Dexamethasone inhibition of IL-1-induced collagen degradation by corneal fibroblasts in three-dimensional culture.
Dexamethasone inhibition of IL-1-induced collagen degradation by corneal fibroblasts in three-dimensional culture.
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DOI：
10.1167/iovs.04-0051
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发表时间：
2004-09
影响因子：
4.4
通讯作者：
Ying Lu;K. Fukuda;Yang Liu;N. Kumagai;T. Nishida
中科院分区：
文献类型：
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作者：
Ying Lu;K. Fukuda;Yang Liu;N. Kumagai;T. Nishida
目的：糖皮质激素调节炎症细胞的功能。本研究的目的是研究地塞米松对角膜成纤维细胞胶原降解的影响，胶原降解是角膜溃疡的潜在原因。方法采用I型胶原三维凝胶培养兔角膜成纤维细胞，不添加或不添加il -1 β或地塞米松。通过测定培养上清酸热水解产生的羟脯氨酸的量来测定胶原降解的程度。采用免疫印迹分析、明胶酶谱法、逆转录和实时聚合酶链反应检测基质金属蛋白酶（MMPs）和金属蛋白酶组织抑制剂（TIMPs）的表达。采用免疫印迹法检测角膜成纤维细胞中丝裂原活化蛋白激酶（MAPKs）的磷酸化水平。结果地塞米松抑制il -1 β诱导的角膜成纤维细胞降解胶原呈剂量依赖性。地塞米松抑制了MMPs的合成和激活，抑制了TIMPs的表达，也抑制了培养上清中纤溶酶的活性。地塞米松还能抑制il -1 β诱导的MAPKs细胞外信号调节激酶（ERK）和c-Jun n -末端激酶（JNK）的磷酸化，但对p38没有抑制作用。结论地塞米松对角膜成纤维细胞的MMP-TIMP系统有多重作用，从而抑制il -1诱导的胶原降解。抑制il -1 β诱导的ERK和JNK的激活可能有助于地塞米松的这些作用。
PURPOSE Corticosteroids regulate the functions of inflammatory cells. The purpose of the present study was to investigate the effect of dexamethasone on collagen degradation by corneal fibroblasts, an underlying cause of corneal ulceration. METHODS Rabbit corneal fibroblasts were cultured in three-dimensional gels of type I collagen and in the absence or presence of IL-1beta or dexamethasone. The extent of collagen degradation was determined by measurement of the amount of hydroxyproline generated by acid-heat hydrolysis of culture supernatants. The expression of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) was evaluated by immunoblot analysis, gelatin zymography, and reverse transcription and real-time polymerase chain reaction. The phosphorylation of mitogen-activated protein kinases (MAPKs) in corneal fibroblasts was assessed by immunoblot analysis. RESULTS Dexamethasone inhibited IL-1beta-induced collagen degradation by corneal fibroblasts in a dose-dependent manner. Both the synthesis and activation of MMPs and the expression of TIMPs were inhibited by dexamethasone, as was the activity of plasmin in culture supernatants. Dexamethasone also inhibited the IL-1beta-induced phosphorylation of the MAPKs extracellular signal-regulated kinase (ERK) and c-Jun N-terminal kinase (JNK), but not that of p38. CONCLUSIONS Dexamethasone exerted multiple effects on the MMP-TIMP system in corneal fibroblasts and thereby inhibited IL-1beta-induced collagen degradation by these cells. Inhibition of the IL-1beta-induced activation of ERK and JNK may contribute to these effects of dexamethasone.