补肾止颤方对帕金森病小鼠模型神经保护及抗兴奋性氨基酸作用  

Effect of Bushen Zhichan Prescription on neuroprotection and anti-excitatory amino acid in mice with Parkinson’s disease

作　　者：刘蕾蕾 田心韵 李文涛[1] LIU Lei-lei;TIAN Xin-yu;LI Wen-tao(Shanghai Municipal Hospital of Traditional Chinese Medicine,Shanghai University of Traditional Chinese Medicine,Shanghai 200000,China)

机构地区：[1]上海中医药大学附属市中医医院,上海200000

出　　处：《中草药》2023年第21期7096-7103,共8页Chinese Traditional and Herbal Drugs

基　　金：上海市科委自然基金项目(22ZR1459300);上海市十三五重点专科建设项目(shslczdzk04901);上海市中医医院未来计划项目(WLHBBD-2021002K)。

摘　　要：目的研究补肾止颤方对帕金森病小鼠模型神经保护及抗兴奋性氨基酸作用。方法采用1-甲基-4-苯基-1,2,3,6-四氢吡啶盐酸盐(1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine,MPTP)诱导帕金森病小鼠模型,随机分为对照组、模型组、金刚烷胺(41.1 mg/kg)组和补肾止颤方低、中、高剂量(6.6、9.9、19.8 g/kg)组,每组8只。连续14 d给药干预后,通过爬杆和自主活动实验评价小鼠行为学,并选择补肾止颤方最佳作用剂量用于后续研究;苏木素-伊红(HE)染色观察脑组织病理变化;ELISA法测定外周血谷氨酸含量;Western blotting和qRT-PCR法检测小鼠黑质纹状体酪氨酸羟化酶(tyrosine hydroxylase,TH)、多巴胺转运体(dopamine transporter,DAT)、兴奋性氨基酸转运体1(excitatory amino acid transporter 1,EAAT1)、YY-1、β-catenin蛋白和TH、DAT、EAAT1、YY-1 m RNA表达。结果与模型组比较,补肾止颤方组小鼠行为学症状显著改善(P<0.05、0.001),纹状体区域细胞整体形态与排列情况得到改善,黑质纹状体TH、DAT表达显著升高(P<0.05、0.01、0.001),外周血谷氨酸含量显著降低(P<0.001),黑质纹状体YY1表达水平均显著降低(P<0.01、0.001),EAAT1表达水平均显著升高(P<0.05、0.001),β-catenin蛋白表达水平显著升高(P<0.05)。结论补肾止颤方对帕金森病小鼠多巴胺能神经元有一定保护作用,其机制可能与通过转录调控EAAT1表达,从而减少过量的谷氨酸有关。Objective To study the effect of Bushen Zhichan Prescription(补肾止颤方)on neuroprotection and anti-excitatory amino acid in mice model with Parkinson’s disease.Methods A mouse model of Parkinson’s disease was induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine hydrochloride(MPTP).The mice were randomly divided into control group,model group,amantadine hydrochloride(41.1 mg/kg)group,and Bushen Zhichan Prescription low-,medium-,high-dose(6.6,9.9,19.8 g/kg)groups,with eight mice in each group.After 14 consecutive days of administration intervention,the behavior of mice was evaluated through pole climbing and autonomous activity experiments,and the optimal dosage of Bushen Zhichan Prescription was selected for subsequent research;Hematoxylin eosin(HE)staining was used to observe pathological changes in brain tissue;ELISA method was used to measure the content of glutamate in peripheral blood;Western blotting and qRT-PCR methods were used to detect tyrosine hydroxylase(TH),dopamine transporter(DAT),excitatory amino acid transporter 1(EAAT1),YY-1,β-catenin protein and TH,DAT,EAAT1,YY-1 mRNA expressions in nigrostriatal tissue of mice.Results Compared with model group,the behavioral symptoms of mice in Bushen Zhichan Prescription group were significantly improved(P<0.05,0.001),the overall morphology and arrangement of cells in striatum area were improved,the expressions of TH and DAT in nigrostriatal tissue were significantly increased(P<0.05,0.01,0.001),glutamine content in peripheral blood was significantly reduced(P<0.001),YY1 expression level in nigrostriatal tissue was significantly reduced(P<0.01,0.001),EAAT1 expression level was significantly increased(P<0.05,0.001),β-catenin expression level was significantly increased(P<0.05).Conclusion Bushen Zhichan Prescription has a certain protective effect on dopaminergic neurons in mice with Parkinson’s disease,and its mechanism may be related to reducing excessive glutamate through transcriptional regulation of EAAT1 expression.

关 键 词：补肾止颤方 帕金森病 神经保护 兴奋性氨基酸毒性 地黄苷D 腺苷 尿囊素 莫诺苷 马钱苷 2 3 5 4′-四羟基二苯乙烯-2-O-β-D-葡萄糖苷 大黄素 松果菊苷 毛蕊花糖苷 

分 类 号：R285.5[医药卫生—中药学]