基于nNOS/CAPON相互作用探讨参枝苓口服液对拟散发性AD小鼠海马突触的影响  

Effects of Shenzhiling oral liquid on the interaction between neuronal nitric oxide synthase and carboxy-terminal PDZ ligand of neuronal nitric oxide synthase in the hippocampal synapses in a mouse model of sporadic Alzheimer’s disease

作　　者：田梅静 高晨艳 郑明翠 和雁南 王蓬文[1] TIAN Meijing;GAO Chenyan;ZHENG Mingcui;HE Yannan;WANG Pengwen(Key Laboratory of Chinese Internal Medicine of Ministry of Education and Beijing,Dongzhimen Hospital,Beijing University of Chinese Medicine,Beijing 100700,China)

机构地区：[1]北京中医药大学东直门医院中医内科学教育部和北京市重点实验室,北京100700

出　　处：《中国比较医学杂志》2022年第8期1-10,共10页Chinese Journal of Comparative Medicine

基　　金：国家自然基金面上资助项目(81573927);2019年度北京中医药大学青年教师项目(2019-BUCMXJKY018)。

摘　　要：目的探索参枝苓口服液对拟散发性阿尔茨海默病(sporadic Alzheimer’s disease,SAD)模型小鼠学习记忆能力的影响以及对神经元型一氧化氮合酶(neuronal nitric oxide synthase,nNOS)与神经元型一氧化氮合酶羧基末端PDZ配体(carboxy-terminal PDZ ligand of neuronal nitric oxide synthase,CAPON)相互作用及下游细胞外信号调控激酶(extracellular signal-regulated kinases,ERK)/环磷腺苷反应元件结合蛋白(cAMP-response element binding protein,CREB)信号通路的调节。方法72只C57BL/6J雄性小鼠随机抽取12只作为对照组,其余60只隔日双侧侧脑室注射链脲佐菌素(streptozotocin,STZ),制备拟SAD小鼠模型,对照组代以等体积人工脑脊液。造模组小鼠随机分为模型组、多奈哌齐组和参枝苓高、中、低剂量组,各治疗组给予相应剂量的药物干预,而对照组及模型组给予等体积0.5%CMC,连续灌胃3个月。Morris水迷宫记录小鼠的逃避潜伏期和撤台后穿越平台次数;透射电镜观察小鼠海马CA1区突触数量;免疫共沉淀检测小鼠海马组织中nNOS与CAPON的相互作用;免疫组织化学染色观察小鼠海马CA1区ERK、CREB的分布及阳性表达细胞数;Western blot检测小鼠海马ERK、p-ERK、CREB、p-CREB的表达。结果与对照组相比,模型组小鼠逃避潜伏期显著增加(P<0.01),穿越平台次数显著减少(P<0.01);海马CA1区突触数量显著减少(P<0.01);nNOS与CAPON的相互作用显著增加(P<0.01);ERK的表达无明显变化(P>0.05),p-ERK、CREB、p-CREB的表达显著下降(P<0.01),p-ERK/ERK、p-CREB/CREB水平显著降低(P<0.01)。与模型组相比,多奈哌齐组和参枝苓各干预组逃避潜伏期均明显下降(P<0.05或P<0.01),小鼠穿越平台次数均显著增加(P<0.01);海马CA1区突触数量均显著增多(P<0.01);nNOS与CAPON的相互作用均明显减少(P<0.05或P<0.01);ERK的表达无明显变化(P>0.05),p-ERK、CREB、p-CREB的表达均有不同程度的增加(P<0.05或P<0.01)),p-ERK/ERK、p-CREB/CREB水Objective To explore the effects of Shenzhiling oral liquid(SZL)on neuronal nitric oxide synthase(nNOS)and carboxy-terminal PDZ ligand of neuronal nitric oxide synthase(CAPON)interaction,and determine which downstream extracellular signal-regulated kinase(ERK)/cAMP-response element binding protein(CREB)signaling pathway regulates the learning and memory ability in a mouse model of sporadic Alzheimer’s disease(SAD).Methods Twelve of 72 C57 BL/6 J male mice were randomly selected as the control group.The remaining 60 mice had streptozotocin injected into the bilateral ventricles on the first and third days to create a model of SAD;the streptozotocin was replaced with an equal volume of artificial cerebrospinal fluid in the control group.The 60 SAD mice were randomly divided into the model group,donepezil group,high-dose SZL(SZL-H),medium-dose SZL(SZL-M)and low-dose SZL(SZL-L)groups.Each intervention group was gavaged with the corresponding drug for 3 months,whereas the control group and model group were gavaged with the same volume of 0.5%sodium carboxymethyl cellulose.The Morris water maze was used to test the escape latency and number of times each mouse crossed the platform after being removed from the platform.The number of synapses in the hippocampal CA1 region in each group was observed by transmission electron microscopy.The interaction between nNOS and CAPON in the hippocampus was detected by co-immunoprecipitation.The distribution and presence of ERK and CREB cells in the hippocampal CA1 region were observed by immunohistochemical staining.Western blot analysis was used to detect the expressions of ERK,p-ERK,CREB and p-CREB in hippocampal tissue.Results Compared with the control group,the model group had significantly increased escape latency(P<0.01),decreased frequency of platform crossing(P<0.01),decreased number of synapses in the hippocampal CA1 region(P<0.01),increased interaction between nNOS and CAPON(P<0.01),similar expression of ERK(P>0.05),decreased expressions of p-ERK,CREB and p-CREB(P<0.01),

关 键 词：散发性阿尔茨海默病 参枝苓口服液 神经元型一氧化氮合酶 神经元型一氧化氮合酶羧基末端PDZ配体 突触 

分 类 号：R-33[医药卫生]