摘要
目的初步探讨瘦素对RA患者外周血调节性T细胞增殖的影响。方法分离培养6例RA患者PBMCs,分别用抗CD3(3 μg/ml)/抗CD28(2 μg/ml)、抗CD3(3 μg/ml)/抗CD28(2 μg/ml)加瘦素(100 ng/ml)、抗CD3(3 μg/ml)/抗CD28(2 μg/ml)以及瘦素(100 ng/ml)加IL-6R拮抗剂(0.5 μg/ml)刺激72 h。收集悬浮细胞,流式细胞术检测调节性T细胞百分率;ELISA检测细胞培养上清中IL-6、TGF-β水平。采用Wilcoxon秩和检验进行统计学分析。结果①与对照组比较,瘦素刺激72 h后调节性T细胞比例下降[13.6(11.6,14.8)%和9.8(7.0,10.0)%,Z=-2.201,P〈0.05];IL-6、TGF-β水平均上升[548.9(508.6,608.4)ng/L和631.8(538.4,672.6)ng/L,Z=-1.992,P〈0.05;1175.0(1 045.4,1 373.1)ng/L和1 580.2(1 315.3,1 906.5)ng/L,Z=-2.201,P〈0.05];②加入IL-6R拮抗剂后,瘦素刺激PBMCs分泌IL-6水平下降[631.8(538.4,672.6)ng/L和522.7(339.3,593.3)ng/L,Z=-2.201,P〈0.05];调节性T细胞比例上升[9.8(7.0,10.0)%和12.3(9.7,13.8)%,Z=-1.997,P〈0.05]。结论瘦素可能通过上调IL-6,进而对外周血调节性T细胞的增殖起到负调控作用。
ObjectiveTo investigate the influence of Leptin on the proliferation of peripheral blood regulatory T cells in patients with rheumatoid arthritis (RA).MethodsPeripheral blood mononuclear cells (PBMCs) were isolated from 6 RA patients. RA PBMCs were stimulated with anti-CD3 (3 μg/ml)/anti-CD28 (2 μg/ml) antibody for 72 h, anti-CD3 (3 μg/ml)/anti-CD28 (2 μg/ml) antibody and Leptin (100 ng/ml) for 72 h, anti-CD3 (3 μg/ml)/anti-CD28 (2 μg/ml) antibody and Leptin (100 ng/ml) and interleukin (IL)-6 receptor antagonist (0.5 μg/ml) for 72 h. Regulatory T cells (Treg) were detected by flow cetometry. IL-6 level and TGF-β levels in the supernatants of RA PBMCs culture were detected by Enzyme linked immunosorbent assay(ELISA). Statistical analysis between the two groups were performed by Wilcoxon Signed Rank Test.Results① Our results suggested that compared to the control group, RA PBMCs were stimulated by leptin for 72 h had lower percentage of Treg[13.6(11.6, 14.8)% vs 9.8(7.0, 10.0)%, Z=-2.201, P〈0.05]; Compared to the control group, the level of IL-6 and TGF-β were elevated when RA PBMCs treated with leptin for 72 h [548.9(508.6, 608.4) ng/L vs 631.8 (538.4, 672.6) ng/L, Z=-1.992, P〈0.05; 1175.0 (1 045.4, 1 373.1) ng/L vs 1 580.2 (1 315.3, 1 906.5) ng/L, Z=-2.201, P〈0.05]; ② The percentage of Peripheral Treg was increased [9.8(7.0,10.0)% vs 12.3 (9.7, 13.8)%, Z=-1.997, P〈0.05] and the level of IL-6 was decreased [631.8(538.4, 672.6) ng/L vs 522.7(339.3, 593.3) ng/L, Z=-2.201, P〈0.05] when RA PBMCs was stimulated with IL-6 receptor antagonist and anti-CD3/anti-CD28 antibody and Leptin for 72 h.ConclusionThe study has shown that Leptin can down-regulate the percentage of RA peripheral blood Treg via increasing IL-6 secretion.
作者
叶芸杉
李寒
欧阳寻丽
袁启航
王冰
张彦
Ye Yunshan;Li Hart;Ouyang Xunli;Yuan Qihang;Wang Bing;Zhang Yan(Department of Immunology,College of Basic Medicine Science,Dalian Medical University,Liaoning 116044,China)
出处
《中华风湿病学杂志》
CAS
CSCD
北大核心
2018年第10期660-663,共4页
Chinese Journal of Rheumatology
基金
国家自然科学基金(81671606
81373214)
辽宁省高等学校基本科研项目(LQ2017004)
大连医科大学转化医学专项基金(2015010)