【摘要】 目的利用基因芯片技术,比较风湿性二尖瓣狭窄病人和健康人外周血细胞因子基因表达的差异,从整体上认识风湿性二尖瓣狭窄的心脏瓣膜病的细胞因子表达状况。方法单纯性二尖瓣狭窄的病人3人,以健康人5人作为对照,应用Trizol法分别提取他们的外周血总RNA,逆转录合成cDNA,用Cy5、Cy3分别标记病人、对照组的cDNA,混合后在细胞因子表达谱芯片上杂交。结果经对原始数据的标准化和SAM芯片数据软件的应用,获得CXCR4,RGS2和IRAK13种显著性差异表达的细胞因子基因,均为表达上调的细胞因子基因,并对各自的作用进行归纳、分析。结论获得风湿性二尖瓣狭窄外周血细胞因子的基因表达图谱,并证实了风湿性二尖瓣狭窄的心脏瓣膜病是炎症反应和抗炎反应相互作用的结果,为进一步临床研究该病与细胞因子的关系提供一定的方向。更多还原

【Abstract】 Objective To compare the gene expression profile between peripheral blood of patients with isolated rheumatic mitral stenosis and that of healthy control,and to get a relative whole picture of the relationship between rheumatic mitral stenosis and cytokines,we used cytokine gene expression chip to study gene expression profile of cytokines in peripheral blood of patients with isolated rheumatic mitral stenosis. Methods Total RNAs were extracted from peripheral blood of three patients with mitral stenosis and from 5 healthy volunteers’ control group by Trizol methord. Then the total RNA was the template for the reverse transcription reaction that created the cDNA copy and directly incorporated a fluorescent nucleotide Cy3-dUTP for the control or Cy5-dUTP for the three patients. The two kinds of cDNA were mixed and hybridized on cytokine gene chip which contains 553 cDNA clones. Results Of the 553 genes monitored,three genes,including CXCR4,RGS2 and IRAK1,showed differential expression between patients with rheumatic mitral stenosis and healthy control,through data analysis by a sires of chip data softwares. Conclusions Our findings show that some uncommon cytokine genes significantly express in peripheral blood of patients with rheumatic mitral stenosis. These uncommon cytokines may play a role in rheumatic mitral stenosis,especially in non-rheumatic activity phase. It also proves that the valvular lesion of rheumatic mitral stenosis is the result of interaction between inflammatory response and anti-inflammatory response,and provides a way to deeply research the association between rheumatic mitral stenosis and cytokines expression.更多还原