哮喘病患者TNF-α基因多态性及表达与内皮损伤的研究  

Association of the TNF-α gene of G-308A genotypes and mRNA expression with vascular endothelial cells functional disorder in asthma patients

作　　者：贾少丹[1] 张为忠[1] 纪霞[1] 李靖[1] 赵明[1] 王海燕[1] 

机构地区：[1]青岛市市立医院青岛市常见病重点实验室哮喘病实验室,山东青岛266071

出　　处：《中国现代医学杂志》2013年第21期39-44,共6页China Journal of Modern Medicine

基　　金：青岛市市南区科技发展计划项目(No:2009-4-31-GG);山东省医药卫生科技发展计划项目(No:2009-17);青岛市外国专家局引进国外技术;管理人才项目(No:QD20100002)

摘　　要：目的探讨哮喘病患者TNF-α-308位点基因型分布及mRNA表达对其血管内皮损伤的影响。方法选择3个月内未全身使用过激素及茶碱的哮喘病患者112例,根据病情严重程度分为轻度哮喘组和中重度哮喘组,健康志愿者60例为对照组。采用聚合酶链反应-限制性片段长度多态性分析方法测定TNF-α-308位点多态性,实时荧光定量PCR法检测其外周血单个核细胞中TNF-αmRNA表达水平。同时测定血清中总IgE、TNF-α、CRP、PAI-1、ET-1及一氧化氮水平,测定颈动脉内中膜厚度（CIMT）和肺功能。结果①172名研究对象G-308A TNF-α基因型分布符合Hardy-Weinberg平衡;②中重度哮喘组A等位基因频率与对照组比较有显著性差异（P〈0.05）;③哮喘组GA＋AA基因型组与GG基因型组比较,HOMA-IR、IgE、CRP、TNF-α、PAI-1和ET-1水平明显升高（P〈0.01）,TNF-αmRNA表达、CIMT明显增加（P〈0.01）,一氧化氮水平、FEV1占预计值（%）和FEV1/FVC（%）明显下降（P〈0.01）;④Pearson相关分析结果显示,TNF-αmRNA与TNF-α、CRP、HOMA-IR、PAI-1、ET-1、CIMT呈正相关（r分别为0.631、0.367、0.338、0.243、0.327、0.347,P〈0.01）,与FEV1占预计值（%）、FEV1/FVC、NO呈负相关（r为-0.326、-0.393、-0.199,P〈0.05~0.01）;⑤多因素回归分析显示CIMT与ET-1、HOMA-IR、TNF-αmRNA表达、PAI-1呈正相关（t=1.966~3.788,P〈0.05~0.01）,与FEV1占预计值（%）呈负相关（t=3.181,P〈0.01）。结论哮喘病患者TNF-α308 A携带者的基因型与哮喘及血管内皮损伤相关。提示TNF-α308 A携带可能与哮喘伴发心脑血管危险因素有关。[ Objective ] To study the relationship between TNF-α Gene G-308A genotype, mRNA expression and endothelial cell function in asthma patients. [Methods] We recruited 112 asthmatic patients without systemic use of steroid and theophyline for 3 months, who were separated into mild and moderate/severe asthmatic groups according to the severity of the disease. Genomie DNA was extracted from peripheral blood mononuclear cells of the asthmatics and 60 healthy controls. Single nucleotide polymorphism in the TNF-α gene promoter -308 was genotyped using PCR-RFLP. TNF-α gene expression was analyzed by qRT-PCR. Serum levels of total IgE, TNF-α, CRP, PAI-1, ET-1, NO, Carotid artery intima-media thickness （CIMT） and lung function were examined. [Results] The distributions of TNF-α G-308A genotypes from all individual samples satisfied the Hardy-Weinherg Equilibrium（P〉0.05）. There is significant difference for allele A frequency between severe asthmatics and controls （P 〈0.05）. In asthma groups, HOMA-IR, IgE, CRP, TNF-α, PAI-1, ET-1, TNF-α mRNA expression and CIMT were higher （P 〈 0.01）, but FEV1%, FEV1/FVC% of lung function and serum NO were lower （P 〈0.01） in genotypes GA＋AA group compared with genotype GG group. We found that there are positive correlations between TNF-α mRNA expression and serum levels of TNF-α, CRP, HOMA-IR, PAI-1, ET-1, CIMT （r =0.631, 0.367, 0.338, 0.243, 0.327, 0.347 respectively, P 〈0.01）, however there are negative correlations between TNF-α mRNA expression and FEV1%, FEV1/FVC%, serum NO （r =-0.326, -0.393, -0.199 respectively, P 〈0.05-0.01）. Multiple variable linear stepwise regression showed that CIMT was positively related to ET-1, HOMA-IR, TNF-α mRNA expression, PAI-1 （t = 1.966±3.788, P 〈0.05±0.01）, negatively related to FEV1%（t =3.181, P 〈0.01）. [ Conclusions ] Our data suggested that the TNF-α genotype 308 A was related to the damage of vascular endothelial function in asthma patients, which indicated genotype 308A is a potenti

关 键 词：哮喘 TNF-Α基因多态性 MRNA表达 血管内皮损伤 

分 类 号：R562.25[医药卫生—呼吸系统]