Anti-inflammatory effect of docosahexaenoic acid on cytokine-induced adhesion molecule expression in human retinal vascular endothelial cells
Anti-inflammatory effect of docosahexaenoic acid on cytokine-induced adhesion molecule expression in human retinal vascular endothelial cells
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DOI：
10.1167/iovs.05-0601
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发表时间：
2005-11-01
影响因子：
4.4
通讯作者：
Busik, JV
中科院分区：
文献类型：
--
作者：
Chen, WQ;Esselman, WJ;Busik, JV
目的。二十二碳六烯酸(DHA)(22：6n3)是视网膜中主要的n3-多不饱和脂肪酸(PUFA)，许多体内和体外研究表明，DHA具有显著的抗炎作用。尽管血管炎症在糖尿病视网膜病变中具有重要作用，但DHA(22：6n3)对细胞因子刺激的人视网膜血管内皮细胞(HRVECs)的抗炎作用尚未被阐明。方法：用Western印迹法检测细胞因子诱导的细胞黏附分子(CAM)的表达。用Western印迹和凝胶迁移率改变分析(EMSA)分析DHA(22：6n3)对细胞因子诱导的核因子-kappaB信号转导的影响。结果血管内皮细胞生长因子(165)、肿瘤坏死因子α和IL-1β刺激hRVEC 6~24小时后，细胞内黏附分子(ICAM)-1和血管细胞黏附分子(VCAM)-1表达明显增加。用100mU的BSA结合的DHA(22：6n3)处理细胞24小时，可显著抑制细胞因子诱导的CAM表达。IL-1β、肿瘤坏死因子α和血管内皮生长因子(165)可诱导p65和p50核转录因子-kappaB异构体与VCAM-1启动子结合。DHA(22：6n3)可抑制细胞因子诱导的核因子-kappaB结合25%~40%。此外，DHA(22：6n3)可抑制IL-1β诱导的核因子抑制因子-kappa Bα(I-kappa Bα)的磷酸化，从而阻止其降解。结论IL-1β、肿瘤坏死因子α和血管内皮生长因子(165)通过激活核因子-kappa B途径诱导hRVECs表达CAM。DHA(22：6n3)通过抑制核因子-kappa B核转位和上游I-kappaBα的磷酸化和降解，抑制细胞因子诱导的细胞间黏附分子表达。DHA(22：6n3)在糖尿病视网膜细胞因子产生增加和CAM表达增加的情况下，可能是一种重要的抗炎药。
PURPOSE. Docosahexaenoic acid (DHA(22: 6n3)), the principal n3-polyunsaturated fatty acid (PUFA) in the retina, has been shown to have a pronounced anti-inflammatory effect in numerous in vivo and in vitro studies. Despite the importance of vascular inflammation in diabetic retinopathy, the anti-inflammatory role of DHA(22: 6n3) in cytokine-stimulated human retinal vascular endothelial cells (hRVECs) has not been addressed.METHODS. Cytokine-induced expression of cell adhesion molecules (CAMs) was assessed by Western blot. The effect of DHA(22: 6n3) on cytokine-induced nuclear factor (NF)-kappa B signaling was analyzed by Western blot analysis and electrophoretic mobility shift assay ( EMSA).RESULTS. Stimulation of hRVECs with VEGF(165), TNF alpha, or IL-1 beta for 6 to 24 hours caused significant induction of intracellular adhesion molecule ( ICAM)-1 and vascular cell adhesion molecule ( VCAM)-1 expression. Pretreatment of the cells with 100 mu M of BSA-bound DHA(22: 6n3) for 24 hours remarkably inhibited cytokine-induced CAM expression. IL-1 beta, TNF alpha, and VEGF(165) induced nuclear translocation and binding of p65 and p50 NF-kappa B isoforms to the VCAM-1 promoter. DHA(22: 6n3) pretreatment inhibited cytokine-induced NF-kappa B binding by 25% to 40%. Moreover, DHA(22: 6n3) diminished IL-1 beta induced phosphorylation of the inhibitor of nuclear factor (NF)-kappa B (I-kappa B alpha), thus preventing its degradation.CONCLUSIONS. IL-1 beta, TNF alpha, and VEGF(165) induced CAM expression in hRVECs through activation of the NF-kappa B pathway. DHA(22: 6n3) inhibited cytokine induced CAM expression through suppression of NF-kappa B nuclear translocation and upstream I-kappa B alpha phosphorylation and degradation. DHA(22: 6n3) could be an important anti-inflammatory agent in the face of increased cytokine production and CAM expression in the diabetic retina.