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

细胞因子和神经营养/神经保护因子在神经系统不同区域损伤/再生过程中的参与情况进行了研究。制备大鼠三种损伤模型；(1)坐骨神经慢性收缩损伤（CCI）或闭锁；(2)脊髓挤压损伤；(3)局灶性脑缺血再灌注模型。研究了脑电信号、细胞因子和神经营养因子的蛋白和mrna的时空表达模式，并确定了表达这些分子的细胞类型。通过研究IL-6家族细胞因子受体组分gp130、LIFRβ、IL-6R、OSMR及JAK-STAT通路信号转导分子在大鼠和小鼠脑内的表达，探讨IL-6家族细胞因子在神经系统中的作用。我们发现这些受体成分在小脑浦肯野细胞中表达。小鼠过表达LIF可诱导浦肯野细胞明显的树突萎缩。培养基中添加血清可抑制颈上神经节离体模型树突-树突突触的重组，提示血清源性因子参与了神经元树突形态的调控。在肌肉、角膜和胃等外周组织中进行的实验表明，细胞因子在损伤后的再生过程中发挥重要作用，并由egg表达触发。在c-fos敲除小鼠和c-fos和c-jun mrna的反义odn处理的培养角膜中，角膜上皮细胞没有开始迁移。我们发现AP-1也参与血管平滑肌细胞对BDNF和NT-3基因表达的调节。AP-1对NT-3基因的表达有负向控制作用。构建了CAG启动子控制下含有EGFP报道基因插入的e1a和e1b缺失腺病毒（AdV），并将其用于体内实验。将这种AdV-EGFP注射到坐骨神经或皮肤中导致DRG神经元的标记。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