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目的研究特发性膜性肾病(IMN)患者外周血和肾组织中调节性T细胞(Treg)和辅助性T细胞17(Th17)的比例变化,探讨Treg和Th17比例失衡参与IMN发病的可能机制。方法选取于上海交通大学医学院附属新华医院就诊经肾活检确诊为IMN的患者(IMN组,n=30)、健康志愿者(健康对照组,n=20)及行肾癌手术者(肾癌对照组,n=10)为研究对象。采用流式细胞仪测定IMN组和健康对照组外周血单核细胞(PBMC)中Treg和Th17的表达;实时聚合酶链反应(PCR)法检测PBMC中叉状头/翅膀状螺旋转录因子3(Foxp3)和维甲酸相关孤核受体(RORγt)的表达水平;采用免疫组化法检测IMN组和肾癌对照组肾组织中IL-17的表达。结果流式细胞仪检测结果显示,与健康对照组相比,IMN组PBMC中CD4~+Foxp3~+Treg表达水平降低(P<0.05),Th17表达水平升高(P<0.05),Treg/Th17比值显著降低(P<0.01)。PCR检测结果显示,与健康对照组相比,IMN组PBMC中Foxp3 mRNA的表达水平明显下调(P<0.01),RORγt mRNA水平明显上调(P<0.01)。免疫组化结果显示,IMN组患者肾组织中IL-17的表达较肾癌对照组患者显著升高(P<0.05),主要体现于肾小管处。相关性分析显示,IMN组患者外周血Treg/Th17比值与24 h尿蛋白定量水平呈明显负相关(r=-0.537,P<0.05),与血清白蛋白水平呈明显正相关(r=0.669,P<0.05)。结论 IMN患者外周血中存在Treg表达水平降低和Th17表达水平升高的比例失衡,而Treg/Th17比例失衡与IMN的发病机制有关。
Objective To study the proportion of regulatory T cells (Tregs) and helper T cells (Th17) in peripheral blood and kidney of patients with idiopathic membranous nephropathy (IMN), and to explore the possible mechanism of imbalance of Treg and Th17 in the pathogenesis of IMN . Methods Patients (IMN group, n = 30) diagnosed as IMN by renal biopsy at Xinhua Hospital Affiliated to Shanghai Jiaotong University School of Medicine were selected, healthy volunteers (n = 20) and renal cell carcinoma patients Control group, n = 10) as the research object. Flow cytometry was used to detect the expression of Treg and Th17 in peripheral blood mononuclear cells (PBMCs) of IMN group and healthy control group. Real-time polymerase chain reaction (PCR) was used to detect the expression of forkhead / wing helical transcription factor 3 Foxp3) and retinoic acid-related orphan nuclear receptor (RORγt) were detected by immunohistochemistry. The expression of IL-17 in renal tissues of IMN group and control group was detected by immunohistochemistry. Results The results of flow cytometry showed that the expression of CD4 ~ + Foxp3 ~ + Treg in PBMC of IMN group was significantly lower than that in healthy control group (P <0.05), while the expression of Th17 was increased (P <0.05) Ratio was significantly lower (P <0.01). The results of PCR showed that the expression of Foxp3 mRNA in PBMC of IMN group was significantly lower than that of healthy control group (P <0.01), and the mRNA level of RORγt was significantly increased (P <0.01). The results of immunohistochemistry showed that the expression of IL-17 in renal tissue of IMN group was significantly higher than that of the control group (P <0.05), mainly in the renal tubule. Correlation analysis showed that the Treg / Th17 ratio in peripheral blood was negatively correlated with 24 h urinary protein level (r = -0.537, P <0.05), and positively correlated with serum albumin level (r = 0.669, P <0.05). Conclusion There is an imbalance between the decrease of Treg expression and the increase of Th17 expression in peripheral blood of patients with IMN. The imbalance of Treg / Th17 is related to the pathogenesis of IMN.