【摘要】 目的建立进展型实验性自身免疫性脑脊髓炎(experimental autoimmune encephalomyelitis,EAE)模型,观察中枢神经系统(central nervous system,CNS)不同部位在病变不同阶段的组织病理学变化以及M1型、M2型小胶质细胞的动态变化。方法以C57BL/6为实验小鼠,建立EAE模型,于不同发病时期对CNS各部位进行切片;髓鞘染色(luxol fast blue,LFB)检测髓鞘脱失、苏木素-伊红(hematoxylin-eosin, HE)染色检测炎细胞浸润;免疫组织化学(immunohistochemistry,IHC)检测M1型、M2型小胶质细胞的动态变化;Real-Time PCR检测腰髓段中iNOS,TNF-α,Arg-1以及IL-4的mRNA水平。结果 LFB和HE染色结果显示EAE小鼠CNS各部位均于发病点出现髓鞘脱失和炎细胞浸润,且随疾病进展逐渐加重。IHC结果显示,M1型小胶质细胞浸润随疾病进展不断增加;M2型浸润先增加后减少。Real-Time PCR结果显示,iNOS,TNF-α的mRNA水平随疾病进展逐渐增加;Arg-1,IL-4的mRNA的表达水平为先增加后降低。结论在进展型EAE小鼠的病理变化中,CNS各部位髓鞘脱失程度和炎细胞浸润随疾病进展变得更加严重,且以腰髓段的病理变化最为严重。随着疾病进展,M1型小胶质细胞数量逐渐增加,M2型的浸润先增加后减少。更多还原

【Abstract】 Objective To observe the histopathological changes as well as the M1 and M2 microglia of the central nervous system(CNS) from the different parts at different stages in experimental autoimmune encephalomyelitis(EAE) mice.Methods: C57 BL/6 mice were used to establish EAE model. The parts of CNS were collected respectively at different time point during disease progression. Hematoxylin-eosin(HE) staining and luxol fast blue(LFB) staining were used to detect inflammatory cell infiltration and the loss of myelin sheath. The immunohistochemistry analysis was used to test the infiltration of M1 and M2 microglia in the different parts of CNS. Real-time PCR was used to detect mRNA levels of iNOS, Arg-1, TNF-α and IL-4 in the lumbar spinal cord. Results: HE and LFB staining results showed that inflammatory cell infiltration and myelin loss emerged at the disease onset in the parts of CNS in EAE mice, and sustained to exacerbate the progression of this disease. The immunohistochemical results showed that the number of M1 microglia continued to increase, and the number of M2 microglia increased first and then decreased with disease progression. Real-Time PCR results showed that iNOS and TNF-α mRNA levels in the lumbar spinal cord continued to increase with the progression of the disease, the mRNA levels of Arg-1 and IL-4 increased first and then decreased.Conclusion: Inflammatory cell infiltration and myelin loss gradually aggravated in CNS of EAE mice during the disease progression. The number of M1 microglia was increasing, whereas the amount of M2 microglia increased first and then decreased.更多还原