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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 effect of tissue plasminogen activator (t-PA) on the adhesion of pathological lymphocytes and blood-brain barrier in experimental autoimmune encephalomyelitis (EAE) mice. Methods: Animal models of EAE were established by immunizing C57BL / 6 mice with MOG35-55 peptide. The lymphocytes were stimulated with MOG35-55 peptide to obtain antigen-specific T lymphocytes. EAE mice were intervened by t-PA tail vein injection, and clinical scores were used to evaluate the incidence of mice. The mouse BBB endothelial cell line b End.3 was cultured in vitro and treated with different concentrations of t-PA. Cell adhesion experiments were performed with fluorescently labeled MOG35-55-specific T cells. Transwell chambers were used to establish an in vitro model of blood-brain barrier for cell migration. ICAM-1 expression was detected by immunofluorescence staining. Results: t-PA treatment could enhance the adhesion and migration of blood-brain barrier endothelial cells and T lymphocytes. T-PA was found to induce elevated ICAM-1 expression in in vitro cell culture models. After t-PA treatment of mice, the expression of ICAM-1 on the vascular endothelial surface also increased. EAE mice treated with t-PA peaked earlier and the symptoms worsened. Conclusion: The t-PA treatment can increase the adhesion of EAE pathological lymphocytes to the blood-brain barrier endothelium and enhance the infiltration capacity. The increased adhesion induced by t-PA may be related to the increase of ICAM-1 expression on the surface of b End.3 High related.