应用基因芯片技术检测大鼠急性脊髓损伤早期阶段的基因表达谱  

Gene expression profile in acute spinal cord injury screened by cDNA microarray:experiment with rats

作　　者：董大明 姚猛 李克深[1] 王岩松 

机构地区：[1]哈尔滨医科大学分子流行病学教研室

出　　处：《中华医学杂志》2006年第35期2495-2501,共7页National Medical Journal of China

摘　　要：目的探讨急性脊髓损伤(SCI)后的特征性改变的分子机制。方法 SD 大鼠9只随机分为3组:SCI 8 h 组(3只)、72 h 组(3只)及对照组(3只),用改良的 Allen's 撞击法建立大鼠脊髓挫伤模型,应用含有13 200条大鼠基因的 cDNA 表达谱基因芯片对大鼠脊髓外伤性损伤后8 h 和72 h 的基因表达谱变化进行分析,并用 RT-PCR 方法对其中12条基因的表达水平变化进行验证。C/EBPδ基因的表达水平变化采用免疫组化和原位杂交方法进行验证。结果 SCI 8 h,52个基因的表达出现显著性变化,上调基因包括转录因子、早期快反应基因、氧化应激、补体、促炎症及抗炎症反应基因等;下调基因包括离子通道、突触及囊泡相关蛋白、神经丝蛋白、细胞骨架相关蛋白和髓鞘相关糖蛋白等。SCI 72 h,44个基因的表达出现显著性变化,包括生长因子、GTP 酶、癌基因及抑癌基因、泛素系统相关蛋白等,这些基因在神经元的生长/分化/存活、修复和再生等方面起重要作用。12条基因 RT-PCR 的验证结果与芯片的结果相符合。结论 SCI 早期多种基因表达出现显著性变化,C/EBPδ是一个在损伤脊髓组织中高表达的基因,很可能是一个治疗 SCI 介入靶点。Objective To investigate the characteristic changes of expression of the genes with specific functions in acute spinal cord injury （SCI）. Methods Nine SD rats were randomly divided into 3 equal groups： SCI 8 -hour group in which modified Allen＇s falling strike method was used to establish spinal cord contusion model, the spinal cords were taken out 8 hours later to undergo examination of the gene expression profde by using eDNA mieroarray including 13 200 gene, 12 genes were selected to undergo semiquantitative RT-PCR, and the up-regulation of the candidate gene C/EBPδ was verified by in situ hybridization and immunohistoehemistry. ; SC172 -hour group undergoing the same treatment, however, with the spinal cord taken out 72 hours later; and control group undergoing only sham operation with the spinal cord taken out immediately. Results In the SCI 8 hour group the expression of 52 genes differed in comparison with the control group, 30 genes, including those related to transcription factors, oxidative stress, complement, pro-inflammatory reaction, and anti-inflammatory reaction, were up-regulated and 22 genes related to ion channel, synaptie proteins, and eytoskeletal proteins, were down-regulated. In the SCI 72-hour group the expression of 44 genes with known functions related to growth/differentiation/survival, axonal guidance, neuron regeneration, signal transduetion, ubiquitin-proteasome system, and tumor suppressor differed, 26 genes were up-regulated and 18 down-regulated, in comparison with the control group. Semi-quantitative RT-PCR results of the 12 genes were consistent with those by the mieroarray examination. Conclusion Significant changes occur in the early stage of SCI. Expressed at a high level in SCI, C/EBPδ may be a therapeutic target of SCI.

关 键 词：脊髓损伤 基因表达 基因芯片 

分 类 号：R651.2[医药卫生—外科学]