正常人与系统性红斑狼疮患者白细胞GPI-PLD基因多态性分析  

Survey of GPI-PLD Gene Polymorphisms in Healthy Persons and Patients With Systemic Lupus Erythematosus

作　　者：禹虹[1] 唐建华[1] 王依丹[1] 张晓杰[1] 顾善兰[1] 王成红[2] 

机构地区：[1]中南大学湘雅医学院生物化学教研室,湖南长沙410078 [2]中南大学湘雅三医院

出　　处：《中国医师杂志》2003年第6期724-728,共5页Journal of Chinese Physician

基　　金：国家自然科学基金资助课题 (No .39970 31 5)

摘　　要：目的　检测湖南地区汉族系统性红斑狼疮 (systemiclupuserythematosus ,SLE)患者外周血白细胞糖基化磷脂酰肌醇特异性磷脂酶D (glycosylphosphatidylinositol-specificphospholipaseD ,GPI -PLD)基因外显子 1及外显子 2 5多态性、白细胞GPI -PLD活性及二者关系。方法　聚合酶链式反应与单链构象多态性分析 (PCR -SSCP)、测序技术进行多态性分析 ,利用自制具完整糖基化磷脂酰肌醇 (GPI)结构的胎盘碱性磷酸酶 (PLAP)做底物通过TX -114分相 ,定量检测白细胞中GPI -PLD活性。结果　湖南地区汉族 62名SLE患者与 10 9名正常人外周血白细胞GPI -PLD基因外显子 1区均存在 14处核苷酸改变 :转录起始点上游 -2 8A→C、-2 1C→T、-14G→A、-13G→C、-9T→C、-8G→C、-7G→C、+4T→G、密码子 14TTG→AAG (Leu14Lys)、密码子 17CTC→GTC(Leu17Val)、密码子 2 0AGA→AAA (Arg2 0Lys)、密码子 2 1GGT→GGG (Gly2 1Gly)、密码子 3 0GTA→ATA (Val3 0Ile) ;另转录起始点上游 -2 4C→T仅存在于SLE人群中 ,此变异在两组人群中具有显著性差异 ;该基因外显子 2 5 ,即 +60 496G→A均存在于两组人群中。经SSCP检出阳性率分别为正常人 3 3 0 3 %,SLE患者 2 5 81%。检测 62例SLE患者外周血白细胞GPI -PLD活性 (转化底物百分率 )Objective To analyze the polymorphisms of glycosylphosphatidylinositol-specific phospholipase D (GPI-PLD) gene Exon1 and Exon25,GPI-PLD activity of leucocyte in peripheral blood and their relationship in systemic lupus erythematosus patients of Han nationality in Hunan Chinese.Methods The polymorphisms were analyzed by PCR-SSCP and sequencing. GPI-PLD levels were determined by using glycosylphosphatidylinositol (GPI) anchored placental alkaline phosphatase (PLAP) as substrate and triton-X114 partioning.Results There were fourteen variations in the coverage of GPI-PLD gene Exon 1 of both 109 healthy persons' and 62 systemic lupus erythematosus patients',including seven variations at transcriptional start site upstream and seven variations at Exon1. The codons of variation were 14TTG→AAG (Leu14Lys),17CTC→GTC (Leu17Val),20AGA→AAA (Arg20Lys),21GGT→GGG (Gly21Gly),30GTA→ATA (Val30Ile),which leaded to four missense mutations and one synonymous mutation. In addition, the frequency of the variation at transcriptional start site upstream -24 C→T was significantly different between healthy control and systemic lupus erythematosus patients. At meanwhile, there was a variation in Exon25, the +60496 G→A, between two groups. The total various frequency, which was determined by SSCP, of healthy person was 33 03 percent and systemic lupus erythematosus patient was 25 81 percent. On the basis of the percentage of GPI-anchored PLAP conversion, the leucocyte GPI-PLD activities of total 62 systemic lupus erythematosus patients were measured. As compared with 109 healthy persons as control group, the leukocyte GPI-PLD activities of systemic lupus erythematosus patients were significantly raised, but GPI-PLD activities were not correlated with polymorphism sites significantly.Conclusions The leukocyte GPI-PLD gene in peripheral blood, which belongs to healthy persons and systemic lupus erythematosus patients of Han mationality in Hunan Chinses,is polymorphism.The frequency of transcriptional start site upstream -24

关 键 词：糖基化磷脂酰肌醇特异性磷脂酶D 单核苷酸多态性 多态性 系统性红斑狼疮 

分 类 号：R593.241[医药卫生—内科学]