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目的:慢性阻塞性肺疾病患者白介素-6基因的提取及668C/G位点基因分型。慢性阻塞性肺疾病(chronic obstructive pulmonary disease,COPD)是一个复杂的全身性疾病,其发病是多基因遗传、环境等因素共同作用的结果。它的特点是既有肺的局部炎症也有全身性炎症。许多研究提示COPD患者出现血循环中的炎症细胞活化和炎症细胞因子水平增高。方法:从COPD患者外周血提取基因组DNA,微量核酸测定仪检测核酸浓度,通过聚合酶链式反应(polymerasechainreaction,PCR)扩增目的基因白介素-6基因片断,用胶回收的方法回收和纯化白介素-6基因,并用限制性内切酶剪切白介素-6基因,检测IL-6基因668C/G位点的基因型。结果:COPD患者668C/G位点基因分型可分为CC,CG,GG基因型。结论:成功进行了IL-6基因668C/G位点的基因型分析,为之后进一步进行多态性分析研究奠定基础。
OBJECTIVE: To extract IL-6 gene and genotypes of 668C / G locus in patients with chronic obstructive pulmonary disease. Chronic obstructive pulmonary disease (COPD) is a complex systemic disease, the onset of which is the result of a combination of multiple genetic and environmental factors. It is characterized by both local and systemic lung inflammation. Many studies have suggested that patients with COPD appear to have increased inflammatory cell activation and increased levels of inflammatory cytokines in the blood circulation. Methods: Genomic DNA was extracted from the peripheral blood of patients with COPD. The concentration of nucleic acid was detected by micro-acid detector. The target gene interleukin-6 gene fragment was amplified by polymerase chain reaction (PCR), and the IL-6 gene was recovered and purified by gel filtration. 6 gene and cut IL-6 gene with restriction endonuclease to detect the genotype of 668C / G of IL-6 gene. Results: Genotypes of 668C / G in COPD patients were classified into CC, CG and GG genotypes. Conclusion: The genotype analysis of IL-6 gene 668C / G locus was successfully carried out, which laid the foundation for the further study of polymorphism.