The roles of cytokines and neurotrophins in the regenerative process of nervous system

在神经系统的不同区域研究了细胞因子和神经营养/神经保护因子在损伤/再生过程中的作用。大鼠制作三种损伤模型：(1)坐骨神经慢性挤压伤(CCI)或轴索切断；(2)脊髓挤压伤；(3)局灶性脑缺血/再灌注模型。研究IEGs、细胞因子和神经营养因子的蛋白质和mRNAs的时空表达模式，并确定表达这些分子的细胞类型。为了探讨IL-6家族细胞因子在神经系统中的作用，研究了IL-6家族细胞因子gp130、LIFRβ、IL-6R、OSMR以及JAK-STAT通路的信号转导分子在大鼠和小鼠脑内的表达。我们发现这些受体成分在小脑浦肯野细胞中表达。激光诱导因子过表达诱导浦肯野细胞…树突状细胞明显萎缩更多。在体外培养的颈上神经节模型中，加入血清可抑制树-树突触的重组，提示血清衍生因子参与了神经元树突形态的调节。在肌肉、角膜和胃等周围组织的实验表明，细胞因子在损伤后IEGs表达触发的再生过程中发挥着重要作用。在c-fos基因敲除小鼠和c-fos和c-jun反义寡核苷酸处理的培养角膜中，角膜上皮细胞没有启动迁移。我们发现AP-1也参与了血管平滑肌细胞对BDNF和NT-3基因表达的调节。AP-1对NT-3基因的表达有负调控作用。构建了在CAG启动子控制下插入绿色荧光蛋白报告基因的E1a和E1B缺失腺病毒(ADV)，并用于体内实验。将这种AdV-EGFP注射到坐骨神经或皮肤中会导致DRG神经元的标记。较少

The involvement of cytokines and neurotrophic/neuroprotective factors in the process of injury/regeneration were investigated in various regions of the nervous system. Three types of injury models were prepared in rats ; (1)chronic constriction injury (CCI) or axotomy of the sciatic nerve, (2) crush injury of the spinal cord and (3) focal cerebral ischemia/reperfusion model. The spacial and temporal expression patterns of proteins and mRNAs of IEGs, cytokines and neurotrophins were investigated and cell types expressing thse molecules were determined. Expression of receptor components of IL-6 family cytokines, such as gp130, LIFRβ, IL-6R, OSMR, and signal transduction molecules of JAK-STAT pathway were investigated in the brain of rats and mice, in order to explore the roles of cytokines of this family in the nervous system. We found that these receptor components were expressed in cerebellar Purkinje cells. Overexpression of LIF in mice induced marked dendritic atrophy of Purkinje cel … More ls. Reorganization of dendro-dendritic synapses in the ex vivo model of superior cervical ganglion was inhibited by addition of serum in the medium, indicating that serum-derived factors are involved in the regulation of dendritic morphology of neurons. Experiments in the peripheral tissues, such as muscles, cornea and stomach, indicated that cytokines play important roles in the regeneration process, which is triggered by IEGs expression after injury. Migration of corneal epithelial cells was not initiated in c-fos knockout mice and in cultured cornea treated with antisense ODNs for c-fos and c-jun mRNAs. We showed that AP-1 is also involved in the regulation of BDNF and NT-3 gene expression using vascular smooth muscle cells. AP-1 was shown to negatively control the expression of NT-3 gene. E1A-and E1B-deleted adenovirus (AdV) containing EGFP reported gene insert under the control of CAG promoter was constructed and used in the in vivo experiment. Injection of this AdV-EGFP into the sciatic nerve or in the skin resulted in labeling of DRG neurons. Less