全反式维甲酸对川崎病患儿外周血Th17/Treg细胞平衡的影响研究  

The effect of all-trans-retinoic acid on the balance of Th17/Treg in the peripheral blood of patients with Kawasaki disease

作　　者：黎萍[1] 倪维华[2] 潘薇[1] 田婧[1] 郑百红[1] 

机构地区：[1]吉林大学第二医院儿科,吉林长春130041 [2]吉林大学基础医学院免疫学系

出　　处：《中国实验诊断学》2017年第5期781-785,共5页Chinese Journal of Laboratory Diagnosis

基　　金：吉林省科技厅青年基金资助课题(20140520022JH)

摘　　要：目的探讨全反式维甲酸(ATRA)对川崎病患儿外周血Th17/Treg细胞平衡的影响。方法研究纳入10名川崎病患儿为病例组,选择10名健康儿童作为对照组,采取外周静脉血后,无菌分离、制备外周血单个核细胞(PBMC),给予不同浓度ATRA诱导培养PBMC72h后,采用酶联免疫吸附法(ELISA)检测细胞上清液的细胞因子IL-10、IL-17A、TGF-β以及IL-6水平,实时PCR检测RORγt mRNA和FOXP3 mRNA表达水平,流式细胞术检测Th17/Treg占外周CD4+T细胞百分比。结果未加ATRA作用下,病例组Th17细胞比例,转录因子RORγt的mRNA表达水平以及相关细胞因子IL-10、IL-17A均高于对照组(P均<0.01),而Treg细胞比例,FOXP3mRNA表达水平以及相关细胞因子TGF-β、IL-6低于对照组(P均<0.01)。给予不同浓度ATRA作用后,病例组以及对照组Th17比例、IL-10、IL-17A、RORγt的mRNA均有所下降,而Treg细胞比例、FOXP3mRNA表达水平、TGF-β、IL-6均有所上升(P均<0.01)。结论 ATRA在体外可促进川崎病患儿外周血Treg分化,同时抑制Th17分化,改变川崎病患儿外周血Th17/Treg失衡状态,该研究为阐明川崎病的免疫学发病机制及探索新疗法提供实验依据以及思路。Objective To examine the effect of all-trans-retinoic acid（ATRA） on the balance of Th17/Treg in the peripheral blood of patients with Kawasaki disease. Methods Peripheral venous blood samples were obtained from 10 children with Kawasaki disease and 10 healthy children as control. The peripheral blood mononuclear cells （PBMC） were isolated aseptically,and the cells were cultured with different concentrations of ATRA for 72 hours. Plasma levels of Th17-and Treg-related cytokines including IL-17A, IL-10, IL-6, TGF-β in cell supernatant and mRNA expression levels of RORTt and FOXP3 were tested using ELISA and real time PCR, respectively. Flow cytometry was used to detect the percentage of Th17/Treg in CD4＋ T ceils. Results Without effect of ATRA,PBMC from patients with Ka- wasaki disease had higher levels of Th17 percentage,IL-17A, IL-10 and RORγt mRNA （P〈0.01） when compared to the control group. ATRA resulted in a reduction in Th17 percentage,IL-17A,IL-10 and RORγt mRNA （P〈0. 01） and an increase of Treg percentage, IL-6, TGF-β and FOXP3 mRNA in PBMC from patients with Kawasaki disease and control group（P〈0. 01）. Conclusion ATRA could promote Treg differentiation of peripheral blood in children with Kawasaki disease in vitro,and inhibit Th17 differentiation. The Th17/Treg imbalance in peripheral blood of children with Kawasaki disease was changed by ATRA in vitro. Our findings provide further evidence for the immune pathogenesis and therapy of Kawasaki disease.

关 键 词：川崎病 维甲酸 Th17细胞 TREG细胞 细胞因子 

分 类 号：R725.4[医药卫生—儿科]