摘要
目的探讨CD4+CD25+Foxp3+Treg/CD4+IL-17A+Th17细胞及相关细胞因子在Graves眼病中的作用及机制。方法根据临床活动性评分(CAS)将未经治疗的Graves眼病患者分为活动性GO组(AGO组)15例(CAS≥3),非活动性GO组(NGO组)15例(CAS<3);同时选取未经治疗的同期非突眼Graves病患者(GD组)15例及健康对照组(C组)15例。收集各组病人外周静脉血,采用流式细胞计数检测Treg、Th17细胞比例的变化;采用荧光定量PCR(RT-PCR)检测Treg特异性转录因子Foxp3及Th17特异性转录因子RORγt的表达情况;使用双抗体夹心酶免疫吸附法(ELISA)检测外周血血清中Treg相关细胞因子TGF-β、IL-10、IL-35以及Th17相关细胞因子IL-17A、IL-23、IL-6的表达变化。结果 GD组、NGO组、AGO组中Th17细胞比例较C组升高(P<0.05),且AGO组升高最明显。RORγt在GD组、NGO组、AGO组中升高(P<0.05),其中AGO组升高最明显。TGF-β、IL-35在GD组、NGO组、AGO组中表达下降(P<0.05),IL-10在3组中表达升高(P<0.05);GD组、NGO组、AGO组中IL-17A、IL-23、IL-6蛋白较C组比较升高(P<0.05),且AGO组中升高更明显(P<0.05)。结论 Treg细胞免疫抑制功能的降低可能是促使Graves眼病发生的一个重要因素。Th17细胞可能参与介导Graves眼病的发生与发展,Th17细胞及相关细胞因子可能是评估GO患者病情活动性的新指标。Treg/Th17平衡的破坏可能参与Graves眼病的发病。
Objective To explore the role of CD4＋CD25＋Foxp3＋Treg/CD4＋IL-17A＋Th17 cells and the related cytokines in Graves＇ ophthalmopathy. Methods Based on clinical activity scores （CAS）, we divided patients with untreated Graves＇ ophthal-mopathy into active group （AGO group with CAS≥3 （15 cases） and non-active group （NGO group） with CAS〈3 （15 cases）, with another 15 patients with untreated Graves＇ disease free of eye symptoms （GD group） and 15 normal subjects as controls. Peripheral venous blood Treg/Th17 cell ratio was determined using flow cytometry. RT-PCR was used to detect the mRNA expression levels of Treg-specific transcription factor Foxp3 and Th17-specific transcription factor RORγt. Enzyme-linked immunosorbent assay （ELISA） was used to detect the serum levels of Th17 cell-related cytokines （IL-17A, IL-23, and IL-6） and Treg-related cytokines （TGF-β, IL-10, and IL-35）. Results Compared with the normal subjects, the patients in GD, NGO, AGO groups all showed significantly increased Th17 cell count （P〈0.05）, which was the highest in AGO group. RT-PCR results revealed significantly increased RORγt in GD, NGO, and AGO groups, also the highest in AGO group. Serum IL-17A, IL-23, and IL-6 levels all showed significant increments in GD, NGO, and AGO groups （P〈0.05）, especially in AGO group. Among the Treg-related cytokines, TGF-βand IL-35 levels decreased （P〈0.05） but IL-10 increased significantly （P〈0.05） in GD, NGO, AGO groups. Conclusion Decreased immunosuppressive capacity of Treg cells can be an important factor in the pathogenesis of Graves＇ ophthalmopathy. Th17 cells may also participate in the occurrence and progression of Graves＇ ophthalmopathy and can serve along with related cytokines as novel indicators of the disease activity. Impaired Treg/Th17 balance may importantly contribute to the occurrence of Graves＇ ophthalmopathy.
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2014年第12期1809-1813,共5页
Journal of Southern Medical University
基金
广东省自然科学基金(S2013010016045)
