正常人与系统性红斑狼疮患者白细胞GPI-PLD基因外显子14的碱基变异分析  

Survey of GPI-PLD gene exon14 variations in healthy person and patients with systemic lupus erythematosus

作　　者：黄河[1] 杨智英[1] 唐建华[1] 禹虹[1] 王依丹[1] 向新颖[1] 

机构地区：[1]中南大学生物化学教研室

出　　处：《中国现代医学杂志》2005年第19期2938-2942,共5页China Journal of Modern Medicine

基　　金：国家自然科学基金资助项目(39970315;30271456)

摘　　要：目的检测湖南地区汉族系统性红斑狼疮(SLE)患者与正常人外周血白细胞糖基化磷脂酰肌醇特异性磷脂酶D(GPI-PLD)基因外显子14区的碱基变异、白细胞GPI-PLD活性及二者关系。方法PCR-SSCP结合DNA测序技术进行碱基变异分析,利用自制具完整GPI结构的胎盘碱性磷酸酶做底物通过TX-114分相,定量检测白细胞中GPI-PLD活性。结果湖南地区汉族48名SLE患者与96名正常人外周血白细胞GPI-PLD基因外显子14区均存在4处碱基变异,外显子第1257位C→T密码子ATC(Ile)→ATT(Ile)、1298位T→C密码子CTG(Leu)→CCG(Pro)、1218位C→A密码子AGC(Ser)→AGA(Arg)、1257位C→A密码子ATC(Ile)→ATA(Ile);而在1142位C→G密码子AAC(Asn)→AAG(Lys)、1222位C→T密码子CCC(Pro)→TCC(Ser)、1207位C→T密码子CCA(Pro)→TCA(Ser)、1278位G→T密码子CTG(Leu)→TCC(Leu)仅存在于SLE人群;经SSCP检出阳性率分别为正常人20.83%,SLE患者22.91%。检测48例SLE患者外周血白细胞GPI-PLD活性(转化底物百分率),发现SLE患者酶活性较正常人(96名)显著性增高,增高的百分率为38.01%。但与碱基变异位点无显著相关性。结论湖南地区汉族系统性红斑狼疮患者与正常人外周血白细胞GPI-PLD基因外显子14均存在碱基变异,SLE患者白细胞GPI-PLD酶活性较正常人显著增高。[Objective] To analyze the various frequency of glycosylphosphatidyhnositol - specific phospholipase D （GPI-PLD） gene Exon I4, GPI-PLD activity of leucocyte in peripheral blood and their relationship in systemic lupus erythematosus（SLE） patients of Han nationality in Hunan Chinese. [Methods] The variations of GPI-PLD gene were analyzed by PCR-SSCP and sequencing. GPI-PLD activities were determined by using GPI-anchored placental alkaline phosphatase （PLAP）as substrate and tritonX-114 partioning. [Results] There were four variations in the coverage of GPI-PLD gene Exon I4 of both 48 SLE patients＇ and 96 healthy persons＇. The codons of variation were： 1 257C→T ATC（Ile）→ ATT（Ile）, 1 298 T→ CTG（Leu）→CG（Pro）, 1 218 C→A AGC（Ser）→ AGA （Arg）, I 257 C→A ATC（Ile）→ATA （lie）. In addition, the variation of I 142 C→ AAC（Asn）→AAG （Lys）, 1 222 C→T CCC（Pro）→TCC （Ser）, I 207 C→T CCA（Pro）→TCA （Ser）, 1 278 G→T CTG（Leu）→TCC （Leu） was only in SLE patients＇. The total various frequency in SLE patients and healthy person, which was determined by SSCP, was 22.92 percent and 20.83 percent. On the basis of the percentage of GPI-anchored PLAP conversion, the leucocyte GPI- PLD activities of total 48 SLE patients were measured. As compared with 96 healthy persons as control group, the leukocyte GPI - PLD activities of SLE patients were significantly raised, but GPI-PLD activities were not correlated with mutation sites significantly. [Conclusions] There are base mutations of leukocyte GPI-PLD gene in peripheral blood which belongs to healthy persons and SLE patients of Han nationality in Hunan Chinese. The leukocyte GPI- PLD activeties of SLE patients also significantly increase than those of healthy group.

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 系统性红斑狼疮 

分 类 号：R593.24[医药卫生—内科学]