安神定志丸及拆方对睡眠剥夺小鼠学习记忆及cAMP-CREB-BDNF信号通路蛋白表达的影响  

Effects of Anshen Dingzhi Pills and Its Compatibility on the Expressions of cAMP-CREBBDNF Signaling Pathway Protein,Learning and Memory Ability in Sleep Deprivation Mice

作　　者：曾顺 刘楠楠 徐波[2] 丁莉[2] ZENG Shun;LIU Nannan;XU Bo;DING Li(Taihe Hospital,Wuhan 430065,China;Hubei University of Chinese Medicine)

机构地区：[1]十堰市太和医院,湖北武汉430065 [2]湖北中医药大学

出　　处：《西部中医药》2022年第12期19-24,共6页Western Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金青年基金(81703972)。

摘　　要：目的:探讨安神定志丸及拆方对睡眠剥夺(sleep deprivation,SD)模型小鼠学习记忆水平及cAMP-CREB-BDNF蛋白表达的影响。方法:选取雄性昆明小鼠60只为研究对象,通过组方分析主要由安神药和化痰药组成,故拆方分为宁心安神组、补气化痰组,空白组、模型组、安神定志(原方)组和艾司唑仑组,每组10只。采用腹腔注射对氯苯丙氨酸(parachlorophenylalanine,PCPA)混悬液制备睡眠剥夺模型。造模后采用Morris水迷宫检测学习记忆水平;免疫组织化学法(immunohistochemistry,IHC)计算海马磷酸化环磷腺苷效应元件结合蛋白(p-cAMP-response element binding protein,p-CREB)、脑源性神经营养因子(brainderived neurotrophic factor,BDNF)阳性细胞表达;尼氏染色观察SCN神经元尼氏小体变化;酶联免疫吸附法(ELISA)检测血清环磷酸腺苷(Cyclic adenosine monophosphate,cAMP)、p-CREB、BDNF的浓度,蛋白质印迹法(Western blotting)检测海马中p-CREB、BDNF的蛋白水平。结果:与模型组比较,安神定志组上平台潜伏期、游泳总路程、第一次抵达平台时间缩短,跨越平台次数、目标象限时间增加,SCN神经元尼氏小体增加,cAMP、p-CREB、BDNF蛋白表达升高(P<0.01)。拆方结果显示,补气化痰组和宁心安神组均能够改善SD小鼠定位航行与空间探索能力,增加cAMP、p-CREB、BDNF蛋白表达,两组间比较差异有统计学意义(P<0.01,P<0.05),与安神定志组比较同时具有显著性差异(P<0.01,P<0.05)。结论:安神定志丸改善SD小鼠的学习记忆水平,与调节cAMP-CREB-BDNF信号通路蛋白表达有关,各组间影响程度依次为安神定志组>补气化痰组>宁心安神组。Objective:To explore the impacts of Anshen Dingzhi pills and its compatibility on the expressions of cAMP-CREB-BDNF signaling pathway protein,learning and memory ability in sleep deprivation(SD)mice.Methods:Sixty male Kunming mices were selected as the research objects,and divided into the groups of calming the heart and tranquilizing the mind,tonifying Qi and reducing phlegm,the blank group,the model group,Anshen Dingzhi(the original prescription)group and estazolam group,since the prescription is composed of the herbs of tranquilizing the mind and reducing phlegm,with ten mice in each group.Intraperitoneal injection of PCPA suspension was used to establish SD models.After modeling,Morris water maze was adopted to detect learning and memory ability;IHC was applied to calculate the expressions of hippocampal p-CREB,and positive cells of BDNF;nissl’s staining used to observe the changes of SCN neuron nissl bodies;ELISA method to detect the density of cAMP,p-CREB and BDNF,Western blotting to measure the levels of p-CREB and BDNF protein.Results:Compared with the model group,the incubation period of hopping on platform,total swimming distances,and the first arrival time on platform shortened in Anshen Dingzhi group,the number of crossing platforms,and target quadrant time increased,SCN neuron nissl bodies added,the expressions of cAMP,p-CREB and BDNF elevated(P<0.01).The results of the compatibility showed that the groups of calming the heart and tranquilizing the mind,tonifying Qi and reducing phlegm could improve place navigation and space exploration capability of SD mice,raise the expressions of cAMP,p-CREB and BDNF,and the difference had statistical meaning between both groups(P<0.01,P<0.05).Conclusion:Anshen Dingzhi pills could improve SD mice’s learning and memory ability,connected with regulating the expressions of cAMP-CREB-BDNF signaling pathway protein,the groups rank according to the degree of influence:Anshen Dingzhi group>tonifying Qi and reducing phlegm group>group of calming the heart and tranqui

关 键 词：睡眠剥夺 学习记忆 安神定志丸 神经因子 小鼠 动物实验 

分 类 号：R285.5[医药卫生—中药学]