Inhibitory effect of triptolide on chemokine expression induced by proinflammatory cytokines in human corneal fibroblasts
Inhibitory effect of triptolide on chemokine expression induced by proinflammatory cytokines in human corneal fibroblasts
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DOI：
10.1167/iovs.05-0010
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发表时间：
2005-07-01
影响因子：
4.4
通讯作者：
Nishida, T
中科院分区：
文献类型：
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作者：
Lu, Y;Fukuda, K;Nishida, T
目的。各种趋化因子的合成，包括白细胞介素（IL）-8和单核细胞趋化蛋白（MCP）-1，以及角膜成纤维细胞中细胞间粘附分子（ICAM）-1的表面表达，导致角膜炎症。与地塞米松相比，检查了雷公藤甲素（雷公藤草提取物的主要成分）对人角膜成纤维细胞中这些蛋白质表达的影响。方法。用酶联免疫吸附测定法测量培养的角膜成纤维细胞中IL-8和MCP-1的释放以及ICAM-1的表面表达。通过逆转录和实时聚合酶链式反应分析确定这些蛋白质 mRNA 的细胞丰度。通过用分泌型碱性磷酸酶报告基因进行细胞转染来评估转录因子 NF-κ B 和 AP-1 的活性。结果。雷公藤内酯醇和地塞米松均以剂量依赖性方式抑制促炎细胞因子IL-1β或肿瘤坏死因子(TNF)-α诱导的角膜成纤维细胞中IL-8和MCP-1的表达。这些抑制作用在 mRNA 和蛋白质水平上都很明显。两种化合物对细胞因子诱导的 ICAM-1 表达也具有较小的抑制作用。 IL-1β对NF-κB的激活被雷公藤内酯醇和地塞米松均显着抑制，而AP-1的活性不受这两种药物的影响。结论。与地塞米松一样，雷公藤甲素可抑制暴露于促炎细胞因子的培养人角膜成纤维细胞中的 IL-8 和 MCP-1 表达，这一作用很可能是通过抑制 NF-κ B 激活介导的。雷公藤内酯醇在体内的类似作用有望限制中性粒细胞和单核细胞向角膜的浸润。
Purpose. Synthesis of various chemokines, including interleukin (IL)-8 and monocyte chemotactic protein (MCP)-1, as well as the surface expression of intercellular adhesion molecule (ICAM)-1 in corneal fibroblasts contribute to corneal inflammation. The effects of triptolide, the major constituent of extracts of the herb Tripterygium wilfordii hook f, on the expression of these proteins in human corneal fibroblasts were examined in comparison with those of dexamethasone.Methods. The release of IL-8 and MCP-1 from and the surface expression of ICAM-1 on cultured corneal fibroblasts were measured with enzyme-linked immunosorbent assays. The cellular abundance of the mRNAs for these proteins was determined by reverse transcription and real-time polymerase chain reaction analysis. The activities of the transcription factors NF-kappa B and AP-1 were assessed by cell transfection with secretory alkaline phosphatase reporter genes.Results. Both triptolide and dexamethasone inhibited in a dose-dependent manner the expression of IL-8 and MCP-1 in corneal fibroblasts induced by the proinflammatory cytokines IL-1 beta or tumor necrosis factor (TNF)-alpha. These inhibitory effects were apparent at both the mRNA and protein levels. Both compounds also had a lesser inhibitory effect on cytokine-induced ICAM-1 expression. The activation of NF-kappa B by IL-1 beta was markedly inhibited by both triptolide and dexamethasone, whereas the activity of AP-1 was not affected by either agent.Conclusions. Like dexamethasone, triptolide inhibited IL-8 and MCP-1 expression in cultured human corneal fibroblasts exposed to proinflammatory cytokines, an action most likely mediated by inhibition of NF-kappa B activation. Similar effects of triptolide in vivo may be expected to limit the infiltration of neutrophils and monocytes into the cornea.