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目的研究EAE大鼠胸腺细胞凋亡的变化规律,为探讨胸腺在EAE发病中的作用提供理论依据。方法应用光镜、电镜观察正常组、EAE发病第3天组、恢复期组大鼠胸腺细胞变化情况,应用caspase-3免疫组化染色及流式细胞术观察比较胸腺细胞凋亡情况。结果电镜、光镜观察显示正常组和恢复期组胸腺结构基本一致,皮质髓质分界清楚,淋巴细胞密集,核呈蓝紫色,发病第3天组胸腺明显萎缩,皮质变薄,淋巴细胞数减少,并可见凋亡淋巴细胞;caspase-3免疫组化染色显示发病第3天组胸腺皮质凋亡淋巴细胞数目明显增多;流式细胞术显示发病第3天组大鼠胸腺细胞凋亡率显著升高(P<0.05),恢复期组凋亡率与正常组相比无统计学意义(P>0.05)。结论 EAE大鼠发病后第3天胸腺细胞明显减少并发生凋亡,随病情恢复淋巴细胞增加、凋亡细胞减少,表明胸腺具有自我恢复能力,其变化与EAE形成有关。
Objective To study the changes of thymocyte apoptosis in EAE rats and to provide a theoretical basis for exploring the role of thymus in the pathogenesis of EAE. Methods The changes of thymocytes in normal group, EAE on day 3 and convalescent group were observed by light microscope and electron microscope. The apoptosis of thymocytes was observed by immunohistochemistry and flow cytometry. Results Electron microscopy and light microscopy showed that the thymus structure was basically the same in normal group and convalescent group. The demarcation of ciliated medulla was clear, the lymphocytes were dense, the nucleus was blue-purple, the thymus was obviously atrophied on the 3rd day, the cortex became thinner and the number of lymphocytes decreased , And apoptotic lymphocytes were observed. The number of apoptotic lymphocytes in the thymus cortex increased obviously on the 3rd day after the onset of the caspase-3 immunohistochemical staining. Flow cytometry showed that the thymocyte apoptosis rate was significantly increased on the 3rd day (P <0.05). There was no significant difference between the recovery group and the normal group (P> 0.05). Conclusions The thymus cells in EAE rats were significantly reduced and apoptotic on the 3rd day after the onset of EAE. The number of apoptotic cells decreased with the increase of lymphocytes recovered, indicating that the thymus has the ability of self-recovery. The changes of EAE are related to the formation of EAE.