Klotho基因3个位点单核苷酸多态性与冠心病相关性研究

Correlation between single nucleotide polymorphisms at three sites of Klotho gene with coronary heart disease

目的:通过研究克老素(Klotho,KL)基因3个单核苷酸多态性,揭示其与冠心病(Coronary heart disease,CHD)的相关性。方法:随机选取CHD患者317例为实验组,同期随机选取正常健康人301例正常对照组。提取外周血白细胞基因组DNA,应用单一等位基因特异性引物PCR技术检测KL基因3个SNP位点的基因型,分析其各等位基因与CHD的相关性。结果:CHD组KL G-395A SNP AA等位基因频率较正常对照组增高,差异有统计学意义(P<0.05);按性别分层研究发现CHD男性亚组AA等位基因频率较正常对照组增高(P<0.05)。KL C370S SNP CC等位基因频率较对照组增高(P<0.05),而其CS等位基因频率较对照组低(P<0.05);按性别分层研究发现,CHD男性亚组CC频率较对照组偏高,差异有统计学意义(P<0.05),而其CS频率较对照组降低,差异有统计学意义(P<0.05)。未发现CHD组F-352V基因型多态性FF、FV和VV频率与对照组比较差异有统计学意义(P>0.05)。组合分布研究发现:CHD组KL G-395A+F352V+C370C、KL A-395A+F352V+C370C与KL A-395A+F352V+C370S组合基因型分布频率较正常对照组明显增高(P<0.05),而其KL G-395A+F352V+C370S组合基因型分布频率较正常对照组低(P<0.05)。按性别分层研究发现在CHD组男性患者中KL G-395A+F352V+C370C、KL A-395A+F352V+C370C组合基因型频率亦较正常对照男性亚组明显增高(P<0.05),而KL G-395A+F352V+C370S组合基因型分布频率则较正常对照男性亚组低(P<0.05);同时还发现在CHD女性亚组患者KL A-395A+F352V+C370S组合分布基因型分布频率较正常对照女性亚组明显增高(P<0.05)。结论:研究发现KL基因型G-395A SNP AA男性携带者易患CHD,而KL C370S SNP突变CS杂合子男性携带者则不易患CHD。组合分布研究还发现KL G-395A+F352V+C370C、KL A-395A+F352V+C370C男性携带者与KL A-395A+F352V+C370S女性携带者易患CHD,而KL G-395A+F352V+C370S男性携带者则不易患CHD。

Objective：To investigate single nucleotide polymorphisms（SNPS） at three sites of the Klotho（KL） gene and to analyze their correlation with coronary heart disease（CHD）.Methods：Totally 317 patients were randomly selected as CHD group and 301 healthy people as normal controls.Genomic DNA of leukocytes was extracted from venous blood and the single allele specific primer PCR technique（SASP-PCR） was used to detect allelic genotypes at three SNPS sites of KL gene in order to analyze their relationship with CHD.Results：The frequency of allele AA genotype of KL G-395A in CHD group was obviously higher than that in normal control group（P〈0.05）.According to the results of sex stratification analysis,the frequency of allele AA genotype in the CHD male subgroup was higher than that in normal control group（P〈0.05）.The frequency of allele CC genotype of KL C370S was higher than that in normal control group（P〈0.05）,but the frequency of allele CS genotype was lower than that in normal control group（P〈0.05）.According to the results of sex stratification analysis,the frequency of allele CC genotype in CHD male subgroup was higher than that in normal control group（P〈0.05）,but the frequency of allele CS genotype was lower than that in normal control male subgroup（P〈0.05）.There was no significant difference in polymorphic FF,FV and VV frequency in the F-352V gene between CHD group and normal control group（P 〉0.05）.According to the results of combination genotype analysis,the allele genotype frequency of KL G-395A＋F352V＋C370C,KL A-395A＋F352V＋C370C and KL A-395A＋F352V＋C370S in CHD group was higher than that in normal control group（P〈0.05）,but the allele genotype frequency of KL G-395A＋F352V＋C370S was lower than that in normal control group（P〈0.05）.According to the results of sex stratification analysis,the allele genotype frequency of KL G-395A＋F352V＋C370C,KL A395-A＋F352V＋C370C was higher than that in normal control male subgroup（P〈0.05）,but the allele genotype frequency of KL G-395A＋F352V＋C370S was lower than that in normal control male subgroup（P〈0.05）.The allele genotype frequency of KL A-395A＋F352V＋C370S in CHD female subgroup was obviously higher than that in normal control female subgroup（P〈0.05）.Conclusions：This study shows that the male patients who carried with allele genotype KLG-395A SNP AA have higher risk in suffering from CHD.The male patients with allele CS genotype of KL C370S have lower risk in suffering from CHD.This study also finds that the male patients who carried with KL G395-A＋F352V＋C370C,KL A-395A＋F352V＋C370C and the female patients who carried with KL A-395A＋F352V＋C370S have higher risk in suffering from CHD,however,the male patients with KL G-395A＋F352V＋C370S have lower risk in suffering from CHD.