中国汉族2型糖尿病人群中UCP基因单核苷酸多态性与视网膜病变的关联分析

摘要

Background Researches showed that elevatory blood glucose level results in long-term damage of cells and tissue,or metabolic memory phenomenon,and manipulation of hyperglycemic memory is a good approach in the prevention of diabetic complications.However,its mechanism is not clear.It is speculated that the pathogenesis of diabetic retinopathy (DR) in diabetic patients may be associated to related mechanisms.Uncoupling proteins (UCPs) can decrease the production of reactive oxygen species (ROS),which may be related to DR.Objective This study was to explore the association between DR and the single nucleotide polymorphisms (SNPs) of UCP genes in Chinese Han population with type 2 diabetes.Methods A cross-sectional study was performed.This study was approved by Ethic Committee of Affiliated First Hospital of Shanghai Jiao Tong University and complied with Declaration of Helsinki,and written informed consent was obtained from each subject prior to any medical examination.One thousand eight hundreds and seventy-five patients with type 2 diabetes mellitus were enrolled in Xinjing district of Shanghai city by cluster sampling from November 2014 to January 2015.The demographic and medical baseline characteristics,ocular examination and laboratory tests were obtained and periphery blood of 2 ml was collected for extraction of DNA.Eight tag SNPs of UCP1,three tag SNPs of UCP2,and seven tag SNPs of UCP3 were selected as marker locus for the detection of genotype by Sequenom Mass ARRAY.Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry platform were used for genotyping.Hardy-Weinberg equilibrium (HWE) analysis,allele and genotype frequencies,haplotype analysis,and association tests for DR and SNPs were performed by SAS and SHEsis software.Results A total of 530 DR patients were checked out from 1 875 subjects with type 2 diabetes mellitus,with the detection rate of 28.27％.rs660339 locn of UCP2 gene and rs1626521,rs668514 locus of UCP3 gene appeared to have low detectable rates,and the secondary allele base frequency of rs632862 in UCP2 gene was ＜0.01 and rs15763 of UCP3 gene was unmatched with HWE,therefore,these locus analysis was not included.In 13 SNPs locus included in the analysis,only 2 SNPs of UCP1 gene were related to DR.Compared with the non-diabetic retinopathy (NDR) patients,the G allele frequency of rs10011540 was increased (P =0.03,OR =1.31,95 ％ confidence interval[CI] =1.03-1.67,and T allele frequency of rs3811787 was decreased (P=0.04,OR=0.86,95％ CI=0.75-0.99) in DR patients.Genotyping detection showed that the C/C and A/A frequencies of rs3811790 in UCP1 gene were significantly more and C/A frequency was less in DR patients than those in NDR patients (all at P＜0.01).The logistic regression analysis indicated an association of SNPs of rs10011540 and rs3811787 with DR independent from glucose and disease duration.Conclusions The SNPs of rs10011540 and rs3811787 locus in UCP1 gene are associated with DR in Chinese type 2 diabetes patients.%背景 研究发现血糖水平的短期升高可对细胞和组织造成长期损害,这种损伤可能存在代谢记忆现象,合理管理血糖代谢记忆对糖尿病并发症的预防有重要作用,但其机制尚未完全阐明,推测糖尿病患者中糖尿病视网膜病变(DR)的发生可能与相关机制有关.解偶联蛋白(UCPs)可减少线粒体活性氧(ROS)的生成,可能与DR发病相关. 目的 探讨中国汉族2型糖尿病人群中DR与UCP基因单核苷酸多态性(SNPs)之间的关系.方法 采用横断面研究方法和整群抽样法,于2014年11月至2015年1月在上海市新泾社区对1 875例确诊为2型糖尿病的患者进行流行病调查,收集受检者的基本信息、眼科检查和血生物化学检验结果,采集每例患者的全血2 ml以提取DNA.采用Sequenom平台将UCP1基因的8个SNPs位点、UCP2基因的3个SNPs位点及UCP3的7个SNPs位点选为标记位点以检测基因型,采用SAS和SHEsis软件计算Hardy-Weinberg平衡、碱基型和基因型频率,评估各位点SNPs与DR之间的关系. 结果 受检的1 875例2型糖尿病患者中530例患DR,占28.27％.UCP2基因的rs660339位点和UCP3基因的rs1626521位点、rs668514位点的检出率低,UCP2基因rs632862位点次要等位碱基频率＜0.01,UCP3基因的rs15763位点不符合Hardy-Weinberg平衡,故均不纳入分析.在纳入分析的13个SNPs位点中,仅有UCP1基因的2个SNPs位点与DR发病有关,其中与非糖尿病视网膜病变(NDR)患者比较,DR患者rs10011540的G碱基频率增加[P=O.03,OR=1.31,95％可信区间(C1)=1.03～1.67],rs3811787的T碱基频率下降(P=0.04,OR=0.86,95％ CI=0.75 ～0.99).基因型分析发现,DR患者UCP1基因的rs3811790位点纯合子C/C和A/A频率明显多于NDR患者,杂合子C/A频率少于NDR患者,差异均有统计学意义(P＜0.01).Logistic回归分析提示,在排除了血糖水平和糖尿病病程的影响因素后,rs10011540和rs3811787位点SNPs仍是DR发病的独立影响因素.结论 中国汉族2型糖尿病患者UCP1基因rs10011540和rs3811787位点SNPs与DR发病相关.