实验性重症肌无力大鼠模型的建立及巨噬细胞在发病机制中的作用探讨

The establishment of experimental autoimmune mysathenia gravis(EAMG) and study of the role of macrophage in the pathogenesis of EAMG

目的探讨早期重症肌无力发病机理中巨噬细胞的作用及巨噬细胞相关的细胞因子和趋化因子的变化。方法用原位杂交探索在EAMG动物模型靶器官肌肉组织中细胞因子和趋化因子mRNA的表达。采用原位杂交和免疫细胞化学染色双染技术,检测在Lewis大鼠EAMG早期,巨噬细胞浸润和程序性细胞凋亡。结果在EAMG早期观察在7和12天的EAMG大鼠肌肉组织中可见TNF-α、IL-1β和IL-6mRNA的表达细胞瞬时爆发,其峰值分别是(8.7±2.2)、(13.1±1.2)和(6.9±2.4)cells/mm2。第15天TNF-αmRNA的表达细胞数量再一次上升,IFN-γ、IL-4、IL-10、TGF-β和溶细胞素的mRNA表达细胞非常低。这些细胞主要在EAMG的早期观察到,范围是1～4cells/mm2。结论(1)EAMG早期大量巨噬细胞浸润,程序性细胞凋亡是浸润的巨噬细胞消除的主要方式;(2)EAMG早期于肌肉组织中细胞因子低水平表达;没有发现C-C趋化因子。

Objective：To explore the function of macrophage and the change of cytokine and chemokine related to macrophage in the early experimental autoimmune myasthenia gravis （EAMG）. Methods： In-situ hybridization was applied to explore cytokine and chemokine mRNA expression in sections of striated muscle. Combining in situ tailing and irmnanocytochemical staining was applied to detect the infiltrating macrophages and apoptotic macrophages in muscle tissue sections during early phase of EAMG in Lewis rats. Results： A transient burst of TNF-α, IL-1β and IL-6 mRNA expressing cells was detected during the early phase of EAMG, the peak values of which were （ 8. 7 ±2. 2）, （ 13.1 ± 1.2） and （6. 9 ±2. 4） cells/mm2. TNF-α mRNA expressing cells elevated in the 15th day gain. During the early phase of EAMG. !evels of IFN-γ, IL-4, IL-10, cytolysin and TGF-β mRNA expressing cells were low, the range of which was 1-4 cells/mm^2. Conclusion：Apoptosis is the major cause for elimination of infiltrating macrophages during the early phase of EAMG. During the early phase of EAMG cytokine mRNA expressing cells are low. C-C chemokine mRNA expressing cells are not detected.