基于人2型糖尿病易感SNP位点筛查食蟹猴2型糖尿病易感SNP标记

Screening of genetic markers based on human type 2 diabetes mellitus(T2DM) susceptibility loci in a Cynomolgus T2DM model

目的 2型糖尿病(type 2 diabetes mellitus,T2DM)及其并发症的迅速蔓延已经成为严重威胁人类健康的世界性难题。食蟹猴T2DM模型可以模拟人类T2DM发病进程,这对T2DM的预防,治疗和新药开发具有重要意义。本研究以37个与人密切关联的T2DM易感位点为基础,探讨对应食蟹猴同源区域的这些位点与83个食蟹猴个体之间的关联性。方法先采用DNA池法初筛,然后对食蟹猴个体进行直接聚合酶链式反应(polymerase chain reaction,PCR)扩增测序,并利用DNAStar软件分析碱基之间的差异性,最后采用方差分析和F检验计算等位基因间的频率,并采用SAS的GLM模型进行基因型与空腹血糖值和糖化血红蛋白值之间的关联分析。结果在自发性T2DM组和对照组中,单核苷酸多态性(single nucleotide polymorphisms,SNP)位点SNPmf-66A、SNPmf-66B、SNPmf-34A、SNPmf-34B、SNPmf-34C、SNPmf-56A、SNPmf-56B和SNPmf-56C的等位基因频率存在差异显著性(P<0.05),同时SNPmf-66A、SNPmf-66B、SNPmf-34A、SNPmf-34B和SNPmf-34C与空腹血糖之间也存在显著性关联性(P<0.05)。结论 SNPmf-66A、SNPmf-66B、SNPmf-34A、SNPmf-34B、SNPmf-34C可能对食蟹猴T2DM动物模型构建具有重要意义。

Objective Type 2 diabetes mellitus （ T2DM） as a common disease around the world becomes a great threat to the health of human beings.The cynomolgus T2DM model, which preferably simulates human T2DM onset and progress, can be beneficial to the drug development and clinical treatment.In the present study, 37 of T2DM-susceptibility SNPs and the extended genome sequences were used to obtain corresponding SNPs in the T2DM cynomolgus monkeys. Methods Firstly, DNA pool screening was conducted.Then, using polymerase chain reaction to amplify and to sequence the cynomolgus homologous sequences.Using DNAStar software to analyze the differences between bases.Finally, we used analysis of variance and F test to calculate the frequency of alleles.We also used the GLM models of SAS software to analyze the association of genotype with fasting plasma glucose and glycosylated hemoglobin.Results SNP661A,SNP661B, SNP343A, SNP343B, SNP343C, SNP565A, SNP565B and SNP565C were found to have a significant difference of allele frequencies between spontaneous cases and controls.Conclusions The findings of this study suggest that SNP661A, SNP661B, SNP343A, SNP343B and SNP343C may play an important role in the establishment of cynomolgus T2DM models.