t-PA对EAE病理性淋巴细胞与血脑屏障粘附的影响  

Effect of t-PA on Adhesion of Lymphocyte Blood-brain Barrier Endothelium in Experimental Autoimmune Encephalomyelitis Mice

作　　者：王菁华[1,2] 张建华[1,2] 于瑞洪[1,2] 穆莉莉[1,2] 刘玉梅[1,2] 刘传亮[1,2] 王广友[1,2] 

机构地区：[1]哈尔滨医科大学神经生物学教研室 [2]哈尔滨医科大学神经生物学省重点实验室,黑龙江哈尔滨150081

出　　处：《现代生物医学进展》2015年第27期5208-5211,5220,共5页Progress in Modern Biomedicine

基　　金：黑龙江省教育厅科学技术研究项目(12521208)

摘　　要：目的:研究组织型纤溶酶原激活剂(t-PA)对实验性自身免疫性脑脊髓炎(EAE)小鼠病理性淋巴细胞与血脑屏障粘附的影响。方法:用MOG35-55肽段免疫C57BL/6小鼠建立EAE动物模型,于发病高峰期取淋巴细胞用MOG35-55肽段进行刺激得到抗原特异性T淋巴细胞。通过尾静脉给予t-PA的方法对EAE小鼠进行干预,临床评分评价小鼠的发病情况。体外培养小鼠血脑屏障内皮细胞系b End.3,应用不同浓度的t-PA进行处理。用荧光标记MOG35-55特异性T细胞进行细胞粘附实验,用Transwell小室建立体外血脑屏障模型进行细胞迁移实验。用免疫荧光化学方法检测ICAM-1的表达情况。结果:t-PA处理可以使血脑屏障内皮细胞与T淋巴细胞粘附和迁移作用增强。在体外细胞培养模型中检测到t-PA诱导ICAM-1表达升高。经过t-PA处理的小鼠,其血管内皮表面ICAM-1的表达也有所上升。经t-PA处理的EAE小鼠发病高峰提前,症状加重。结论:t-PA处理可以使EAE病理性淋巴细胞与血脑屏障内皮的粘附性增加,浸润能力增强;t-PA所引起的粘附性增加可能与b End.3表面ICAM-1表达升高有关。Objective： To investigate the effects of tissue-type plasminogen activator（t-PA） on T cell-brain microvascular endothelial cell adhesionin experimental autoimmune encephalomyelitis（EAE） mice. Methods： EAE was induced in C57BL/6 mice with immunization of MOG35-55 peptides. T cells were obtained from lymph nodes of EAE mice 15 d after immunization, and then stimulated with MOG35-55 peptides to obtain the MOG specific T cells. t-PA administration was performed via tail vein injections,animals were monitored for signs of disease and were scored. Mouse brain-derived endothelial cell line, b End3, was treated with various concentrations of t-PA. MOG specific T cells were labeled with Calcein AM to perform the adhesion assay. Transwell was used as the blood-brain barrier model to perform the cell Migration assay. Immunofluorescence was performed to detect the expression of ICAM-1.Results： In the presence of t-PA, enhanced adhesion and migration of T cells to b End.3 cells was observed. t-PA administration can induce ICAM-1 expression in mouse cerebral microvascular endothelial cells in vitro. Administration of t-PA in mice was also associated with an increase in ICAM-1 expression by brain endothelial cells. By using an EAE mouse model, t-PA treatment leads to the early onset of EAE and increased disease severity. Conclusion： t-PA administration can enhance adhesion of T cells to b End.3 cells which may associate with the increase in ICAM-1 expression on brain endothelial cells by t-PA.

关 键 词：组织型纤溶酶原激活剂 实验性自身免疫性脑脊髓炎 血脑屏障 粘附 

分 类 号：Q95-31[生物学—动物学] R744.51[医药卫生—神经病学与精神病学]