Src抑制的蛋白激酶C底物在实验性自身免疫性脑脊髓炎大鼠脊髓中的表达及其意义

Expression and significance of Src-suppressed C kinase substrate in rat models of experimental autoimmune encephalomyelitis

目的研究Src抑制的蛋白激酶C底物(Src-suppressed C kinase substrate,SSeCKS)在实验性自身免疫性脑脊髓炎(experimental autoimmune encephalomyelitis,EAE)进程中的表达情况,探讨其在EAE病理过程中可能的功能及意义,为多发性硬化(multiple sclerosis,MS)和EAE提供新的治疗靶点。方法构建EAE大鼠模型,进行临床打分评估;用HE染色检测EAE大鼠脊髓组织的炎性浸润情况;用western blot检测SSeCKS的表达变化;用免疫组织化学观察SSeCKS在脊髓组织中的分布。结果髓磷脂碱性蛋白(myelin basic protein,MBP)诱导单相的EAE过程:Lewis大鼠经MBP免疫后7～10d开始有临床症状,13～16d达到发病高峰,之后自发地恢复,至30d左右恢复到病前水平;EAE病程高峰期,大鼠脊髓炎性浸润情况显著;SSeCKS蛋白表达在EAE起始E1期显著增加,E3期达到高峰,之后开始下降,恢复期基本恢复到正常水平。结论本实验成功构建了Lewis大鼠的EAE模型;EAE病理过程中,SSeCKS蛋白表达水平发生变化,提示SSeCKS可能参与了EAE过程。

Objective To observe the expression patterns of Src-suppressed C kinase substrate（SSeCKS） in rat models of experimental autoimmune encephalomyelitis（EAE）,and investigate the possible functions of SSeCKS in the pathogenesis to find new therapeutic strategies against EAE and multiple sclerosis（MS）.Methods The model was prepared in Lewis rat induced by myelin basic protein（MBP）.The rats were sacrificed at different phase after immunization.Inflammatory cell infiltration was observed by H-E staining.Western blot was used to detect the changes of SSeCKS expression during EAE and the distribution of SSeCKS in spinal cord of EAE rats was investigated by immunohistochemisty staining.Results Monophasic EAE process was induced by MBP in Lewis rats.EAE onset in rats occurred during the days 7 to 10 after immunization,peaked at the days 13 to 16,and exhibited spontaneous remission until the day 30.The rats at peak of EAE presented significant inflammatory cell infiltration,but restored their pathological phenotype in the recovery phase.The expression of SSeCKS exhibited low level in the spinal cord of normal rats and CFA controls,but markedly increased from E1 stage to remission phase with a peak at the E3 stage after EAE induction.SSeCKS was found to locate in neurons and glias.Conclusion The rat model of EAE was successfully established.SSeCKS expression changed during EAE course,which suggested SSeCKS may involve in the development of EAE.