APPSWE Tg2576小鼠脑屏障结构

doi:10.16098/j.issn.0529-1356.2018.05.002

摘要/Abstract

摘要：

目的探讨小鼠的血脑屏障（BBB）及血脑脊液屏障（BCSFB）的基本结构及其在阿尔茨海默病（AD）发生、发展过程中结构、功能及及其超微结构的改变。方法实验用动物采用APPSWE Tg2576鼠，分为APPSWE转基因阳性鼠（模型组）和同窝生野生型小鼠（对照组），每组各20只。饲养16个月后进行全身灌流固定, 开颅切取侧脑室室壁及其脉络丛组织。采用免疫荧光及透射电子显微镜技术观察BBB及BCSFB的超微结构，从而观察AD模型脑屏障的改变。结果 AD模型组与对照组相比较，血管密度明显降低；AD小鼠脑屏障正常结构受到损害，主要是脑血管内皮细胞（或脉络丛内皮细胞）之间的连接及其细胞器受损，脉络丛超微结构也出现明显变化，主要表现为细胞间隙增宽，细胞之间的黏附连接等连接结构也有部分碰坏，胞质内出现较多的囊泡状结构等。结论和正常鼠相比，AD鼠脑屏障受到一定的损害，可能致使脑屏障的转运机制出现相应的改变并影响脑内β-淀粉样蛋白（Aβ）的清除，脑屏障中存在的稳态机制，如其分泌物和受体介导的信号传导也可能出现改变，这些因素可能共同参与了AD的形成和进展。

关键词: 阿尔茨海默病, 血脑屏障, 血脑脊液屏障, 脉络丛, 增殖, 血管神经单元, 免疫荧光, 小鼠

Abstract:

Objective To investigate the basic structure of the blood brain barrier (BBB) and blood cerebrospinal fluid barrier (BCSFB) in mice and their changes in structure, function, and ultrastructure during the development and progression of AD. Methods The APPSWE Tg2576 mice were used and divided into APPSWE transgenic positive mice (model group) and littermates wild type mice (control group), twety mice in each group. After 16 months of feeding, whole body perfusion was performed and the craniotomy was performed to obtain the lateral ventricle wall and its choroid plexus. Immunofluorescence and transmission electron microscopy were used to observe the ultrastructure of BBB and BCSFB, so as to observe the changes of brain barrier of AD model. Results The vascular density was significantly lower in the AD model group than in the control group; the normal structure of the brain barrier in AD mice was impaired, mainly due to the connection between the brain vascular endothelial cells (or choroid plexus endothelial cells) and their organelles being damaged. The ultrastructure of the choroid plexus also showed significant changes. The main manifestations were the widening of the intercellular space, and some of the connecting structures between the cells, such as adhesion and connection, and some vesicle-like structures in the cytoplasm. Conclusion Compared with normal mice, the brain barrier of AD rats is damaged, which may lead to corresponding changes in the brain barrier transport mechanism and affect the clearance of Aβ in the brain, and the steady-state mechanisms existing in the brain barrier, such as secretions and receptors thereof. Mediated signaling may also change, and these factors may be involved in the formation and progression of AD.

Key words: Alzheimer’s disease, Blood brain barrier, Blood cerebrospinal fluid barrier, Choroid plexus, Proliferation, Vascular nerve unit, Immunofluorescence, Mouse

崔占军刘芳赵凯冰李冰梅刘中华. APPSWE Tg2576小鼠脑屏障结构[J]. 解剖学报, 2018, 49(5): 571-578.

CUI Zhan-jun LIU Fang ZHAO Kai-bing LI Bing-mei LIU Zhong-hua. Brain barrier structure of APPSWE Tg2576 mice[J]. Acta Anatomica Sinica, 2018, 49(5): 571-578.

参考文献

［1］ Querfurth HW, LaFerla FM. Alzheimer’s disease［J］. N Engl J Med, 2010, 362（4）:329-344.

［2］ Alonso Vilatela ME, López-López M, YescasGómez P. Genetics of Alzheimer’s disease ［J］. Arch Med Res, 2012,43(8):622-631.

［3］ Kim DH, Yeo SH, Park JM, et al. Genetic markers for diagnosis and pathogenesis of Alzheimer’s disease［J］. Gene, 2014,545(2):185-193.

［4］ Steinberg S, Stefansson H, Jonsson T, et al. Loss-of-function variants in ABCA7 confer risk of Alzheimer’s disease［J］. Nat Genet, 2015,47(5):445-447.

