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目的研究全基因组甲基化对心力衰竭患者心肌基因表达的影响。方法取心脏移植中供体左心室心肌标本8例作为正常对照组,行心脏移植的心力衰竭患者自身左心室心肌标本14例作为实验组,提取基因组基因通过甲基化DNA免疫共沉淀结合甲基化芯片技术(methylated DNA immunoprecipitation-chip,MeDIP-chip)及基因表达谱芯片进行高通量的快速筛选。然后,得到MeDIP-chip初筛后的2个甲基化异常基因abca4和cd200进行亚硫酸氢钠测序PCR。结果疾病组abca4基因启动子区甲基化率为85.5%,正常组基因启动子区甲基化率为91.2%;疾病组基因启动子区甲基化率比正常组基因启动子区甲基化率低5.7%,心衰组cd200基因启动子区甲基化率为50.5%,正常组基因启动子区甲基化率为57.1%。结论 abca4和cd200启动子区甲基化改变会影响自身的基因表达量。心衰时DNA启动子区甲基化会发生改变,并且DNA启动子区的甲基化会伴随着基因表达量的改变。
Objective To investigate the effect of genome-wide methylation on myocardial gene expression in patients with heart failure. Methods Totally 8 cases of donor left ventricular myocardium during heart transplantation were selected as normal control group and 14 cases of left ventricular myocardial samples from heart transplant patients undergoing cardiac transplantation were used as experimental group. Genomic DNA was extracted by methylation-based DNA immunoprecipitation combined with methyl Methylated DNA immunoprecipitation-chip (MeDIP-chip) and gene expression microarray for high-throughput rapid screening. Then, two methylated abnormal genes, abca4 and cd200, were obtained after MeDIP-chip pre-screening for sodium bisulfite sequencing PCR. Results The methylation rate of promoter region of abca4 gene was 85.5% in disease group and 91.2% in normal group. The methylation rate of promoter region in disease group was higher than that in normal group The rate of methylation in the promoter region of cd200 gene was 50.5% in HF group and 57.1% in normal group. Conclusion The methylation changes of the promoter region of abca4 and cd200 will affect their own gene expression. DNA damage in heart failure occurs when the methylation of the promoter changes, and DNA methylation in the promoter region is accompanied by changes in gene expression.