Urinary haptoglobin, alpha‐1 anti‐chymotrypsin and retinol binding protein identified by proteomics as potential biomarkers for lupus nephritis

A. Aggarwal, R. Gupta, V. Negi, L. Rajasekhar, R. Misra, P. Singh, V. Chaturvedi, S. Sinha
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引用次数: 32

Abstract

The study was aimed at identification by proteomics and validation by enzyme‐linked immunosorbent assay (ELISA) of potential urinary biomarkers for lupus nephritis. Study subjects comprised 88 systemic lupus erythematosus (SLE) patients and 60 controls (rheumatoid arthritis, diabetes mellitus and healthy individuals). Based on the SLE disease activity index (SLEDAI), patients were classified as active renal (AR), active non‐renal (ANR) or inactive disease (ID). Urinary proteins from a group of patients with AR or ID were resolved by two‐dimensional gel electrophoresis and identified by matrix‐assisted laser desorption ionization–time of flight–mass spectrometry (MALDI‐TOF‐MS/MS). The selected biomarkers were validated by ELISA using samples from all patients and controls. AR patients were followed‐up for 12 months after start of therapy. Three urinary proteins, alpha‐1 anti‐chymotrypsin (ACT), haptoglobin (HAP) and retinol binding protein (RBP), were detected in patients with AR and not ID. Upon validation, ACT levels were higher in AR patients than the other groups (P < 0·001) and showed good correlation with renal SLEDAI (r = 0·577, P < 0·001) as well as SLEDAI (r = 0·461, P < 0·001). Similarly, HAP levels were > 10‐fold higher in AR than other groups (P < 0·001) and correlated well with renal SLEDAI (r = 0·594, P < 0·001) and SLEDAI (r = 0·371, P < 0·01). RBP levels were also higher in AR patients than in other groups (P < 0·05), except diabetes, and showed moderate correlation with renal SLEDAI (r = 0·284, P < 0·008) and SLEDAI (r = 0·316, P < 0·003). Upon follow‐up with treatment, levels of all three proteins declined at 6 and 12 months (P < 0·01). Multiple logistic regression identified ACT as the best marker to differentiate AR from ANR. Urinary HAP, ACT and RBP are potential biomarkers for lupus nephritis activity.
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尿触珠蛋白、α - 1抗凝乳胰蛋白酶和视黄醇结合蛋白被蛋白质组学鉴定为狼疮性肾炎的潜在生物标志物
该研究旨在通过蛋白质组学鉴定和酶联免疫吸附试验(ELISA)验证狼疮肾炎的潜在尿液生物标志物。研究对象包括88例系统性红斑狼疮(SLE)患者和60例对照(类风湿关节炎、糖尿病和健康个体)。根据SLE疾病活动性指数(SLEDAI),将患者分为活动性肾脏(AR)、活动性非肾脏(ANR)和非活动性疾病(ID)。采用二维凝胶电泳和基质辅助激光解吸电离飞行时间质谱法(MALDI - TOF - MS/MS)对一组AR或ID患者的尿蛋白进行了鉴定。选择的生物标志物通过ELISA对所有患者和对照组的样本进行验证。AR患者在治疗开始后随访12个月。在AR患者而非ID患者中检测到三种尿蛋白,α - 1抗凝乳胰蛋白酶(ACT)、触珠蛋白(HAP)和视黄醇结合蛋白(RBP)。经验证,AR患者的ACT水平高于其他组(P < 0.001),且与肾脏SLEDAI (r = 0.594, P < 0.001)和SLEDAI (r = 0.371, P < 0.01)有良好的相关性。除糖尿病外,AR患者RBP水平均高于其他组(P < 0.05),且与肾脏SLEDAI (r = 0.284, P < 0.008)和SLEDAI (r = 0.316, P < 0.003)呈中度相关。治疗后随访6个月和12个月,三种蛋白水平均下降(P < 0.01)。多元逻辑回归发现ACT是区分AR和ANR的最佳标记。尿HAP、ACT和RBP是红斑狼疮肾炎活动性的潜在生物标志物。
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