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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。
Objective: To observe the effect of amiloride on the expression of interleukin 17 (IL-17), interleukin 10 (IL-10) and interferon-γ (IFN-γ) in the brain of experimental autoimmune encephalomyelitis influences. Methods: Twenty-seven C57BL / 6 mice were randomly divided into 3 groups: EAE-saline group, EAE-amiloride group and blank control group. Mouse EAE animal model was induced by mixing antigen emulsion with myelin oligodendrocyte glycoprotein and complete Freund’s adjuvant. In the EAE-amiloride group, amiloride 10 mg.kg-1 was administrated on the day of immunization for 14 consecutive days. The rats in the EAE-saline group were given the same dose of saline. The blank control mice were given no treatment. The neurological changes of EAE mice were evaluated every day. Pathological changes were observed by hematoxylin-eosin staining. The levels of proinflammatory cytokines IL-17 and IFN-γ in brain tissue were detected by ELISA at the peak of the disease (day 19 after immunization) The inflammatory cytokine IL-10 changes. Results: Compared with EAE-saline group, the mean daily neurological score of EAE-amiloride group was significantly lower (P <0.05) and the maximum score of clinical symptoms was significantly lower (P <0.01); while the morbidity Significant differences. ② hematoxylin-eosin staining: central nervous system inflammatory cell infiltration decreased (P <0.01). (3) The expression of IL-17 and IFN-γ decreased (P <0.01), and the expression of IL-10 increased (P <0.05). CONCLUSION: Amiloride can slow the progression of EAE in mice. The mechanism may be related to down-regulating the expression of IL-17 and IFN-γ in the central nervous system and up-regulating the expression of IL-10.