Mukherjee Pranab K, Marcheselli Victor L, Serhan Charles N, Bazan Nicolas G
Neuroscience Center of Excellence and Department of Ophthalmology, Louisiana State University Health Sciences Center School of Medicine, New Orleans, LA 70112, USA.
Proc Natl Acad Sci U S A. 2004 Jun 1;101(22):8491-6. doi: 10.1073/pnas.0402531101. Epub 2004 May 19.
Docosahexaenoic acid (DHA) is a lipid peroxidation target in oxidative injury to retinal pigment epithelium (RPE) and retina. Photoreceptor and synaptic membranes share the highest content of DHA of all cell membranes. This fatty acid is required for RPE functional integrity; however, it is not known whether specific mediators generated from DHA contribute to its biological significance. We used human ARPE-19 cells and demonstrated the synthesis of 10,17S-docosatriene [neuroprotectin D1 (NPD1)]. This synthesis was enhanced by the calcium ionophore A-23187, by IL-1beta, or by supplying DHA. Under these conditions, there is a time-dependent release of endogenous free DHA followed by NPD1 formation, suggesting that phospholipase A(2) releases the mediator's precursor. Added NPD1 potently counteracted H(2)O(2)/tumor necrosis factor alpha oxidative-stress-triggered apoptotic RPE DNA damage. NPD1 also up-regulated the antiapoptotic proteins Bcl-2 and Bcl-x(L) and decreased proapoptotic Bax and Bad expression. Moreover, NPD1 (50 nM) inhibited oxidative-stress-induced caspase-3 activation. NPD1 also inhibited IL-1beta-stimulated expression of cyclooxygenase 2 promoter transfected into ARPE-19 cells. Overall, NPD1 protected RPE cells from oxidative-stress-induced apoptosis, and we predict that it will similarly protect neurons. This lipid mediator therefore may indirectly contribute to photoreceptor cell survival as well. Because both RPE and photoreceptor cells die in retinal degenerations, our findings contribute to the understanding of retinal cell survival signaling and potentially to the development of new therapeutic strategies.
二十二碳六烯酸(DHA)是视网膜色素上皮(RPE)和视网膜氧化损伤中的脂质过氧化靶点。光感受器和突触膜在所有细胞膜中DHA含量最高。这种脂肪酸是RPE功能完整性所必需的;然而,尚不清楚DHA产生的特定介质是否有助于其生物学意义。我们使用人ARPE-19细胞并证明了10,17S-二十二碳三烯[神经保护素D1(NPD1)]的合成。钙离子载体A-23187、IL-1β或提供DHA可增强这种合成。在这些条件下,内源性游离DHA会随时间释放,随后形成NPD1,这表明磷脂酶A(2)释放了介质的前体。添加的NPD1能有效对抗H(2)O(2)/肿瘤坏死因子α氧化应激引发的RPE细胞凋亡性DNA损伤。NPD1还上调了抗凋亡蛋白Bcl-2和Bcl-x(L),并降低了促凋亡蛋白Bax和Bad的表达。此外,NPD1(50 nM)抑制氧化应激诱导的caspase-3激活。NPD1还抑制转染到ARPE-19细胞中的环氧合酶2启动子的IL-1β刺激表达。总体而言,NPD1保护RPE细胞免受氧化应激诱导的凋亡,我们预测它也会同样保护神经元。因此,这种脂质介质可能也间接有助于光感受器细胞的存活。由于RPE和光感受器细胞在视网膜变性中都会死亡,我们的发现有助于理解视网膜细胞存活信号,并可能有助于开发新的治疗策略。