［5］ Lambert JC, Ibrahim-Verbaas CA, Harold D, et al. Meta-analysis of 74046 individuals identifies 11 new susceptibility loci for Alzheimer’s disease［J］. Nat Genet, 2013,45(12):1452-1458.

［6］ Zuroff L, Daley D, Black KL,et al. Clearance of cerebral Aβ in Alzheimer’s disease: reassessing the role of microglia and monocytes［J］.Cell Mol Life Sci, 2017, 74(12): 2167-2201.

［7］ Abbott NJ, Patabendige AA, Dolman DE, et al. Structure and function of the blood-brain barrier［J］. Neurobiol Dis, 2010, 37(1):13-25.

［8］ Villeda SA, Luo J, Mosher KI, et al. The ageing systemic milieu negatively regulates neurogenesis and cognitive function［J］. Nature, 2011, 477(3762):90-94.

［9］ Liu YJ, Guo DW, Tian L, et al. Peripheral T cells derived from Alzheimer’s disease patients overexpress CXCR2 contributing to its transendothelial migration, which is microglial NF-alpha-dependent［J］. Neurobiol Aging, 2010, 31(2):175-188.

［10］Gosselet F, Saint-Pol J, Candela P, et al. Amyloid-beta peptides, Alzheimer’s disease and the blood-brain barrier［J］. Curr Alzheimer Res, 2013,10(10):1015-1033.

［11］Spector R, Keep RF, Robert SS, et al. A balanced view of choroid plexus structure and function: focus on adult humans［J］. Exp Neurol, 2015,267:78-86.

［12］Chalbot S, Zetterberg H, Blennow K, et al. Blood-cerebrospinal fluid barrier permeability in Alzheimer’s disease［J］. J Alzheimers Dis, 2011,25(3):505-515.

［13］Krzyzanowska A, Carro E. Pathological alteration in the choroid plexus of Alzheimer’s disease: implication for new therapy approaches［J］. Front Pharmacol, 2012,3:75.

［14］Silverberg GD, Mayo M, Saul T, et al. Alzheimer’s disease, normal-pressure hydrocephalus, and senescent changes in CSF circulatory physiology: a hypothesis［J］. Lancet Neurol, 2003,2(8):506-511.

［15］Gonzalez-Marrero Ⅰ, Gimenez-Llort L, Johanson CE, et al. Choroid plexus dysfunction impairs beta-amyloid clearance in a triple transgenic mouse model of Alzheimer’s disease［J］. Front Cell Neurosci, 2015,9:17.

［16］Begley DJ, Brightman MW. Structural and functional aspects of the blood-brain barrier［J］. Prog Drug Res, 2003, 61:39-78.

［17］Pardridge WM, Eisenberg J, Yang J. Human blood-brain barrier insulin receptor［J］. J Neurochem, 1985, 44(6):1771-1778.

［18］Zhang Y, Pardridge WM. Rapid transferrin efflux from brain to blood across the blood-brain barrier［J］. J Neurochem, 2001, 76(5):1597-1600.

［19］Zlokovic BV. The blood-brain barrier in health and chronic neurodegenerative disorders［J］. Neuron, 2008, 57(2):178-201.

［20］Daneman R. The blood-brain barrier in health and disease［J］. Ann Neurol, 2012,72(5):648-672.

［21］Zhong Z, Deane R, Ali Z, et al. ALS-causing SOD1 mutants generate vascular changes prior to motor neuron degeneration［J］. Nat Neurosci, 2008,11(4):420-422.

［22］Bell RD, Winkler EA, Sagare AP, et al. Pericytes control key neurovascular functions and neuronal phenotype in the adult brain and during brain aging［J］. Neuron, 2010, 68(3):409-427.

［23］Daneman R, Zhou L, Kebede AA, et al. Pericytes are required for blood-brain barrier integrity during embryogenesis［J］. Nature, 2010, 468(7323):562-566.

［24］Li F, Lan Y, Wang Y, et al. Endothelial Smad4 maintains cerebrovascular integrity by activating N-cadherin through cooperation with Notch［J］. Dev Cell, 2011,20(3):291-302.

［25］Dziegielewska KM, Ek J, Habgood MD, et al. Development of the choroid plexus［J］. Microsc Res Tech, 2001, 52(1):5-20.

［26］Saunders NR, Daneman R, Dziegielewska KM, et al. Transporters of the bloodbrain and blood-CSF interfaces in development and in the adult［J］. Mol Aspects Med, 2013, 34(2-3):742-752.

