【作者】 邢倩；

【导师】 王斌；

【作者基本信息】 青岛大学 ， 病原生物学， 2011， 博士

【摘要】 目的研究系统性红斑狼疮(SLE)患者外周血中T辅助细胞17(Th17细胞)、调节性T细胞(Treg)亚群的比例,外周血CD4+ T细胞中CD25和FoxP3的表达,Th17、Treg细胞特异性转录因子RORγt、FoxP3基因表达水平,检测血清中相关细胞因子白细胞介素-17(IL-17)、干扰素-γ(IFN-γ)、白细胞介素-4(IL-4)、血清和尿中转化生长因子-β1(TGF-β1)水平的变化,揭示Th17、Treg细胞及相关细胞因子在狼疮性肾炎发病中的免疫机制作用。方法选取60例系统性红斑狼疮患者和28例相匹配的健康体检者作为对照,采用流式细胞仪检测所有受试者外周血单个核细胞(PBMC)中的Th17细胞和CD4+CD25+、CD4+CD25high、CD4+CD25+FoxP3+T细胞的百分率,检测FoxP3在CD4+CD25high、CD4+CD25-中的表达。逆转录聚合酶链反应(RT-PCR)检测PBMC中RORγtmRNA、FoxP3 mRNA的表达水平,酶联免疫吸附测定法(ELISA法)检测血清中IL-17、IL-2、IL-4、血清和尿中TGF-β1水平,并对其与SLEDAI、SDI.24小时尿蛋白量相关性进行研究。结果1.狼疮性肾炎组患者外周血中Th17细胞百分率与狼疮无肾炎组、健康对照组相比显著升高(P<0.05,P<0.01)；CD4+T细胞中的CD4+CD25+、CD4+CD25high和CD4+CD25+FoxP3+T细胞百分率显著降低(P<0.05, P<0.01); FoxP3在CD4+CD25highT细胞中的表达率及平均免疫荧光强度(MFI)较狼疮无肾炎组及健康对照组患者显著降低(P<0.05,P<0.01)；FoxP3在CD4+CD25-T细胞中的表达率较健康对照组显著增高(P<0.01)。2.狼疮性肾炎组患者外周血中RORγt mRNA表达水平与健康对照组、狼疮无肾炎组相比显著升高(P<0.05)；狼疮性肾炎组FoxP3 mRNA表达水平与健康对照组相比显著降低(P<0.05)。3.狼疮性肾炎组患者血清IL-17、IFN-γ、IL-4水平均显著高于狼疮无肾炎组和健康对照组(P<0.001),血清TGF-β1水平显著低于健康对照组水平(P<0.001),而尿TGF-β1水平显著高于狼疮无肾炎组和健康对照组(P<0.001),尿TGF-β1表达水平与24小时尿蛋白量成正相关(P<0.01)。结论狼疮性肾炎患者外周血中Th17细胞比例显著升高,Treg细胞比例显著降低,转录因子RORγt mRNA表达水平升高而FoxP3 mRNA表达水平降低,外周血中转录因子失衡导致Th17/Treg免疫偏移,体内的免疫抑制效应减弱而免疫炎性效应增强,继而细胞因子网络失衡,免疫耐受缺失,促炎细胞因子升高促进狼疮性肾炎的发生、发展。更多还原

【Abstract】 ObjectiveTo investigate the variations of T-helper 17 (Th17) and regulatory T (Treg) cells, assess the expression of FoxP3 and CD25 on CD4+ regulatory T (Treg) cells and the expression of RORyt gene and FoxP3 gene, measure the level of Th cells related cytokines and their clinical significance in patients with systemic lupus erythematosus (SLE), evaluate the role of Th17 and Treg cells in the pathogenesis of patients with lupus nephritis (LN).MethodsA total of 60 systemic lupus erythematosus patients and 28 healthy controls were enrolled. The frequency of Th17 cells and Treg cells in peripheral blood mononuclear cells (PBMC) was evaluated by flow cytometry (FACS). The expression of CD25 and FoxP3 by CD4+ T cells was analyzed by flow cytometric analysis. RT-PCR was used to measure the expression of RORyt gene and FoxP3 gene. The concentrations of serum IL-17, IL-4, IFN-y, TGF-β1 and urinary TGF-β1 were measured by enzyme-linked immunosorbent assay (ELISA).Results1. A significant increase in the frequency of Th17 cells and a significant decrease in the frequency of CD4+CD25+、CD4+CD25high and CD4+CD25+FoxP3+ T cells in peripheral blood of LN patients compared with SLE group and healthy controls (P< 0.05, P<0.01; respectively). LN patients had a lower percentage and expression of FoxP3 in CD4+CD25high T cells than SLE patients without nephritis (P< 0.05).2. The expression of RORyt mRNA levels in LN patients exhibited a significant increase compared with patients without nephritis and healthy controls (P< 0.05). The expression of FoxP3 mRNA levels in LN patients exhibited a significant decrease compared with healthy controls (P<0.05). 3. The concentration of serum IL-17, IL-4, IFN-y was found increased in LN patients compared with that from healthy controls (P< 0.001), while serum TGF-β1 was found decreased in SLE patients compared with that from healthy controls (P< 0.001). Urinary TGF-β1 levels were significantly higher in SLE patients with nephritis in comparison to those without nephritis and healthy controls (P<0.001), urinary TGF-β1 levels correlated positively with 24h urine protein (P<0.01).ConclusionsThe significantly elevated Th17 cells were accompanied by FoxP3+Treg cells decrease in lupus nephritis, suggesting that Th17/Treg functional imbalance may be involved in the pathogenesis of renal damage in SLE patients.更多还原