白头翁皂苷组合物对帕金森病和阿尔茨海默病模型小鼠的改善作用  

Improvement of Pulsatilla saponin composition on Parkinson's disease and Alzheimer's disease in mice

作　　者：黄天衍 苑仁祎坤 何佳 韩珊[1,2] 李佶朗 梁晓伟 高翔 高红伟 HUANG Tianyan;YUAN Renyikun;HE Jia;HAN Shan;LI Jilang;LIANG Xiaowei;GAOXiang;GAO Hongwei(College of Pharmacy,Guangxi University of Chinese Medicine,Nanning 530020,China;Guangxi Engineering Technology Research Center for Choice Chinese Patent Drug and Ethnic Drug Development,Nanning 530020,China)

机构地区：[1]广西中医药大学药学院,广西南宁530020 [2]中药壮瑶药创新药物教育部工程研究中心,广西南宁530020

出　　处：《药物评价研究》2024年第7期1433-1443,共11页Drug Evaluation Research

基　　金：广西高校引进海外高层次人才“百人计划”项目(05018064)。

摘　　要：目的探讨白头翁皂苷B4和常春藤皂苷C(HSC,又名白头翁皂苷B5)质量比为4∶1的组合物对帕金森病和阿尔茨海默病模型小鼠的保护作用及机制。方法取60只健康雄性C57BL/6小鼠,随机抽取10只作为对照组,剩余50只小鼠ip1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)制备帕金森病(PD)模型,模型构建成功后随机分为模型组、左旋多巴胺片(阳性对照,30 mg·kg^(-1))组和白头翁皂苷B4+HSC高、中、低剂量(20、10、5 mg·kg^(-1))组,每组10只,ig给药14 d,每天1次,对照、模型组小鼠ig等量0.9%的氯化钠溶液。每2天观察动物震颤行为出现与消失的时间,于给药第7、14天展开行为学实验并记录评分;HE染色法观察脑组织黑质病理学变化;免疫组化检测脑组织α-突触核蛋白表达;实时荧光定量PCR(qRTPCR)检测小鼠脑组织多巴胺转运体(DAT)、酪氨酸羟化酶(TH)mRNA表达。将50只APP/PS1转基因小鼠随机分为模型组、盐酸多奈哌齐(阳性药,1.5 mg·kg^(-1))组和白头翁皂苷B4+HSC高、中、低剂量(20、10、5 mg·kg^(-1))组,取10只同背景APP/PS1转基因阴性小鼠作为对照组,每天1次,连续ig给药28 d,模型组和对照组ig等量0.9%的氯化钠溶液。给药结束后进行行为学实验并记录评分;ELISA法检测小鼠血清中炎症因子白细胞介素(IL-6、IL-1β)、肿瘤坏死因子-α(TNF-α)水平、环氧化酶2(COX-2)和氧化应激因子活性氧(ROS),试剂盒检测超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH-Px)水平。结果在PD模型小鼠中,与模型组比较,白头翁皂苷B4+HSC显著降低震颤麻痹评分(P<0.01、0.001),显著降低旷场实验评分(P<0.01、0.001),显著降低爬杆实验评分(P<0.01、0.001),显著降低悬挂实验评分(P<0.01、0.001),显著降低游泳实验评分(P<0.01、0.001),显著降低行为学实验综合评分(P<0.001),明显改善黑质体结构损伤,明显降低α-突触核蛋白表达,显著升高小鼠脑组织DAT和TH mRNA水平(P<0.05、0.01、0Objective To investigate the protective effects of a combination of Pulsatilla saponin B4 and hederagenin C(HSC,anemoside B5)with a quality ratio of 4∶1 on Parkinson's disease(PD)and Alzheimer's disease(AD)models in mice and the underlying mechanisms.Methods Sixty healthy male C57BL/6 mice were randomly selected,with 10 mice serving as the control group.The remaining 50 mice were intraperitoneally injected with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)to establish a PD model.After the model was successfully established,the mice were randomly divided into the model group,the positive control group(levodopa hydrochloride tablets,30 mg·kg^(-1)),and the Pulsatilla saponin B4+HSC high,medium,and low dose groups(20,10,and 5 mg·kg^(-1)),with 10 mice in each group.The mice were ig given the drugs for 14 d,once a day.The control and model groups were ig given an equal volume of 0.9%sodium chloride solution.The onset and disappearance times of tremor behavior were observed every two days.Behavioral experiments were conducted and scores were recorded on days 7 and 14 of the treatment.Hematoxylin and eosin(HE)staining was used to observe pathological changes in the substantia nigra of the brain tissue.Immunohistochemistry was used to detect the expression ofα-synuclein in the brain tissue.Quantitative real-time polymerase chain reaction(qRT-PCR)was used to detect the expression of dopamine transporter(DAT)and tyrosine hydroxylase(TH)mRNA in the brain tissue.50 APP/PS1 transgenic mice were randomly divided into the model group,the positive drug group(donepezil hydrochloride,1.5 mg·kg^(-1)),and the high,medium,and low dose groups of Pulsatilla saponin B4+HSC(20,10,and 5 mg·kg^(-1)),respectively.Ten APP/PS1 transgenic wild-type mice were taken as the control group.The mice were administered ig once a day for 28 d.The model group and the control group were given the same amount of 0.9%sodium chloride solution.After the administration,behavioral tests were conducted and the scores were recorded.The levels of infla

关 键 词：白头翁皂苷B4 白头翁皂苷B5 帕金森病 阿尔茨海默病 Α-突触核蛋白 脑组织损伤 神经元损伤 炎症因子 氧化应激 

分 类 号：R285.5[医药卫生—中药学]