［27］Speake T, Whitwell C, Kajita H, et al. Mechanisms of CSF secretion by the choroid plexus［J］. Microsc Res Tech, 2001,52(1):49-59.

［28］de Lange EC. Potential role of ABC transporters as a detoxification system at the blood-CSF barrier［J］. Adv Drug Deliv Rev,2014,56(12):1793-1809.

［29］Bergen AA, Kaing S, ten Brink JB, et al. Gene expression and functional annotation of human choroid plexus epithelium failure in Alzheimer’s disease［J］. BMC Genomics, 2015,16:956.

［30］Serot JM, Béné MC, Foliguet B, et al. Morphological alterations of the choroid plexus in late-onset Alzheimer’s disease［J］. Acta Neuropathol, 2000, 99(2):105-108.

[1]	洪晓敏李三强崔钦奕郑润月杨孟利罗仁利李前辉 . 酒精性肝纤维化核因子κB信号通路与性别的差异 [J]. 解剖学报, 2024, 55(1): 55-61.
[2]	袁蕾杨智伟杨惠刘政海何洁万炜. 三七总皂苷抑制小鼠星形胶质细胞活化缓解完全弗氏佐剂所致炎性痛 [J]. 解剖学报, 2024, 55(1): 25-31.
[3]	马婷婷陈建红刘爱翠李海宁. 抑制lncRNA TUG1下调核苷酸结合寡聚结构域样受体蛋白1炎症小体在延缓阿尔茨海默病进展的作用 [J]. 解剖学报, 2024, 55(1): 32-42.
[4]	邹成功冯浩陈兵唐辉邵川孙谋杨荣何家全. 创伤性颅脑损伤中神经功能障碍与认知功能损伤的动态变化 [J]. 解剖学报, 2024, 55(1): 43-48.
[5]	苏芃王兴仪梁景岩熊天庆. 一种低密度及高纯度胎鼠原代海马神经元培养方法的优化及鉴定[J]. 解剖学报, 2024, 55(1): 113-119.
[6]	许亚平余悦欣陈金福王柯柯郭志坤 . 高龄大鼠心室肌间质弹性纤维和胶原纤维的结构分布特征及其机制 [J]. 解剖学报, 2023, 54(6): 716-721.
[7]	刘丽平朱梦妮徐慧 . 白细胞介素6对脂多糖诱导所致类子痫前期大鼠有核红细胞的影响 [J]. 解剖学报, 2023, 54(6): 722-729.
[8]	赵卫明李晓余国营王改平常翠芳 . 丝氨酸肽酶抑制因子Kazal型1对肝癌细胞增殖的影响及其机制 [J]. 解剖学报, 2023, 54(6): 695-702.
[9]	张琴杨俊霞蒋青松. 福辛普利对糖尿病视网膜病变小鼠血管紧张素转化酶2和血管内皮生长因子的影响[J]. 解剖学报, 2023, 54(6): 628-634.
[10]	陈子璇司丽娜舒薇菡张欣魏晨阳程露阳杨松鹤乔跃兵. 褪黑素调控下丘脑PI3K/Akt/mTOR信号延缓雌性小鼠青春期启动的机制 [J]. 解剖学报, 2023, 54(6): 644-651.
[11]	刘哲川李坤马帅男孟家琪王艳芹. 利拉鲁肽对百草枯诱导的小鼠帕金森病模型炎症及线粒体融合/分裂的影响 [J]. 解剖学报, 2023, 54(6): 676-681.
[12]	刘叶楚亚楠徐岑璐何佳澄苏炳银太颢然. Roscovitine挽救帕金森病小鼠相关脑区神经元丢失和神经炎症[J]. 解剖学报, 2023, 54(6): 635-643.
[13]	魏纯纯王平林方兴麻献华章卫平谢志芳 . 小鼠棕色脂肪透射电子显微镜样品固定方法的改良 [J]. 解剖学报, 2023, 54(6): 738-743.
[14]	阎锟武筱林刘英娟葛科立任雷鸣李红云 . 脑蛋白水解物-Ⅰ对帕金森病小鼠肠道菌群的调节作用[J]. 解剖学报, 2023, 54(5): 497-504.
[15]	李梦一吴安婷许泽婷张庭李军伟周鹏崔怀瑞孙臣友 . 哺乳动物雷帕霉素靶蛋白复合物2/Akt信号通路对6-羟基多巴胺处理的SH-SY5Y细胞模型的作用及分子机制[J]. 解剖学报, 2023, 54(5): 521-530.