新诊断1型糖尿病患者外周血自然杀伤细胞的变化特点  

Changes of natural killer cells in peripheral blood in patients with newly-diagnosed type 1 diabetes mellitus

作　　者：杨明兰[1] 章晓芳[2] 王莉[3] 高捷[1] 王博凯[1] 洪洁[1] 李小英[1] 王卫庆[1] 宁光[1] 顾卫琼[1] 

机构地区：[1]上海交通大学医学院附属瑞金医院内分泌代谢病科上海市内分泌代谢病临床医学中心上海市内分泌代谢病研究所,200025 [2]杭州市第一人民医院 [3]复旦大学附属中山医院内科

出　　处：《中华内分泌代谢杂志》2012年第6期455-459,共5页Chinese Journal of Endocrinology and Metabolism

基　　金：上海市科委课题（11ZR1432000）;卫生行业科研专项项目（201202008）

摘　　要：目的探讨新诊断1型糖尿病患者外周血自然杀伤细胞数目和功能的变化情况。方法收集2009年到2011年瑞金医院收治新诊断1型糖尿病患者外周全血标本43份，同时收集性别、年龄匹配的正常对照健康志愿者外周全m标本21份，2型糖尿病对照组14例．Ficoll方法分离外周血单个核细胞（PBMC），流式细胞术（FACS）测定PBMC主要细胞亚群比例，流式细胞分选仪分离正常人和1型糖尿病患者自然杀伤细胞。实时荧光定量PCR检测自然杀伤细胞激活的主要效应分子γ-干扰素（IFN-γ）、perforin及自然杀伤细胞激活性受体NKp46、NKp30基因mRNA表达水平。结果与正常对照相比，自然杀伤细胞是1型糖尿病患者所有细胞亚群中改变最明显的一群细胞，细胞数目明显降低[（102±86）／μl对（355±264）μl，P〈0．01]；与正常对照自然杀伤细胞相比，1型糖尿病患者的自然杀伤细胞NKp46、perforin基因表达明显下凋（P〈O．05），NKp30、IFN-μ基因在1型糖尿病患者的自然杀伤细胞中表达无明显改变（P〉0．05）。结论新诊断1型糖尿患者存在自然杀伤细胞数目和功能明显下降，自然杀伤细胞介导的免疫功能障碍可能在1型糖尿病的病程中发挥着非常重要的作用。Objective To investigate the changes in number and function of natural killer （NK） cells in patients with newly-diagnosed type 1 diabetes mellitus. Methods Cell counting was performed in peripheral blood mononuclear cell （ PBMC ） subsets in 43 cases with newly diagnosed type 1 diabetes （ T1D ）, 14 cases with newly diagnosed type 2 diabetes（ T2D ） and 21 eases of normal controls by flow cytometry sorting. And then, isolating and collecting NK cells were performed in T1D patients and normal controls. Real time PCR was performed to evaluate the mRNA expression levels of NK cell activity related genes IFN-~/, perforin, NKFA6, and NKp30 in NK cells. Results Compared to normal controls, both the proportion and the absolute counting of NK cells in PBMC from patients with TID were significantly decreased [ （ 102±86）/μl vs （355±264）/μl,P〈0.01 ], while only the proportion of CD4＋cell were slightly increased（ P 〈 0.05 ）. No statistical difference was observed regarding CD8 ＋ T cells （ P 〉 0.05 ）. mRNA expression levels of NK cell activity related genes perforin and NKp46 in NK cells were remarkably down-regulated （ P〈0.05 ） , while IFN-γ and NKp30 were not changed compared with normal controls. Conclusions The reduced number and functional deficiency of NK ceils may lead to the immune dysfunction in T1D and play an important role in the development of TID.

关 键 词：糖尿病 1型 外周血单个核细胞 自然杀伤细胞 

分 类 号：R587.1[医药卫生—内分泌]