阿米洛利对实验性自身免疫性脑脊髓炎小鼠中枢神经系统中细胞因子表达的影响

目的：观察阿米洛利对实验性自身免疫性脑脊髓炎（EAE）小鼠脑组织中白介素17（IL-17）、白介素10（IL-10）和干扰素-γ（IFN-γ）的影响。方法：C57BL/6小鼠27只随机分成3组：EAE-生理盐水组、EAE-阿米洛利组和空白对照组。用髓鞘少突胶质细胞糖蛋白和完全弗氏佐剂混合抗原乳剂诱导小鼠EAE动物模型。EAE-阿米洛利组免疫当日开始给予阿米洛利10 mg.kg-1干预,连续14 d;EAE-生理盐水组给予相同剂量的生理盐水;空白对照组小鼠不做任何处理。每天评估EAE小鼠神经功能变化;采用苏木精-伊红染色观察病理学改变;ELISA法检测发病高峰期（免疫后第19天）脑组织中促炎因子IL-17和IFN-γ、抑炎因子IL-10改变。结果：与EAE-生理盐水组相比,EAE-阿米洛利组①日均神经功能评分降低（P〈0.05）,最大临床症状评分明显降低（P〈0.01）;而两组间发病率无明显差异。②苏木精-伊红染色：中枢神经系统炎症细胞浸润减少（P〈0.01）。③IL-17和IFN-γ表达降低（P〈0.01）,IL-10表达增加（P〈0.05）。结论：阿米洛利能够减缓小鼠EAE病情,其机制可能与下调中枢神经系统IL-17和IFN-γ的表达,上调IL-10的表达有关。

Intervention Effects of Amiloride on Cytokines in the Central Nervous System of Mice with Experimental Autoimmune Encephalomyelitis

Aim： To investigate the effects of amiloride on experimental autoimmune encephalomyelitis（EAE） mice and the changes of the levels of IL-17,IFN-g and IL-10 in central nervous system（CNS）.Methods： C57BL/6 mice were randomly divided into three groups： vehicle group,EAE-saline group and EAE-amiloride treated group.EAE was induced with myelin oligodendrocyte glycoprotein（MOG 35-55）.Mice in EAE-amiloride treated group were administrated with amiloride for 14 days at the dose of 10 mg·g-1 after immunization,and mice in EAE-saline group were treated with saline as the same dose as amiloride.The behavioral changes and the daily clinical scores were recorded.The damage and the infiltration of inflammatory cells in CNS were evaluated with HE stain.Expression levels of IL-17,IFN-g and IL-10 in CNS were quantified by ELISA.Results： Compared with EAE-saline group,the severity of paralysis（P0.01） and the infiltration of inflammatory cells（P0.01） in CNS were improved significantly in EAE-amiloride treated mice.ELISA analysis showed that the expressions of IL-17 and IFN-g（P0.05） decreased significantly in the CNS in EAE-amiloride treated mice compared with EAE-saline group,and the expressions of IL-10 increased in the CNS in EAE-amiloride treated mice.Conclusion： Amiloride had protective effects on EAE,which may be due to the down regulations of IL-17 and IFN-g,shifting the balance of pro-inflammatory and anti-inflammatory cytokines.