云南白族低枸橼酸尿症与VDR基因启动子甲基化水平的关系  

Relevance between hypocitraturia and VDR gene promoter methylation of the Bai nationality in Yunnan province

作　　者：林晓伟 罗钰辉[1] 李静玲 张白羽[1] 柯坤彬[1] 李颢[1] Lin Xiaowei;Luo Yuhui;Li Jingling;Zhang Baiyu;Ke Kunbin;Li Hao(Dept of Urology,the First Affiliated Hospital of Kunming Medical University,Kunming 650032)

机构地区：[1]昆明医科大学附属第一医院泌尿外科,昆明650032

出　　处：《安徽医科大学学报》2023年第4期573-576,共4页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:81460141);云南省高层次卫生技术人才项目(编号:D-2017043);云南省科技厅应用基础研究重点项目(编号:202001AY070001-158)。

摘　　要：目的 研究维生素D受体(VDR)基因启动子甲基化水平与云南白族人群特发性低枸橼酸尿症之间的关联性。方法 选取15例VDR基因单核苷酸多态性位点(SNP shot)rs2228570(FokⅠ)基因型为双显性表达(FF型)的云南白族特发性低枸橼酸尿症患者为实验组,15例尿枸橼酸含量正常的云南白族体检者为对照组。先取两组人员的血液标本,经过亚硫酸盐处理后,通过PCR扩增、T7 DNA聚合酶的体外转录过程得到各RNA产物,经碱基特异性酶切处理后得到对应RNA小片段,最后通过EpiTYPER程序得出每个检测位点的甲基化程度。结果 与对照组[1.261%(0.827%,1.930%)]比较,实验组的VDRⅠ片段甲基化水平[4.136%(1.655%,5.152%)]较高,差异有统计学意义(P<0.001);在VDRⅠ片段上33个CpG位点中,实验组和对照组在CpG-5(F=8.831,P=0.008)和CpG-8(F=16.155,P=0.001)两个位点上的DNA甲基化水平差异有统计学意义。结论 VDR基因启动子甲基化水平升高与云南白族特发性低枸橼酸尿症的发生存在关联性。VDR基因SNP位点FokⅠ为FF型的白族患者VDR基因启动子DNA甲基化水平较正常白族人群显著升高。Objective To investigate the relevance between vitamin D receptor(VDR)gene promoter methylation level and idiopathic hypocitraturia of the Bai nationality in Yunnan province.Methods Fifteen patients with idiopathic hypocitrouria of the Bai nationality in Yunnan province with double dominant expression(FF type)of single nucleotide polymorphism shot(SNP shot)rs2228570(FokⅠ)genotype were selected as the experimental group.Fifteen people of the Bai nationality in Yunnan province with normal content of urinary citric acid were the control group.First,blood samples were taken from both groups.Next,the blood samples were treated with sulfites,RNA products of each sample were obtained by PCR amplification and in vitro transcription of T7 DNA polymerase.Then the corresponding RNA fragments were digested by base-specific enzymes.Finally the degree of methylation at each test site was obtained through the EpiTYPER procedure.Results In the statistical results of DNA methylation level,the methylation level of VDRⅠfragment experimental group was 4.136%(1.655%,5.152%)which was higher than 1.261%(0.827%,1.930%)in the control group,and the difference was statistically significant(P<0.001).Among the 33 CpG sites on VDRⅠfragment,there were significant differences in DNA methylation levels of CPG-5(F=8.831,P=0.008)and CpG-8(F=16.155,P=0.001)between the experimental group and the control group.Conclusion The increased methylation level of VDR gene promoter is related with idiopathic hypocitric of the Bai nationality of Yunnan province.And compared with the normal Bai nationality people,the DNA methylation level of VDR gene promoter significantly increased in the Bai nationality patients with FF type of VDR gene SNP shot FokⅠ.

关 键 词：维生素D受体 基因启动子 DNA甲基化 特发性低枸橼酸尿症 

分 类 号：R691.4[医药卫生—泌尿科学]