新型化合物乙酰四氢小檗红碱的受体结合及抗焦虑效应(英文)  

Anxiolytic effect and receptor binding properties of acetylated tetrahydroberberrabine

作　　者：梁慧春 李诺敏 许宇辉[1] 叶恩茂[1] 张剑[3] 余伯阳[3] 杨征[1] 

机构地区：[1]军事医学科学院基础医学研究所认知与心理卫生研究中心,北京100850 [2]北京理工大学生命科学学院,北京100081 [3]中国药科大学中药复方研究室,江苏南京210009

出　　处：《中国药理学与毒理学杂志》2013年第6期931-940,共10页Chinese Journal of Pharmacology and Toxicology

基　　金：supported by National Science and Technology Major Projects of China(2008ZXJ09004-003)~~

摘　　要：目的 探讨延胡索四氢小檗碱同类物衍生物乙酰化四氢小檗红碱（THBr-A）的脑内受体结合特点及其抗焦虑效应。方法 ① 体外实验：制备稳定转染人肾上腺素α1，β1，多巴胺D2，多巴胺D3，γ-氨基丁酸（GABA）-A，GABA-B，N-甲基-D-天冬氨酸（NMDA），5-羟色胺1A（5-HT1A），5-HT2A受体基因的HEK293细胞膜，加入THBr-A 1×10-10 mol·L-1放射性配体结合实验观察THBr-A的脑内作用靶点。制备稳定转染人多巴胺D2受体基因的HEK293细胞膜，单独加入THBr-A 1×10-10~1×10-5 mol·L-1，或先加入喹吡罗10 μmol·L-1后再加入THBr-A 1×10-10~1×10-5 mol·L-1；或制备富含5-HT1A受体的SD大鼠海马组织细胞膜，单独加入THBr-A 1×10-10~1×10-5 mol·L-1[35S]GTP-γS实验观察D2和5-HT1A受体的内在活性。② 体内实验：雄性HaM/ICR小鼠按照分组分别于实验前60 min单次ig给予THBr-A 5，10，20，40和80 mg·kg-1，通过自发活动、孔板、高架十字迷宫及明暗穿箱实验观察THBr-A对小鼠自发活动和焦虑行为的影响。结果 ① 体外实验：放射性配体结合实验结果显示，THBr-A主要作用于D2，D3和5-HT1A受体，相应的百分抑制率分别为96.43%，77.61%和78.31%。[35S]-GTP-γS实验结果显示，THBr-A具有D2拮抗（IC50=11.54 nmol·L^-1）和5-HT1A部分激动（EC50=55.37 nmol·L-1）的内在活性。② 体内实验：自发活动实验结果显示，单次给予THBr-A 5~8 mg·kg^-1对小鼠给药后60 min内的自发活动数无影响（F（5，54）=1.247，P=0.300）。孔板实验结果显示，单次给予THBr-A 20，40和80 mg·kg^-1，小鼠的探头次数（F（6，63）=8.667，P〈0.01）和探头时间（F（6，63）=6.067，P〈0.01）明显增加。高架十字迷宫实验显示，单次给予THBr-A 20和40 mg·kg^-1，小鼠的入开臂时间百分比（F（6，63）=9.382，P〈0.01）和入开臂次数百分比（F（6，63）=8.957，P〈0.01）明显增加。明暗穿箱实验显示，单次给予THBr-AOBJECTIVE To investigate the brain receptor binding properties and the anxiolytic effects of acetylated tetrahydroberberrabine （THBr-A）, a new synthetic derivative of tetrahydroprotoberberines extracted from Chinese herbs Corydalis yanhusuo. METHODS ① In vitro The radio-ligand binding assays were performed by preparing the membrane of HEK293 cells stably transfected with the gene of human adrenergic α1, β1, dopamine D2, dopamine D3, γ-aminobutyric acid （GABA）-A, GABA-B, N-methyl-D-aspartic acid （NMDA）, serotonin receptor 1A （5-HT1A） and 5-HT2A, respectively, and then adding THBr-A 1×10-10 mol·L-1. The [35S]-GTP-γS assays were conducted by preparing the HEK293/D2 cell membrane and then adding THBr-A 1×10-10-1×10-5 mol·L-1 alone, or adding THBr-A 1×10-10-1×10-5 mol·L-1 following with quinpirole 10 μmol·L-1; or by preparing the rat hippocampal tissues cell membrane in which 5-HT1A receptors were highly expressed and then adding THBr-A 1×10-10-1×10-5 mol·L-1 alone, to investigate the intrinsic activity of THBr-A on D2 and 5-HT1A receptors. ②In vivo Naive HaM/ICR strain male mice were ig administered with a single dose of THBr-A 5, 10, 20, 40 and 80 mg·kg-1 60 min before test. Spontaneous activity, holeboard, elevated plus-maze, and light-dark transition test were used to assess the locomotor activity and anxiolytic effect of THBr-A. RESULTS ① In vitro The data of radio-ligand binding assays showed that THBr-A mainly interacted with D2, D3 and 5-HT1A receptors, and rate of inhibition for them was 96.43%, 77.61% and 78.31%, respectively. The results of [35S]-GTP-γS assays suggested that THBr-A exhibit both D2 antagonist （IC50=11.54 nmol·L-1） and 5-HT1A partial agonist （EC50=55.37 nmol·L-1） properties. ② In vivo The results of spontaneous activity test indicated that THBr-A after a single administration over the range 5 to 80 mg·kg-1 had no influence on the counts of mice locomotor activity in the 60 min （F（5,54）=1.247, P=0.300）. The results of holeboard test

关 键 词：抗焦虑 乙酰四氢小檗红碱 受体  多巴胺D2 受体  血清素  5-HT1 

分 类 号：R964[医药卫生—药理学]