摘要
目的:观察正常培养、诱导凋亡以及益气化瘀中药干预后的大鼠椎间盘软骨细胞基因表达水平的变 化,研究益气化瘀中药对椎间盘凋亡软骨细胞基因表达谱的调节作用。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、诱导凋亡组和益气化瘀中药组的大鼠颈椎间盘软骨细胞 中抽提mRNA,经反转录后分别用Cy3、Cy5荧光标记,获得椎间盘软骨细胞cDNA的探针,再与 BiostarR 40s的基因表达谱芯片杂交,ScanArray4000激光扫描仪扫描,GenePixPro3.0图像处理软件进 行图像分析、标准化处理和ratio值分析。结果:诱导凋亡组与正常组比较,共筛选出30条表达有差异的已 知基因,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);中药组与凋亡组比 较,共筛选出97条表达有差异的已知基因,中药上调44条基因(ratio>2),下调53条基因(ratio<0.5),涉 及到丝分裂素激活蛋白激酶(mitogen activatedproteinkinase,MAPK)和信号转导和激活因子(signal transducerandactivatoroftranscription,STAT)。结论:益气化瘀中药对椎间盘凋亡软骨细胞的调控涉及 多个方面,其对MAPK和JAK STAT通路的影响可提示益气化瘀中药对信号转导通路的调控作用。这从 一个侧面探索了中医气血理论与信号转导的关系。
Objective： To study the effects of Yiqi Huayu Recipe on the gene expression pattern in normal and apoptotic chondrocytes in the cervical intervertebral disc of rats. Methods： The intervertebral disc endplates of rats were digested enzymatically. The apoptosis of the chondrocytes was induced by anti-Fas antibody. BiostarR-40s microarray chips were used to investigate the gene expression pattern in chondrocytes in the normal control, apoptosis and Yiqi Huayu Recipe groups. The results were scanned with Scan Array 4000 and analyzed with GenePix Pro 3.0, and then subjected to standardization and ratio analysis. Results： In the apoptosis group, 30 kinds of genes expressed in cervical intervertebral disc chondrocytes were screened out,in which 10 were up-regulated （ratio〉2） and 20 down-regulated （ratio〈0.5） as compared with the normal control group. On the other hand, in the Yiqi Huayu Recipe group, 97 kinds of genes expressed in cervical intervertebral disc chondrocytes were screened out, in which 44 were up-regulated （ratio〉2） and 53 down-regulated （ratio〈0.5） as compared with the apoptosis group. Conclusion： There are some signal transduction pathways that control the apoptosis of the intervertebral disc chondrocytes. Yiqi Huayu Recipe regulates the gene express of the apoptotic chondrocytes in intervertebral disc. The study gives a further understanding to the mechanism of the cervical spondylosis and enriches the theories of qi and blood of traditional Chinese medicine.
出处
《中西医结合学报》
CAS
2005年第2期111-114,共4页
Journal of Chinese Integrative Medicine
基金
国家自然科学基金重点资助项目(No.30171170
30330700)
上海市"启明星"基金资助项目(No.01QB14039)
关键词
益气化瘀方
椎间盘
软骨细胞
凋亡
基因芯片
信号转导
Yiqi Huayu Recipe
intervertebral disc
chondrocytes
apoptosis
gene chip
signal transduction
