天麻超微粉调控胆碱能系统改善血管性痴呆大鼠学习记忆能力  

Effect of Ultrafine Powder of Gastrodiae Rhizoma in Improving Learning and Memory Ability of Vascular Dementia Rats by Regulating Cholinergic System

作　　者：陈婷婷 周雪[1] 徐旖旎[1] 杨非 吴小盈 冉芮 向泉 梁斌 陶玲[1] 沈祥春[1] CHEN Ting-ting;ZHOU Xue;XU Yi-ni;YANG Fei;WU Xiao-ying;RAN Rui;XIANG Quan;LIANG Bin;TAO Ling;SHEN Xiang-chun(Key Laboratory of Optimal Utilization of Natural Medicine Resources,Key Laboratory for Natural Medicinal Pharmacology and Drugability E valuation for Instiutions of Higher Education in Guizhou Province,Guizhou Medical University,Guiyang 550025,China;Guizhou Weimen PharmaceuticalCo.Ltd,Guiyang 550025,China)

机构地区：[1]贵州医科大学天然药物优效利用重点实验室,贵州省高等学校天然药物药理与成药性评价特色重点实验室,贵阳550025 [2]贵州威门制药有限公司,贵阳550025

出　　处：《中国实验方剂学杂志》2020年第15期26-32,共7页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：贵州省天然药物成药性人才基地项目(黔人颁发[2018]3号);贵州省社会攻关计划项目(黔科合[2016]支撑2826);贵州省科技厅联合学术新苗项目(黔科合平台人才[2017]5718);贵州省科技厅联合学术新苗项目(黔科合平台人才[2018]5779-20)。

摘　　要：目的:天麻超微粉(UPG)是否可以改善血管性痴呆(VD)大鼠的学习记忆功能障碍并且延缓VD的进程,该作用是否与乙酰胆碱酯酶(AChE)和胆碱乙酰转移酶(ChAT)调节乙酰胆碱(Ach)的释放从而调控胆碱能系统有关。方法:将SD大鼠随机分出假手术组,其余大鼠通过永久性大脑中动脉闭塞法(pMCAO)复制VD大鼠模型,并随机分为模型组,天麻超微粉低剂量组(0.45 g·kg-1),天麻超微粉高剂量组(1.8 g·kg-1),石杉碱甲组(80μg·kg-1),每组各12只。给药组每天灌胃1次,持续8周,假手术组和模型组给予等量蒸馏水灌胃后,通过Morris水迷宫实验对各组大鼠的学习和记忆能力进行评估;苏木素-伊红(HE)染色对大鼠海马CA1区进行病理形态学观察;酶联免疫吸附测定(ELISA)检测大鼠海马组织中Ach的水平;蛋白免疫印迹法检测大鼠海马组织中AChE,ChAT的蛋白表达;免疫组化实验观察海马组织CA1区ChAT的表达情况。结果:与假手术组比较,模型组的逃避潜伏期增加,穿过平台的次数和在目标象限停留的时间显著减少(P<0.01),模型组海马CA1区神经细胞排列无序,细胞核固缩且胞浆颜色较深,坏死细胞数量增多,海马组织中Ach的水平显著下降(P<0.01),AChE蛋白表达水平显著上调(P<0.01),ChAT蛋白表达水平明显下调(P<0.05);与模型组比较,各给药组能够显著降低模型大鼠的逃避潜伏期,显著增加穿过平台的次数和在目标象限停留的时间(P<0.01),神经细胞排列趋于整齐,细胞核呈圆形且胞浆染色浅,坏死细胞减少,海马组织中Ach水平明显增加(P<0.05),AChE蛋白表达显著下调,ChAT蛋白表达显著上调(P<0.01)。结论:天麻超微粉能够改善血管性痴呆大鼠的学习记忆障碍,其作用机制与上调ChAT蛋白,抑制AChE蛋白,提高脑内乙酰胆碱的作用密切相关。Objective:To investigate whether ultrafine powder of Gastrodiae Rhizoma(UPG)can alleviate the learning and memory impairment of vascular dementia rats and delay the process of VD,and whether this effect is related to the release of acetylcholine(Ach)through the regulation with acetylcholinesterase(ACh E)and choline acetyltransferase(Ch AT)and control of cholinergic system.Method:SD rats were randomly divided into sham operation group,UPG low dose group(0.45 g·kg-1),UPG high dose group(1.8 g·kg-1)and Huperzine A group(80μg·kg-1),with 12 rats in each group.The drug administration groups were given orally drugs once a day for 8 weeks,and sham group and model group were given orally the same amount of distilled water.The learning and memory ability of the rats with VD were evaluated by the Morris water maze.Htoxylin eosin(HE)staining was used for pathomorphological observation of hippocampus CA1 area of the rats.The content of Ach was determined by enzyme-linked immunosorbent assay(ELISA),ACh E and Ch AT protein expressions were detected by Western blot,and expression of Ch AT in hippocampus CA1 area was observed by immunohistochemistry.Result:Compared with the sham operation group,the escape latency of the model group was significantly increased(P<0.01),and the frequency of crossings platform and the time of staying in the target quadrant were reduced significantly(P<0.01).HE staining of hippocampal tissues from VD rat showed neuron disorders,loss and degeneration and necrosis,pyknosis of the nucleus and light coloration of the cytoplasm.The level of acetylcholine in the hippocampus was significantly decreased by ELISA(P<0.05),the expression level of ACh E protein was significantly up-regulated,and the expression level of Ch AT protein was significantly down-regulated(P<0.01).Compared with model group,each administration group could significantly reduce the escape latency of the model rats,and significantly increase the frequency of crossing platform and the time of staying in the target quadrant(P<0.01),the con

关 键 词：天麻超微粉 血管性痴呆 学习记忆功能障碍 乙酰胆碱酯酶(AChE) 胆碱乙酰转移酶(ChAT) 

分 类 号：R2-0[医药卫生—中医学] R22