IL-1ra抑制IL-1α诱导的眼眶成纤维细胞表达ICAM-1

IL-lra Inhibits IL-la-induced ICAM-1 Expression on Cultured Orbital Fibroblasts

【目的】明确IL-1ra是否对IL-1α诱导的眼眶成纤维细胞表达ICAM-1有抑制作用及生物学意义,探讨表达ICAM-1在IL-1α诱导的外周血单个核细胞与眼眶成纤维细胞黏附中所起的作用。【方法】以IL-1α或/和IL-1ra处理培养的Graves’眼病(GO)患者和正常人眼眶成纤维细胞。用半定量RT-PCR法检测IL-1ra对IL-1α诱导的眼眶成纤维细胞表达ICAM-1 mRNA的影响;用Western blot法和细胞免疫化学法检测IL-1ra对IL-1α诱导的眼眶成纤维细胞表达ICAM-1蛋白的影响;用荧光显微镜检测预先标记的外周血单个核细胞(PBMC)与培养的眼眶成纤维细胞的黏附能力。用抗ICAM-1中和抗体来显示ICAM-1在IL-1α诱导的黏附过程中所起的作用。【结果】IL-1ra抑制IL-1α诱导的GO患者和正常人眼眶成纤维细胞表达ICAM-1 mRNA和蛋白呈剂量依赖性;IL-1ra抑制IL-1α诱导的PBMC与眼眶成纤维细胞的黏附呈浓度及时间依赖性。ICAM-1 mRNA和蛋白被抑制的方式与眼眶成纤维细胞受IL-1ra处理后与PBMC黏附的减弱明显平行。而且,抗ICAM-1单克隆抗体抑制PBMC黏附到眼眶成纤维细胞呈浓度依赖性。【结论】IL-1ra能够在mRNA和蛋白水平抑制IL-1α诱导的眼眶成纤维细胞表达ICAM-1,IL-1ra抑制IL-1α诱导的眼眶成纤维细胞表达ICAM-1可能在减弱IL-1α诱导的PBMC黏附到成?

[ Objective ] To identify whether interleukin-1 receptor antagonist (IL-1 ra) inhibits inter-leukin-1 a(IL-1*)-induced intercellular adhesion molecule-1 (ICAM-1) expression on cultured orbital fibroblasts and its biological role. To investigate the function of ICAM-1 in IL-la-induced adhesion of peripheral blood mononuclear cells (PBMC) with orbital fibroblasts. [Methods] Cultured orbital fibroblasts from patients with Graves' ophthalmopathy(GO) and controls were challenged with IL-1α or/and IL-1ra. Semi-quantitative analysis of RT-PCR was used to examine the influence of IL-1ra on IL-lα-induced ICAM-1 mRNA expression on cultured orbital fibroblasts. Western blot and immunocyto-chemical staining were used to show the changes of ICAM-1 protein in this treatment. Fluoromi-croscopy was used to show the adhesion capacity between the labelled PBMC and orbital fibroblasts. ICAM-1 monoclonal antibody was used to demonstrate the roles of ICAM-1 in the IL-1α-induced adhesion. [Results] IL-1ra inhibits IL-1α-induced ICAM-1 mRNA and protein expression on cultured orbital fibroblasts both from GO patients and controls in dose-dependent manner. IL-lra inhibits IL-la-induced adhesion between PBMC and orbital fibroblasts both in dose-dependent and time-dependent manners. The inhibition manner of ICAM-1 mRNA and protein expression on cultured orbital fibroblasts obviously paralleled with the inhibition manner of orbital fibroblasts adhesion with PBMC after IL-1ra treatment. Moreover, ICAM-1 monoclonal antibody inhibits the adhesion of PBMC and fibroblasts in concentration-dependent manner. [Conclusions] IL-lra inhibits IL-1α-induced I-CAM-1 expression on cultured orbital fibroblasts both on mRNA and protein levels, and it inhibits the adhesion of PBMC to fibroblasts. The inhibition of IL-la-induced ICAM-1 expression by IL-lra may play an important role in the inhibition of IL-la-induced PBMC adhesion with fibroblasts. IL-lra may be useful in the prevention or treatment of G0.