摘要目的 观察中国汉族人群中血管内皮原生型一氧化氮合酶(ecNOS)基因的多态性与糖尿病视网膜病变(DR)之间的相关性.方法 166例临床确诊为非胰岛素依赖型糖尿病患者为病例组,选取无糖尿病、高血压、肾病,年龄40岁以上,个体间无血缘关系的85例白内障、骨折患者和健康体检者为正常对照组.病例组患者根据有无视网膜病变和荧光素眼底血管造影检查结果分为无DR组、非增生期DR组(BDR组)和增生期DR组(PDR组).病例组和正常对照组受检者均为汉族.抽取外周静脉血,提取DNA.采用聚合酶链反应(PCR)检测ecNOS基因第四内含子中27个碱基对(bp)重复的多态性.结果 PCR产物测序结果经检索GeneBank,扩增序列与GeneBank中ecNOS基因相应区域的序列相一致,显示在汉族人群中同样存在27个bp的重复序列,等位基因b重复5次,等位基因a重复4次.PCR检测结果显示,在正常对照组和无DR组、BDR组、PDR组中均存在2种等位基因和3种基因型.正常对照组中基因型bb、ab、aa分别为80.0%、16.5%、3.5%、无DR组分别为77.2%、13.9%、8.9%、BDR组分别为80.5%、17.1%,2.4%,PDR组分别为78.3%、13%、8.7%;正常对照组、无DR组、BDR组、PDR组两两比较,等位基因频率(χ~2=1.841)和基因型频率(χ~2=3.847)均无统计学意义(P>0.5).Logistic回归分析结果也显示DR与ecNOS基因重复多态性无关.结论 中国汉族人群ecNOS基因第四内含子存在27个bp重复的多态性,但可能与非胰岛素依赖型糖尿病患者视网膜病变无关.

abstractsObjective To observe the relationship between endothelial constitutive nitric oxide synthase (ecNOS) genetic polymorphism and diabetic retinopathy(DR)of non insulin-dependent diabetes mellitus (NIDDM) patients of the Han nationality. Methods A total of 166 patients who clinical diagnosed with NIDDM as case group, 85 cases of patients (cataract or fracture) and healthy subjects without diabetes, hypertension and kidney disease, over 40 years old of age and without consanguinity between each other were selected as normal control group. Case group were divided into non-DR (NDR) group, non-proliferative-DR (BDR) group and proliferative-DR (PDR) group according to the result of fundus fluorescein angiography. Case group and normal control group subjects all were Han nationality. DNA was extracted from peripheral venous blood; the fourth 27 base pairs (bp) repeat polymorphism of ecNOS gene by was measured by polymerase chain reaction (PCR). Results The 27 bp repeat sequences within the ecNOS gene present in the Han nationality, allele b repeat 5 times, alleles a repeat 4 times. PCR results showed that there are 2 alleles and 3 genotypes in normal control, NDR, BDR and PDR group. The frequency of genotype bb、ab、aa were 80%, 16.5%, 3.5% in normal subjects; 77.2%, 13.9%, 8. 9% in NDR group; 80. 5%, 17. 1% ,2.4% in BDR group; 78. 3%, 13%, 8.7% in PDR group,respectively. The allele frequency (χ~2=1.841 ) and gene frequency (χ~2 = 3. 847) were not statistically significant (P>0. 5) in normal control, NDR, BDR and PDR group. Logistic regression analysis showed that there is no relation between DR and ecNOS duplicated gene polymorphism. Conclusions There is 27 bp repeated polymorphism in 4th intron of ecNOS gene, which may not be associated with the DR of NIDDM in the Han nationality.