Wang Zhiyi, Shi Yujia, Yang Yachen, Gong Bangdong, Xie Jianmin
Department of Rheumatology, The Second Affiliated Hospital of Nanjing Medical University, Nanjing, China.
Department of Rheumatology, Tongji Hospital of Tongji University, Shanghai, China.
Front Immunol. 2025 May 19;16:1577590. doi: 10.3389/fimmu.2025.1577590. eCollection 2025.
Loss of gut barrier integrity has been observed in rheumatoid arthritis (RA). While systemic inflammation in RA has been extensively investigated, intestinal-specific inflammatory processes remain poorly understood. This study is designed to identify a novel biomarker panel combining fecal cytokine profiles with gut barrier biomarkers to discriminate RA patients with varying disease progression.
Feces (Fc) and plasma (Pl) were obtained from 62 Naive RA patients (NA), 47 remission RA patients (RE), 28 difficult-to-treat RA patients(D2T), and 70 healthy controls (HC). A panel of 12 cytokines and gut barrier markers, including intestinal Fatty-Acid-Binding Protein-2 (FABP2), zonulin, Hypoxia-Inducible Factor-2α (HIF-2α), D-lactate, LBP and fecal calprotectin (FCAL), was quantified by ELISA. Statistical integration with clinical parameters was performed using univariate and multivariate approaches.
NA and D2T patients demonstrated marked elevations in fecal pro-inflammatory cytokines compared to RE and HC groups, including IL-6, Granulocyte-Macrophage Colony-Stimulating Factor (GM-CSF), IL-1 beta (IL-1β), Interferon-gamma (INF-γ), IL-23, Tumor Necrosis Factor-Alpha (TNF-α), IL-21, IL-17A/F, and IL-22. Fecal zonulin and plasma HIF-2α were significantly elevated in both NA and D2T groups, whereas fecal D-lactate showed a pronounced decrease in the NA and D2T groups. These biomarkers demonstrated the strongest correlation with disease severity indices. Receiver operating characteristic (ROC) analysis revealed that fecal FABP2, zonulin and D-lactate exhibited superior discriminative capacity between the NA and RE groups. whereas fecal zonulin showed remarkable diagnostic potential for distinguishing NA from D2T groups compared to plasma counterparts. The discriminant scores (DS) model incorporating fecal zonulin and plasma HIF-2α demonstrated superior discriminatory performance between the D2T and NA groups compared to the model utilizing the top five plasma parameters.
Our fecal profiling methodology provides novel insights into the gut mucosal cytokine microenvironment during RA progression. The dissociation between fecal and plasma inflammatory profiles underscores the critical importance of localized gut immune monitoring in RA management.
在类风湿关节炎（RA）中已观察到肠道屏障完整性受损。虽然RA中的全身炎症已得到广泛研究，但肠道特异性炎症过程仍知之甚少。本研究旨在确定一种新的生物标志物组合，将粪便细胞因子谱与肠道屏障生物标志物相结合，以区分疾病进展不同的RA患者。
从62例初发RA患者（NA）、47例缓解期RA患者（RE）、28例难治性RA患者（D2T）和70例健康对照（HC）中获取粪便（Fc）和血浆（Pl）。通过酶联免疫吸附测定（ELISA）对包括肠道脂肪酸结合蛋白-2（FABP2）、闭合蛋白、缺氧诱导因子-2α（HIF-2α）、D-乳酸、脂多糖结合蛋白（LBP）和粪便钙卫蛋白（FCAL）在内的12种细胞因子和肠道屏障标志物进行定量分析。使用单变量和多变量方法对临床参数进行统计整合。
与RE组和HC组相比，NA组和D2T组患者粪便中的促炎细胞因子显著升高，包括白细胞介素-6（IL-6）、粒细胞-巨噬细胞集落刺激因子（GM-CSF）、白细胞介素-1β（IL-1β）、干扰素-γ（INF-γ）、白细胞介素-23、肿瘤坏死因子-α（TNF-α）、白细胞介素-21、白细胞介素-17A/F和白细胞介素-22。NA组和D2T组的粪便闭合蛋白和血浆HIF-2α均显著升高，而NA组和D2T组的粪便D-乳酸则显著降低。这些生物标志物与疾病严重程度指数的相关性最强。受试者工作特征（ROC）分析显示，粪便FABP2、闭合蛋白和D-乳酸在NA组和RE组之间具有卓越的鉴别能力。与血浆标志物相比，粪便闭合蛋白在区分NA组和D2T组方面显示出显著的诊断潜力。与使用前五个血浆参数的模型相比，包含粪便闭合蛋白和血浆HIF-2α的判别分数（DS）模型在D2T组和NA组之间表现出更优的鉴别性能。
我们的粪便分析方法为RA进展过程中的肠道黏膜细胞因子微环境提供了新的见解。粪便和血浆炎症谱之间的差异凸显了局部肠道免疫监测在RA管理中的至关重要性。
