抗体芯片在慢性肾脏病患者尿中多种细胞因子同步检测中的初步应用

张露;刘必成;王艳丽;张晓良;吕林莉;刘殿阁

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中华肾脏病杂志 ›› 2006, Vol. 22 ›› Issue (2) : 119-123.
新技术与方法

抗体芯片在慢性肾脏病患者尿中多种细胞因子同步检测中的初步应用

  • 张露; 刘必成; 王艳丽; 张晓良; 吕林莉; 刘殿阁
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Application of antibody array technology in analysis of urinary cytokine profiles in patients with chronic kidney disease

  • ZHANG Lu; LIU Bi-cheng; WANG Yan-li;ZHANG Xiao-liang;
    LV Lin-li; LIU Dian-Ge
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摘要

目的 应用抗体芯片技术检测慢性肾脏病(CKD)患者尿液中细胞因子的水平,并探讨其临床意义。方法 研究对象共10例,7例为CKD患者,全部符合 K/DOQI指南中CKD的诊断标准,并且有肾脏病理诊断。依据GFR水平以及CKD分期,将7例患者分为两组:Ⅰ组:GFR≥80 ml•min-1•(1.73 m2)-1(CKD1~2期)共4例; Ⅱ组:GFR≤40 ml•min-1•(1.73 m2)-1(CKD3~5期)共3例。另选取3例性别、年龄相匹配的健康人作为正常对照。应用Raybiotech人类细胞因子抗体芯片,同时检测各组尿液中20种细胞因子水平的变化。结果 与正常对照组相比,CKD患者共有15种因子的水平发生了显著变化。单核细胞趋化蛋白(MCP)-1、RENTES、金属蛋白酶组织抑制物(TIMP)-1、肿瘤坏死因子(TNF)-α、血管内皮生长因子(VEGF)、E-选择素(seletin)、Fas、细胞间黏附分子(ICAM)-1、白细胞介素(IL)-2、基质金属蛋白酶(MMP)-2、转化生长因子(TGF)-β和血小板源生长因子(PDGF)-BB的水平同正常对照组相比升高了2~5倍,其中尿MCP-1, RANTES, TIMP-1, TNF-α和 VEGF的水平随着GFR的下降而进一步升高。VCAM-1 和PDGF-BB的水平同正常对照组相比有所下降。在芯片所包含的20种细胞因子中,MMP-9的水平变化尤其显著,同正常对照组相比,CKDⅠ组是正常对照组的492.8倍,CKDⅡ组是正常对照组的198.7倍。结论 首次应用抗体芯片技术,证实CKD患者尿液中细胞因子表达水平同正常对照组有明显差异,并且与疾病所处阶段有一定的关系。

Abstract

Objective To apply a novel proteomic technology to analyze urinary cytokine profiles in patients with CKD. Methods Ten subjects including 7 CKD patients and 3 normal controls were enrolled in this study. Differential excretion of urinary cytokines was determined by human cytokines antibody array (Raybiotech Norcross GA). As compared to controls, a two-fold change in spot intensity was considered significant. Results A total of 15 cytokines varied significantly in urinary samples from patients with CKD, comparing with controls. It was shown that MCP-1, RANTES, TIMP-1,TNF-α, VEGF, E-selectin, Fas, ICAM-1, IL-2, MMP-2, TGF-β increased by two to five folds in CKD patients with normal renal function, while the excretion of MCP-1, RANTES, TIMP-1, TNF-α and VEGF was further increased with the declining of renal function. A decreasing excretion of urinary VCAM-1 and PDGF was found in patients with renal failure. Impressively, the urinary MMP-9 excretion was 492 folds increase in CKD patients with normal renal function and 198 folds increase in renal failure patients as compared to normal controls. Conclusion A significant change of urinary cytokine profiles in patients with CKD is firstly demonstrated as compared to normal controls by using a novel antibody array technology, which may provide important information regarding to the role of cytokines in the progression of CKD.

关键词

蛋白质阵列分析 / 肾功能衰竭 / 慢性 / 尿 / 细胞因子类

Key words

Protein array analysis / Kidney failure / chronic / Urine / Cytokines

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张露;刘必成;王艳丽;张晓良;吕林莉;刘殿阁. 抗体芯片在慢性肾脏病患者尿中多种细胞因子同步检测中的初步应用[J]. 中华肾脏病杂志, 2006, 22(2): 119-123.
ZHANG Lu;LIU Bi-cheng;WANG Yan-li;ZHANG Xiao-liang;LV Lin-li;LIU Dian-Ge. Application of antibody array technology in analysis of urinary cytokine profiles in patients with chronic kidney disease[J]. Chinese Journal of Nephrology, 2006, 22(2): 119-123.
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