ATP介导的嘌呤能信号在阿尔茨海默病相关结肠运动障碍中的作用  

Role of purinergic signaling mediated by ATP in Alzheimer's disease-associated colonic motility disorder

作　　者：林旭红[1] 王慧超[2] 郭俊玲[3,4] 房晓鹏[1] 张俊士[5] 郭俊楠 李玉霞[1] 杨瑞林[1] 李铁军[2] 刘建林[2] 

机构地区：[1]河南大学淮河医院检验科,转化医学中心,河南开封475000 [2]河南大学第一附属医院肾内科,河南开封475000 [3]河南大学淮河医院心内科,河南开封475000 [4]河南大学医学院,河南开封475000 [5]河南大学淮河医院神经内科,河南开封475000

出　　处：《中国病理生理杂志》2016年第12期2113-2124,共12页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.U1304802;No.81500430)

摘　　要：目的：探讨ATP介导的嘌呤能信号在阿尔茨海默病（Alzheimer’s disease,AD）相关结肠运动障碍中的作用及其相关的分子机制。方法：（1）临床试验：收集我院20例AD患者进行研究,放射免疫法测定血浆中胃动素（motilin,MTL）、胆囊收缩素（cholecystokinin,CCK）、血管活性肠肽（vasoactive intestinal peptide,VIP）和一氧化氮（nitric oxide,NO）水平;高效液相色谱法（high-performance liquid chromatography,HPLC）检测血浆三磷酸腺苷（adenosine triphosphate,ATP）水平,同时对患者进行神经心理学检查并积分。（2）动物实验：利用AD小鼠进行Morris水迷宫实验,评估空间学习记忆功能;放射免疫法测定血浆中MTL、CCK、VIP和NO水平,HPLC法检测血浆ATP水平;免疫组织化学法检测乙酰胆碱转移酶（choline acetyltransferase,ChAT）、VIP、一氧化氮合酶（nitric oxide synthase,NOS）和ATP合酶的变化;Western blot和免疫组化法检测P2Y受体表达水平的变化。（3）离体实验：离体器官浴槽系统观察P2Y受体激动剂α,β-亚甲基ATP（α,β-methylene ATP,α,β-MeATP）对自发性及电刺激诱导的结肠平滑肌收缩的影响,细胞内微电极技术观察α,β-MeATP对结肠平滑肌细胞膜电位的影响。结果：与对照组比较,AD患者血浆的MTL和CCK水平明显降低（P〈0.01）,NO和ATP水平显著升高（P〈0.05或P〈0.01）,VIP无明显变化。患者简明精神状态检查积分（MMSE）降低（P〈0.05）,AD评定量表-认知分量表（ADAS-Cog）积分、神经精神问卷（NPI）积分和AD协作研究日常能力量表（ADCS-ADL）积分均明显高于对照组（P〈0.01）。AD小鼠4-6 d逃逸潜伏期明显延长（P〈0.05）,空间探索能力明显降低（P〈0.05）,AD小鼠血浆中MTL和CCK水平明显降低（P〈0.01）,NO和ATP水平显著升高（P〈0.05或P〈0.01）,VIP无明显变化,AD小鼠结肠表达ATP合酶的水平明显上调（P〈0.05）,但ChAT、VIP和NOS无明显改变,AIM：To explore the role of purinergic signaling mediated by ATP in the Alzheimer＇s disease（ AD）-related colon motility disorder and its related molecular mechanisms.METHODS：（ 1） Clinical trials：AD patients in our hospital were collected and studied.Radioimmunoassay was used for the determination of plasma motilin（ MTL）,cholecystokinin（ CCK）,vasoactive intestinal peptide（ VIP） and nitric oxide（ NO）,and high-performance liquid chromatography（ HPLC） was applied to test the level of adenosine triphosphate（ ATP）.The patients were assessed by neuropsychology and scored accordingly.（ 2） In animal experiments,AD mice received Morris water maze test,and the spatial learning and memory function were evaluated.The plasma levels of MTL,CCK,VIP and NO were examined by radioimmunoassay,and the level of ATP was measured by HPLC.Choline acetyltransferase（ ChAT）,VIP,nitric oxide synthase（ NOS） and ATP synthase were detected by immunohistochemistry.Western blot and immunohistochemistry were used to detect the expression of P2Y receptor.（ 3） In vitro,organ bath was applied to observe the effect of α,β-methylene ATP（ α,β-MeATP）,an agonist of P2Y receptor,on both spontaneous and electrically evoked contraction of colonic smooth muscle strip,and the technique of intracellular microelectrode was applied to observe the effect of α,β-MeATP on the membrane potential of colonic smooth muscle cells.RESULTS：Compared with control group,the levels of MTL and CCK were decreased（ P〈0.01）,and the levels of NO and ATP were increased（ P〈0.05 or P〈0.01）,while the VIP level was not changed.Mini-Mental State Examination（ MMSE） score was decreased（ P〈0.05）,Alzheimer＇s Disease Assessment Scale-Cognitive Subscale（ ADAS-Cog） score,Neuropsychiatric Inventory（ NPI） score and Alzheimer＇s Disease Cooperative Study-Activities of Daily Living Scale（ ADCS-ADL） were all increased as compared with control group（ P〈0.01）.The 4 - 6 d escape latency of AP

关 键 词：P2Y受体 阿尔茨海默病 肠神经系统 结肠运动障碍 

分 类 号：R363[医药卫生—病理学] R749.16[医药卫生—基础医学]