芬戈莫德对实验性自身免疫性脑脊髓炎小鼠脑组织基质金属蛋白酶-9表达影响的研究

The Effect of FTY720 on the Expression of MMP-9 in the Brain Tissue of Experimental Autoimmune Encephalomyelitis Mice

目的探讨芬戈莫德对实验性自身免疫性脑脊髓炎(EAE)小鼠脑组织基质金属蛋白酶-9(MMP-9)mRNA及蛋白表达的影响。方法 48只雌性C57BL/6小鼠随机分为完全弗氏佐剂(CFA)组、EAE组、芬戈莫德组。运用髓鞘少突胶质细胞糖蛋白35-55构建EAE小鼠模型,小鼠神经功能缺损评分达到1分或免疫第14天,芬戈莫德组腹腔注射芬戈莫德10 mg·kg-1,相应的CFA组、EAE组给予生理盐水。观察小鼠行为学变化,中枢炎症和脱髓鞘情况,脑组织中的MMP-9 mRNA及蛋白表达。结果芬戈莫德组小鼠临床症状减轻,体重下降减少,潜伏期延长,发病率降低,炎性病灶数减少,脱髓鞘程度减轻。芬戈莫德组MMP-9 m RNA和蛋白表达水平较EAE组降低。结论芬戈莫德能抑制EAE小鼠脑组织中MMP-9 mRNA及蛋白表达水平。

Aim To investigate the effect of fingolimod（FTY720） on the expression of matrix metalloproteinase-9（MMP-9） in the brain tissue of experimental autoimmune encephalomyelitis（EAE）. Methods Forty-eight C57BL/6 mice were randomly divided into three groups equally： EAE group（made by using myelin oligodendrocyte glycoprotein（MOG35-55）, treated with normal saline）, CFA group（made by using normal saline instead of MOG35-55, treated with normal saline）, FTY720 group（made as EAE group, intraperitoneal injected FTY720（10 mg·kg-1）. Mice were examined daily for clinical signs and scored accordingto the signs and symptoms. Hematoxylin-eosin staining and Luxol Fast Blue myelin staining were performed on the sections of brain. The number of inflammation focus in the brain tissue was counted under the optical microscope. The expressions of MMP-9 in brain tissue were detected by immunohistochemistry technique, while mR NA expression of MMP-9 in mice were detected by real-time PCR technology. Results Comparing to EAE group, neurological deficit scores, weight 1oss, eclipse period and morbidity decreased in FTY720 group, as well as the amount of the inflammation focus and demyelination lesions. The expression level of MMP-9 and MMP-9 mR NA in FTY720 group were lower than that of EAE group. Conclusion FTY720 inhibit the expression level of MMP-9 protein and mR NA in the brain tissue of EAE mice.