中文摘要：

目的比较WAR-5与盐酸法舒地尔（Fasudil）对实验性自身免疫性脑脊髓炎（EAE）的治疗效果,并探究其作用机制。方法采用小鼠髓鞘少突胶质细胞糖蛋白35-55多肽（MOG35-55）免疫雌性C57BL/6小鼠建立慢性EAE模型,并随机分为EAE对照组、WAR-5治疗组和Fasudil治疗组。于免疫后第3天开始,分别腹腔注射WAR-5、Fasudil及生理盐水,比较各组小鼠体质量变化和临床评分。于免疫后第28天处死各组动物,制作脊髓冰冻切片进行CD4＋T细胞、CD68＋巨噬细胞及核转录因子-κB（p-NF-κB/p65）染色,观察其在脊髓中的表达。提取脑组织的蛋白用Western blot法检测Rho相关卷曲螺旋蛋白激酶（ROCK）Ⅱ和p-NF-κB/p65蛋白表达,并采用Griess法检测脾细胞培养上清液的一氧化氮（NO）释放水平。结果与EAE对照组（108.70±20.26、399.00±25.94）比较,WAR-5治疗组（13.67±4.041、52.67±13.43）及Fasudil治疗组（15.33±5.033、31.00±13.45）脊髓内CD4＋T细胞和CD68＋巨噬细胞数目均减少（均P〈0.01）,WAR-5与Fasudil两治疗组相比,差异无统计学意义。与EAE对照组（1.9500±0.3976、0.5885±0.05982）比较,WAR-5组（0.6137±0.1778、0.0096±0.0015）及Fasudil组（1.1960±0.2225、0.3574±0.1496）脑内ROCKⅡ、p-NF-κB/p65表达均降低,WAR-5治疗组脑内ROCKⅡ、p-NF-κB/p65表达亦均低于Fasudil治疗组（均P〈0.05）。WAR-5和Fasudil治疗组上清液NO含量分别为（16.40±0.5247）μmol/L、（17.69±0.2833）μmol/L,均低于EAE对照组〔（20.07±0.6116）μmol/L;均P〈0.001〕,且WAR-5治疗组低于Fasudil治疗组（P〈0.01）。与Fasudil治疗组比较,800μg及1600μg药物注射后,WAR-5治疗组小鼠的存活状态明显较好;800μg药物注射后,WAR-5治疗组小鼠的脚趾无明显血管扩张反应。结论 WAR-5对EAE的治疗效果与Fasudil相近,且其治疗安全性较好,扩张血管作用较小,其作用机制可能与WAR-5抑制小鼠ROCKⅡ的表达及抑制炎性反应相关?

英文摘要：

Objective To compare the therapeutic effect of WAR-5and Fasudil in experimental autoimmune encephalomyelitis（EAE）,and explore the possible mechanisms.Methods Female C57BL/6 mice were immunized with myelin oligodendrocyte glycoprotein peptides 35-55（MOG35-55）to establish chronic EAE model.Mice were randomly divided into three groups：the WAR-5treatment group,the Fasudil treatment group and the EAE control group.WAR-5,Fasudil or normal saline were intraperitoneally injected from day 3postimmunization（p.i）to day 27 p.i.The body mass changes and clinical scores of mice were compared among the three groups.On day 28,p.i.animals were sacrificed and spinal cords frozen sections were prepared.Immunohistochemistry staining method was adopted in CD4＋T cell,CD68＋macrophage and p-NF-κB/p65.Their expression in the spinal cord was observed.Protein from brains were extracted and ROCKⅡand p-NF-κB/p65 were detected by Western blot.Nitric oxide（NO）release levels in supernatants of spleen cell cultures were detected by Griess method.Results Compared with the EAE control group（108.70±20.26、399.00±25.94）,CD4＋T cell and CD68＋macrophage number in spinal cord decreased in the WAR-5treatment group（13.67±4.041、52.67± 13.43）and the Fasudil treatment group（15.33 ± 5.033、31.00 ± 13.45,P 0.01,respectively）.There was no significant difference between the WAR-5treatment group and the Fasudil treatment group.Compared with the EAE control group（1.9500±0.3976、0.5885±0.05982）,expression of ROCK Ⅱ、p-NF-κB/p65 in brains decreased in the WAR-5treatment group（0.6137±0.1778、0.0096±0.0015）and the Fasudil treatment group（1.1960±0.2225、0.3574±0.1496）.Expression of ROCK Ⅱ、p-NF-κB/p65 in brains of the WAR-5 treatment group was lower than that of the Fasudil treatment group（P 0.05,respectively）.Supernatant NO contents in the WAR-5treatment group 〔（16.40±0.5247）μmol/L〕and the Fasudil treatment group 〔（17.69±0.2833）μmol/L〕were lower than that of the EA