摘要
目的分析正常和诱导凋亡的大鼠颈椎间盘软骨细胞基因表达水平的变化。方法建立大鼠颈椎间盘软骨细胞培养体系,使用抗Fas抗体诱导凋亡,分别从正常组和诱导凋亡组的大鼠颈椎间盘软骨细胞中抽提mRNA,经反转录后用Cy3、Cy5荧光标记,获得椎间盘软骨细胞cDNA的探针,再与B iostarR-40 s的基因表达谱芯片杂交,ScanArray 4 000激光扫描仪扫描,GeneP ix Pro 3.0图像处理软件进行图像分析、标准化处理、Ratio值分析、聚类分析。结果凋亡细胞与正常细胞比较,共筛选出明显变化的基因231条。在30条表达有差异的已知基因中,20条基因表达量明显下降(Ratio<0.5),10条基因表达量明显上升(Ratio>2)。变化的基因包括蛋白酶类,生长因子类,信号转导通路类等方面。结论初步筛选出大鼠颈椎间盘软骨细胞的凋亡相关基因,为研究颈椎病在基因水平上的病理变化提供了基础。
Objective To compare the gene expressions of normal chondrocyte with apoptosis chondrocyte in rats cervical intervertebral disc. Methods The intervertebral disc endplate of rats were digested enzymatically. Apoptosis of cultured chondrocytes were induced by anti-Fas antibody. Biostar R-40s microarray chips were used to investigate the gene expression pattern in normal and apoptosy chondrocyte. The results were scanned and analyzed by ScanArray400 and GenePix3.0, and then subjected to standardization, ratio and clustering analysis. Results After comparison, 30 known genes were screened out, 10 of which unregulated （ ratio 〉 2 ） and 20 downregulated （ ratio 〈 0.5 ）. Conclusion There are differences between normal and apoptosis chondrocyte in rats cervical intervertebral disc in the gene expression patterns, the study providing information for clinical diagnosis and treatment of cervical intervertebral disease.
出处
《上海中医药大学学报》
CAS
2005年第4期45-47,共3页
Academic Journal of Shanghai University of Traditional Chinese Medicine
基金
国家自然科学基金资助项目(30171170)
上海市"启明星"基金资助项目(01QB14039)
