特质与状态焦虑动物模型相关性及中药有效组分干预

"Correlation between trait and state anxiety in animal models" is an unsolved scientific problem in Neuropsychopharmacology and Behavioral Pharmacology. There are always several defects, such as the heterogeneity and resistance, in the classic state anxiety animal models alone, open field test(OFT), elevated plus maze(EPM), light/dark box(LDB). Modern pharmacology has proved that GABAA/benzodiazepine receptor complex is the main target for anxiolytic and anxiogenic compounds in both animal model behavioral research and clinical therapeutics. Anxiolytic-like effect of Paeonol, the major component of a traditional Chinese herb(Danpi), has been confirmed by behavioral pharmacological analysis, while the mechanism is unclear, although it is in full compliance with the hypothetic structural model of phenol derivatives for direct activation of chloride currents via GABAA receptor(GABAAR) that only compounds with (a) the unsubstituted phenolic hydroxyl attached directly to the benzene ring and (b) a methyl or isopropyl group inserted in ortho position to the phenolic hydroxyl were able to induce chloride inward currents via GABAAR in the absence of GABA. In order to verify the correlation between trait and state anxiety in animal models, novel animal models of anxiety, such as triple integrated apparatus(TIA, OFT+EPM+LDB), graded anxiety test(GAT, EPM+LDB)and free exploratory paradigm(FEP), are used to evaluate the emotional behaviors in Kunming mice. Whole-cell patch clamp for the GABAA/benzodiazepine receptor complex will be performed to explore the GABAergic contribution to the pharmacological action of Paeonol. Thus, the present study may reveal the anxiolytic-like mechanism of Paeonol, and give an answer to the question, that is there correlation between trait and state anxiety in animal models

"特质与状态焦虑动物模型相关性"一直是精神神经药理学和行为药理学学科悬而未解的科学问题，而经典状态焦虑动物模型（OFT，EPM，LDB）总是存在异质性及耐受性等诸多问题。现代药理学研究证实：抗焦虑药物主要作用靶点是γ-氨基丁酸A受体（GABAAR）；中药牡丹皮有效组分（丹皮酚）具有抗焦虑作用，丹皮酚及其主要代谢产物均符合"GABAAR配体结构法则"，其抗焦虑作用机制尚不明确。本研究采用新型焦虑动物模型（TIA，GAT），初步探讨"特质与状态焦虑动物模型相关性"科学问题；同时采用宏观行为药理学和微观受体药理学相结合方法，评价丹皮酚对特质与状态焦虑动物模型行为学、神经元细胞活力及GABAAR介导氯离子通道的影响，初步探讨中药牡丹皮有效组分丹皮酚抗焦虑作用的药理机制。

本项目围绕“特质与状态焦虑动物模型相关性”科学问题，按照原定计划开展经典焦虑动物模型旷场试验（OFT）、高架十字迷宫实验（EPM）、高架〇迷宫实验（EZM）、明暗箱实验（LDB）、自由探索模型（FEP）等方法学研究，并结合“行为组学”—整合模式，构建新型复合实验装置（mTIA），初步构建基于“行为组学”模式焦虑动物模型评价技术体系；采用行为药理学方法评价丹皮酚抗焦虑作用机制，借助MTT和LDH评价丹皮酚神经保护及细胞毒作用，利用膜片钳技术评价丹皮酚对GABAAR变构调节作用，初步阐释丹皮酚抗焦虑作用与GABAAR相关微观机制。本项目先后培养博士研究生1名，本科专题实习学生4名；申请前期发表相关学术论文10篇，执行期间发表相关学术论文6篇，会议论文3篇，其中SCI收录2篇，F5000收录1篇；先后荣获中国药理学会药源性疾病专业委员会第八届药源性疾病与安全用药中国论坛-老年人群分论坛一等奖、2015年岐黄奖第六届全国中医药博士生优秀论文二等奖、2015年全国博士生学术论坛（药学）纪念奖、2014年中国药学大会暨第十届中国药师周论坛优秀奖、2013年齐鲁研究生学术论坛-医学分论坛优秀学术论文一等奖等奖励。本项目依据当前焦虑动物模型最新研究进展，创造性提出mTIA（OFT+mEZM+LDB），其相比TIA（OFT+EPM+LDB）增加了二选式通路，更符合心理学特质，可以作为Battery模式和TIA的替代，也丰富了行为药理学焦虑动物模型内容；同时发现丹皮酚抗焦虑作用与变构调节GABAAR氯离子通道相关，丰富了受体药理学GABAAR配体模型内容。