益气化瘀方调控颈椎间盘细胞基因表达谱的研究

Effects of Yiqi Study on the Gene Expression Profile of Cervical Intervertebral Disc Requlated by Yiqi Huayu Fang in Rats

目的 :分析正常、退变SD大鼠颈椎间盘基因表达水平的变化 ,研究益气化瘀方对大鼠退变颈椎间盘基因表达谱的调节作用。方法 :建立动、静力失衡性大鼠颈椎间盘退变模型 ,分别从正常、退变和益气化瘀方组大鼠颈椎间盘中抽提mRNA ,经反转录后分别用Cy3、Cy5荧光标记 ,获得动物椎间盘cDNA的探针 ,再与 5 12点cDNA基因表达谱芯片杂交 ,激光扫描仪扫描并进行图像分析、标准化处理、ratio值分析、聚类分析。结果 :模型组与对照组比较 ,共筛选出 18条表达有差异的基因 ,11条基因表达量明显下降 (ratio <0 4 ) ,7条基因表达量明显上升 (ratio >2 5 )。益气化瘀方与模型组比较 ,共筛选出 2 2条表达有差异的基因 ,益气化瘀方上调 18条基因 (ratio >2 5 ) ,下调 4条基因 (ratio <0 4 )。其中在上述检测中表达最明显的是信号传导类基因 ,益气化瘀方能够上调多数细胞信号传导类基因 (如PTK等 )。结论 :初步筛选出大鼠退变椎间盘中相关基因 ,与已报道基因在颈椎间盘退变的表达模式类似 ,说明大鼠椎间盘退变受到各种基因的调控 ,初步阐明了颈椎病在基因水平上的发病机理。益气化瘀方参与了椎间盘细胞内相关基因的调控 ,参与椎间盘细胞内外的信号传导过程 ,从基因表达角度解释了中药治疗颈椎病的药效机理 ,丰富了中医?

Objective: To investigate the genes differential in normal and degeneration cervical intervertebral disc of rat. To study the effects of Yiqi Huayu Fang on the genes expression pattern in degenerated cervical intervertebral disc of rat. Methods: The model of rat cervical degeneration intervertebral disc caused by imbalance between the dynamic and static forces was established. There were control, model and Yiqi Huayu Fang groups, each group had 10 rats. Microarray chips containing 512 cDNAs were used to investigation on the gene expression pattern of cervical intervertebral disc of every groups, respectively. Results: There were 18 genes expressed in degenerated cervical intervertebral disc were screened out, comprising 11 up regulated and 7 down regulated, respectively. Yiqi Huayu Fang up regulates 18 genes and down regulate 4 genes in the degenerated discs. This genes express in cervical spondylosis was similar to other reports. Especially up regulate signal transduction gene showed that this prescription involved itself in the process of signal transduction pathways. Conclusions: The analysis of gene expression pattern of cervical intervertebral disc based on cDNA microarray can realize high throughput screening of genes associated with cervical intervertebral disc. Signal transudations pathways regulate the process of the degenerative intervertebral disc. Investigation on the gene expression pattern plays an important role in degenerative intervertebral disc pathogenesis. Yiqi Huayu Fang take part in gene express regulates of degenerated intervertebral disc. It can up regulation signal transduction genes (Focal adhesion kinase, et al) particular. These investigations can take a step forward expound cervical spondylosis mechanism of gene express and enrich theories of QI and Blood of T.C.M.