Inhibition of matrix metalloproteinase-3 synthesis in human conjunctival fibroblasts by interleukin-4 or interleukin-13
Inhibition of matrix metalloproteinase-3 synthesis in human conjunctival fibroblasts by interleukin-4 or interleukin-13
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DOI：
10.1167/iovs.05-1261
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发表时间：
2006-07-01
影响因子：
4.4
通讯作者：
Nishida, T
中科院分区：
文献类型：
--
作者：
Fukuda, K;Fujitsu, Y;Nishida, T
目的.结膜纤维增生性病变称为巨大乳头是春季角结膜炎（VKC）的特征。在VKC患者的巨乳头和泪液中，辅助性T细胞2（Th 2）和细胞因子的丰度增加，Th 2细胞因子白介素（IL）-4和IL-13各自刺激结膜成纤维细胞产生细胞外基质（ECM）蛋白。通过测定Th 2细胞因子对结膜成纤维细胞产生基质金属蛋白酶（MMP）-3（ECM降解的关键酶）的影响，进一步研究Th 2细胞因子在巨乳头发育中的作用。通过酶联免疫吸附试验测定人结膜成纤维细胞释放到培养基中的MMP-3的量，并通过逆转录和实时聚合酶链反应分析定量MMP-3 mRNA的细胞内丰度。通过免疫印迹和免疫荧光分析评估转录因子NF-κ B和AP-1的信号传导。在测试的Th 2细胞因子中，仅IL-4和IL-13抑制结膜成纤维细胞的MMP-3的基础和IL-1 β诱导的释放。IL-4和IL-13的这些作用被IL-4受体复合物的中和抗体抑制。IL-4和IL-13也分别降低了这些细胞中MMP-3 mRNA的基础丰度，并抑制了IL-1 β诱导的MMP-3 mRNA上调。IL-4和IL-13均不影响IL-1 β诱导的NF-κ B或AP-1组分的活化。IL-4和IL-13各自抑制人结膜成纤维细胞中的MMP-3合成，表明这些Th 2细胞因子可能通过阻止该酶介导的基质降解而导致ECM在巨乳头中的过度沉积。
PURPOSE. Fibroproliferative lesions of the conjunctiva known as giant papillae are a characteristic of vernal keratoconjunctivitis (VKC). The abundance of T helper 2 (Th2) cells and cytokines is increased in the giant papillae and tear fluid of individuals with VKC, and the Th2 cytokines interleukin (IL)-4 and IL-13 each stimulate the production of extracellular matrix (ECM) proteins by conjunctival fibroblasts. The role of Th2 cytokines in the development of giant papillae was further examined by determination of the effects of these molecules on the production by conjunctival fibroblasts of matrix metalloproteinase (MMP)-3, a key enzyme in ECM degradation.METHODS. The amount of MMP-3 released into the culture medium by human conjunctival fibroblasts was determined by enzyme-linked immunosorbent assay, and the intracellular abundance of MMP-3 mRNA was quantitated by reverse transcription and real-time polymerase chain reaction analysis. Signaling by the transcription factors NF-kappa B and AP-1 was evaluated by immunoblot and immunofluorescence analyses.RESULTS. Of the Th2 cytokines tested, only IL-4 and -13 inhibited both the basal and IL-1 beta-induced release of MMP-3 by conjunctival fibroblasts. These effects of IL-4 and -13 were inhibited by neutralizing antibodies to the IL-4 receptor complex. IL-4 and -13 also each reduced the basal abundance, as well as inhibited the IL-1 beta-induced upregulation, of MMP-3 mRNA in these cells. Neither IL-4 nor -13 affected the IL-1 beta-induced activation of NF-kappa B or the AP-1 component c-Jun.CONCLUSIONS. IL-4 and -13 each inhibit MMP-3 synthesis in human conjunctival fibroblasts, suggesting that these Th2 cytokines may contribute to the excessive deposition of ECM in giant papillae by preventing matrix degradation mediated by this enzyme.