disease yin-deficiency and wind movement syndrome.Methods 6-hydroxydopamine was injected into the substantia nigra of the midbrain to prepare the model of PD yin-deficiency and wind movement syndrome.Totally 65 model rats were randomly divided into model group,Madopar(150 mg/kg)group,and TCM low-,medium-,and high-dosage(1.75,3.5,7 g/kg)groups.Another 26 rats were divided into normal control group and sham-operation group,with 13 rats in each group.Each administration group was given corresponding medication solution by gavage,the normal control group,sham-operation group and the model group were given equivalent volume of normal saline,once a day for 28 consecutive days.The behavioral changes of rats were detected,ELISA and biochemical assay kits were used to detect ATP content and COXⅣactivity in striatum,HE staining was used to observe the morphological changes of the striatum,immunohistochemistry staining,Western blot and RT-qPCR were used to detect the expression of PINK1,Parkin,α-syn protein and mRNA in striatal tissue.Results Compared with the normal control group and the sham-operation group,the rats in the model group showed increased number of rotations and pole climbing time,decreased swimming score(P<0.01),the ATP content and COXⅣactivity in striatum tissue decreased(P<0.01),with disordered arrangement and reduced number of neurons,cell swelling,partial nuclear shrinkage,formation of vacuoles,and blurred boundaries,the expressions of PINK1,Parkin protein and mRNA in striatal tissue decreased(P<0.01),and the expressions ofα-syn protein and mRNA increased(P<0.01).Compared with the model group,the number of rotations and pole climbing time of rats in each medication group decreased,swimming scores increased(P<0.01),ATP content and COXⅣactivity in s

关 键 词：复方地黄颗粒 帕金森病 阴虚动风证 线粒体 大鼠 

分 类 号：R285.5[医药卫生—中药学]