淀粉样β-蛋白1-42和thiorphan对恒河猴海马结构的影响  

Effects of intracranial co-infusion of Aβ_(1-42) and thiorphan on Macaca Rhesus hippocampal formation

作　　者：李文德[1] 吴玉娥[2] 闵凡贵[1] 李卓[3] 黄家园[4] 黄韧[2] 

机构地区：[1]广东天然药物研究与开发重点实验室,广东医学院药理学教研室,广东湛江524023 [2]广东省实验动物监测所,广东广州510260 [3]广东医学院附属医院,广东湛江524023 [4]广东医学院第一临床学院,广东湛江524023

出　　处：《中国药理学通报》2010年第2期186-190,共5页Chinese Pharmacological Bulletin

基　　金：广东省科技计划资助项目(No2006B36004015,2009B060300017);广东省自然科学基金资助项目(No010086,06023013)

摘　　要：目的观察淀粉样β蛋白1-42(Aβ1-42)和thiorphan对恒河猴大脑海马神经元淀粉样蛋白、乙酰胆碱转移酶(ChAT)以及胶质纤维酸性蛋白(GFAP)表达的影响,为注射thior-phan和Aβ1-42建立恒河猴AD模型提供数据准备。方法开颅后先注射thiorphan到猕猴的大脑皮质和基底核,消耗已存在的Neprilysin,然后再缓慢的注射孵育好的纤丝状Aβ1-42,后植入含有thiorphan的微渗透泵到基底核。HE染色观察海马结构病理改变。免疫组化的方法检测猴子海马Aβ1-42、乙酰胆碱转移酶(ChAT)、胶质纤维酸性蛋白(GFAP)阳性细胞的表达。结果HE染色显示海马CA1、CA2等区神经元萎缩,海马齿状回局部颗粒细胞丢失被胶质细胞取代;免疫组化结果显示模型组海马各区Aβ1-42、ChAT、GFAP阳性细胞吸光度(OD值)分别为0.59±0.05、0.21±0.04、0.19±0.04,与对照组比较P<0.01,差异具有统计学意义。结论Aβ1-42和thiorphan联合颅内给药导致恒河猴海马产生类似AD患者的病理改变。Aim To observe the change of amyloid, acetylcholine transferase and glial fibrillary acidic protein expression in Macaca Rhesus hippocampal after infused the Aβ1-42 and thiorphan and explore the possibility of the establishment of Macaca Rhsus AD model in brain. Method The Rhesus monkeys were anesthetized （ira）, the skull was exposed by a midline scalp incision, and oriented craniotomy was performed on leftside by dental drill. First, neprilysin in cerebral cortex and basal nucleus was consumed by infusion thiorphan. Then cerebral cortex and basal nucleus were slowly infused with fibrilla Aβ1-42. Finally, the cannula for thiorphan infusion was implanted into the basal nucleus. Miniosmotic pump （ Alzet MODEL 2ML4, ） was subcutaneously fixed by bio gel 454 on the calvaria （Loetite Co. Ltd,USA） according to the manufacturer＇s instructions. After 50 days＇ survival, animals were deep anesthetized with ketamine and sacrificed. The pathological changes were observed by HE staining and immunostaining in monkey brains. Result Neuronal loss and a proliferation of microglia were detected in hippocampal formation by HE staining. Immuno-staining showed Aβ1-42 ChAT and GFAP positive ceils density were 0. 59 ±0. 05,0. 21±0. 04 and 0. 19 ±0. 04 separately. Compared with control group, the density in ex-perimental groups showed distinct difference in statistic analysis （P 〈 0.01 ）. Conclusion The same pathological change was detected in the thioaphan and Aβ1-42 infusion in Macaca Rhesus hippocampal formation as what was found in AD patients.

关 键 词：阿尔采末病 恒河猴 淀粉样β蛋白1-42 THIORPHAN 乙 酰胆碱转移酶 胶质纤维酸性蛋白 免疫组化 

分 类 号：R-332[医药卫生] R322.81