室间隔缺损患者外周血细胞因子基因表达状况的研究

Gene expression profile of cytokines in peripheral blood of patients with ventricular septal defect.

目的利用基因芯片技术,比较室间隔缺损病人和健康人外周血单个核细胞的细胞因子基因表达差异,从整体上认识室间隔缺损患者的细胞因子表达状况。方法通过检测102例室间隔缺损(VSD)患者外周血常规,并从中选择3例血小板平均体积不同数值的室间隔缺损的病人,另选5例健康人,应用总核糖核酸(RNA)的提取法(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、健康人的cDNA,混合后在细因子基因谱芯片上杂交,经对原始数据的标准化和SAM芯片数据软件的应用,获得表达有显著性差异的细胞因子基因。结果在102例VSD患者中,73例血小板平均体积低于正常值,占71.57%,3例血小板平均体积不同数值的患者均表现为27种细胞因子基因表达下调,一种细胞因子基因表达上调。其中β胸腺素-4和核糖体蛋白S5基因在三个样本中均表达下调,我们对各自基因的作用进行分析,了解可能的分子机制。结论分析室间隔缺损病人外周血细胞因子的基因表达图谱,β胸腺素-4和核糖体蛋白S5基因可能在心室发育中共同起重要的作用。

Objective By using gene chip technique to compare the gene expression profile between peripheral blood of patients with isolated ventricular septal defect and healthy controls, we got a relative whole view of the relationship between ventricular septal defect and cytokines. Methods 102 patients with VSD were tested for peripheral blood analysis. Three VSD patients with different mean platelet volume（MPV） values were selected for study. Total amount of RNA was extracted from peripheral blood of three VSD patients and five healthy volunteers by Trizol method, then the RNA was synthesized to cDNA and directly incorporated with fluorescent nucleotide Cy3 - dUTP for control group or Cy5 - dUTP for VSD patients. Two kinds of cDNA were mixed and hybridized on cytokine gene chip. After normalization, differential gene expression was assessed by significance analysis of microarrays （SAM）. Results Among 102 VSD patients, the mean platelet volume of 73 （ 71.57% ） patients was below the average level. The gene expression showed that 27 kinds of cytokine genes expressed in peripheral blood of three isolated VSD patients were down - regulated including Thymosin - beta - 4 and Ribosomal protein $5. Functions of these genes were analyzed and possible mechanism was found out by us. Conclusion Above - mentioned findings demonstrated that Thymosin - beta -4 and Ribosomal protein S5 may play an important role in development of ventricular septum.