2型糖尿病患者外周血CD14^+CD16^+单核细胞水平及巨噬细胞STAT3蛋白表达的研究  

Mean fluorescence intensity of CD14^+CD16^+ monocyte from patients with type 2 diabetes mellitus and the protein expression of STAT3 in macrophages

作　　者：杨孟雪[1] 杨波[1] 李思成[1] 甘华[2] 李显文[1] 晏永慧[1] 

机构地区：[1]遵义医学院附属医院内分泌科,563003 [2]重庆医科大学附属第一医院肾内科

出　　处：《中国糖尿病杂志》2015年第1期59-63,共5页Chinese Journal of Diabetes

基　　金：贵州省科技攻关项目[黔科合SY字(2012)3116号];贵州省科学技术基金项目[黔科合J字LKZ(2013)53号];贵州省科学技术基金项目[黔科合J字LKZ(2012)28号];遵义医学院博士启动基金(F-588)

摘　　要：目的研究T2DM患者外周血CD14+CD16+单核细胞数和血清IL-6、TGF-β的水平,及患者血清和IL-6孵育的THP-1单核细胞源性巨噬细胞STAT3、p-STAT3的蛋白表达,以了解炎症性免疫反应在T2DM大血管病变中的可能作用。方法对42例T2DM(T2DM组)患者和35名健康体检者(NC组)采用外周血流式细胞术检测外周血单核细胞CD14+CD16+的表达,并分离其外周血血清。采用ELISA检测血清中细胞因子IL-6及TGF-β的浓度,免疫比浊法检测血清中高敏C反应蛋白(hsC-RP)水平,血清和IL-6孵育THP-1单核细胞源性巨噬细胞,采用Western blot检测其STAT3和p-STAT3的蛋白表达。结果 T2DM组外周血CD14+CD16+单核细胞数高于NC组(P<0.01),且与血清hsC-RP和IL-6水平呈正相关(r=0.462、0.495,P<0.01),与TGF-β水平无相关性(P>0.05)。T2DM组24hUAlb水平、大血管病变发病率及THP-1单核细胞源性巨噬细胞p-STAT3的蛋白表达均高于NC组(P<0.01)。结论 T2DM患者体内可能存在单核/巨噬细胞功能异常,其可能参与了T2DM患者体内免疫炎症反应,从而导致了T2DM及其血管病变的发生发展,作用机制可能与STAT3信号通路的激活有关。Objective To characterize the expression of CD14^＋CD16^＋monocytes and STAT3 and phosphorylated（p）-STAT3 protein expressions monocyte-derived macrophages in type 2diabetes（T2DM）patients.And to further explore the potential effects of inflammatory immune response in T2 DM macrovascula. Methods Forty-two patients with T2 DM,and thirty-five healthy subjects were enrolled for the determination of CD14＋CD16＋monocyte by using peripheral blood flow cytometry.Serum IL-6and TGF-βlevels were assessed by ELISA.Serum high-sensitivity C-reactive protein（hsC-RP）level was determined by immunoturbidimetry.THP-1monocyte-derived macrophages were incubated for 12 hwith serum from healthy control and T2 DM patients,and were stimulated by IL-6for 4h.THP-1 monocytederived macrophages were collected to detect STAT3 and phosphorylated（p）-STAT3 protein by using western blot. Results Compared to normal control,T2 DM patients had a significantly higher levels of CD14^＋CD16^＋fluorescence intensity,serum hsC-RP and IL-6level.There were positive correlation of CD14＋CD16＋monocytes quantity with serum hsC-RP and IL-6（r=0.462、0.495,P〈0.01）,without correlation between serum TGF-βand CD14^＋CD16^＋monocytes quantity.Compared to the control,T2 DM patients had a significantly higher 24 hUAlb and macrovascular disease incidence（all P〈0.05）.After incubated with sera deriving from the groups and stimulated by IL-6,the THP-1 monocyte-derived macrophages showed a significant upregulation in p-STAT3 protein expression（P〈0.01）. ConclusionThese findings suggest that the function disturbance in monocytes occurs in T2 DM.This dysfunction may be related to the inflammatory immune response in patients with T2 DM,which results in the occurrence and development of T2 DM and macrovascular disease.The mechanism may be related to activation of STAT3 signaling pathway.

关 键 词：糖尿病 2型 CD14^+CD16^+单核细胞 STAT3 白介素-6 

分 类 号：R587.1[医药卫生—内分泌]