口服盐酸法舒地尔治疗小鼠EAE有效性初探  

Oral administration of fasudil hydrochloride suppresses development of experimental autoimmune encephalomyelitis in mice

作　　者：张辉[1,2] 张海飞[2,3] 李艳花[2] 刘春云[2] 尉杰忠[2] 丁智斌[1,2] 杨兴旺[2,5] 杨琬芳[5] 李俊莲[5] 冯前进[5] 丰玲[2] 肖保国[2,4] 马存根[1,2,5] 

机构地区：[1]山西医科大学第一临床医学院神经内科,山西太原030001 [2]山西大同大学脑科学研究所,山西大同037009 [3]大同市第五人民医院神经科,山西大同037009 [4]复旦大学华山医院神经病学研究所,上海200025 [5]山西中医学院第三中医院脑病科,卫生部国家临床重点专科,山西太原030024

出　　处：《中国病理生理杂志》2013年第11期2060-2065,共6页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81070957;No.81272163);山西中医学院“2011计划”培育计划资助项目(No.2011PY-1);2013年度国家国际科技合作专项项目(No.2013DFA30700);山西省青年自然科学基金资助项目(No.2012021034-2);山西大同大学青年科学研究项目(No.2010Q6)

摘　　要：目的:探讨盐酸法舒地尔口服对实验性自身免疫性脊髓炎(EAE)的治疗效果及机制。方法:采用髓鞘少突胶质细胞糖蛋白33-35肽(MOG35-55)免疫雌性C57BL/6小鼠建立慢性EAE模型,随机分为EAE模型组和法舒地尔治疗组。免疫后第3天,对照组予以灌胃生理盐水,治疗组予以法舒地尔,每天1次至免疫后第27天。比较2组小鼠体重变化和临床评分。第28天处死动物,脊髓冰冻切片进行髓鞘染色和HE染色,流式细胞术检测脾细胞M1型和M2型巨噬细胞的表型,ELISA法检测脾细胞培养上清液白细胞介素10(IL-10)、白细胞介素1β(IL-1β)和肿瘤坏死因子α(TNF-α)的释放。结果:法舒地尔口服推迟EAE发病时间,减缓EAE症状,减少体重下降,减少EAE髓鞘脱失和中枢神经系统炎症浸润,抑制M1型巨噬细胞CD16/32表达,增加M2型巨噬细胞CD206和IL-10表达,同时抑制炎症细胞因子IL-1β和TNF-α的释放。结论:法舒地尔口服具有良好的治疗效果,可明显减轻中枢神经系统的髓鞘脱失和炎症病灶。其作用机制可能使M1型巨噬细胞向M2型转化,从而抑制炎症细胞因子释放。AIM： To explore the therapeutic effect of fasudil hydrochloride by the oral route on the development of experimental autoimmune encephalomyelitis （EAE） in mice and its possible mechanism. METHODS ： The EAE model in female C57BL/6 mice was established by myelin oligodendrocyte glycoprotein 35-55 peptide（ MOG35-55 ） immunization and the immunized mice were randomly divided into saline control group and fasudil intervention group, in which saline and fasudil were administered by the oral route once every day from post-immunization （PI） day 3 to day 27. Clinical score and body weight were recorded every other day. On PI day 28, the spinal cords were obtained for HE and myelin staining. The splenocytes were isolated and the expression of CD16/32, CD206 and interleukin （IL） -10 was analyzed by flow cytometry. The levels of IL-1β and tumor necrosis factor α （TNF-ot） were detected by ELISA. RESULTS： Oral administration of fasudil delayed the onset of EAE, and attenuated the myelinoclasis of the model animals and the severity of EAE,accompanied by the phenotypic switch from M1 to M2 macrophages, the inhibition of the proinflammatory cytokine （IL-1β and TNF-α） production and the increase in IL-10release. CONCLUSION： Oral administration of fasudil exhibits thera- peutic effect on the development of EAE possibly through switching M1 macrophages to M2 phenotype and inhibiting inflam- matory responses in mice.

关 键 词：法舒地尔 口服 实验性自身免疫性脊髓炎 巨噬细胞 细胞因子类 

分 类 号：R320.54[医药卫生—人体解剖和组织胚胎学]