棕榈酰乙醇胺对全反式视黄醛诱导的人源视网膜色素上皮细胞退变的影响
Effect of palmitoylethanolamide on degeneration of a human-derived retinal pigment epithelial cell induced by all-trans retinal.
作者信息
Han Yun, Yang Kun-Huan, He Dan-Xue, Yu Chao-Feng, Tao Lei, Liao Chun-Yan, Cai Bin-Xiang, Liu Zu-Guo, Qiu Yan, Wu Ya-Lin
机构信息
Eye Institute of Xiamen University, Fujian Provincial Key Laboratory of Ophthalmology and Visual Science, Engineering and Research Center of Eye Regenerative Medicine, School of Medicine, Xiamen University, Xiamen 361102, Fujian Province, China.
Department of Ophthalmology, Xiang'an Hospital of Xiamen University, Xiamen 361102, Fujian Province, China.
出版信息
Int J Ophthalmol. 2023 Feb 18;16(2):191-200. doi: 10.18240/ijo.2023.02.04. eCollection 2023.
AIM
To study the effect of palmitoylethanolamide (PEA) on apoptosis of retinal pigment epithelial (RPE) cells induced by all-trans retinal (atRAL) and to explore the possible molecular mechanism.
METHODS
CellTiter 96 Aqueous One Solution Cell Proliferation Assay (MTS) was used to detect the effect of PEA on human-derived retinal epithelial cells (ARPE-19) viability induced by atRAL. A Leica DMi8 inverted microscope was used to observe cell morphology. Reactive oxygen species (ROS) production was evaluated with 2',7'-dichlorodihydrof-luorescein diacetate (H2DCFDA) staining and fluorescence microscopy. Expression of c-Jun N-terminal kinase (JNK), phosphorylated JNK (p-JNK), c-Jun, phosphorylated c-Jun (p-c-Jun), Bak, cleaved caspase-3, C/EBP homologous protein (CHOP), and binding (Bip) protein levels were tested by Western blot. mice, mouse models of atRAL clearance defects which displays some symbolic characteristics of dry age-related macular degeneration (AMD) and Stargardt disease (STGD1). In the animal models, PEA was injected intraperitoneally. The full-field electroretinogram was used to detect visual function under scotopic conditions traced from mice. Optical coherence tomography showed reconstitution or thickening of the retinal pigment epithelium layer. Effect of PEA on fundus injury induced by light in mice was observed by fundus photography.
RESULTS
PEA ameliorated ARPE-19 cells apoptosis and inhibited ROS (including mitochondrial ROS) production induced by atRAL. PEA improved the retinal functional, prohibited both RPE and photoreceptor from death, ameliorates light-induced fundus impairment in mice. and , PEA inhibited JNK, p-JNK, c-Jun, p-c-Jun, Bak, cleaved caspase-3, CHOP, and Bip protein levels induced by all-trans retinal in ARPE-19 cells.
CONCLUSION
PEA has effect on treating RPE cells apoptosis in retinopathy caused by atRAL accumulation. PEA is a potential treatment strategy for dry AMD and STGD1. The molecular mechanism is affecting the ROS-JNK-CHOP signaling pathway partly.
目的
研究棕榈酰乙醇酰胺（PEA）对全反式视黄醛（atRAL）诱导的视网膜色素上皮（RPE）细胞凋亡的影响，并探讨其可能的分子机制。
方法
采用CellTiter 96 Aqueous One Solution细胞增殖检测试剂盒（MTS）检测PEA对atRAL诱导的人源视网膜上皮细胞（ARPE-19）活力的影响。使用徕卡DMi8倒置显微镜观察细胞形态。用2',7'-二氯二氢荧光素二乙酸酯（H2DCFDA）染色和荧光显微镜评估活性氧（ROS）的产生。通过蛋白质免疫印迹法检测c-Jun氨基末端激酶（JNK）、磷酸化JNK（p-JNK）、c-Jun、磷酸化c-Jun（p-c-Jun）、Bak、裂解的半胱天冬酶-3、C/EBP同源蛋白（CHOP）和结合蛋白（Bip）的表达水平。选用呈现干性年龄相关性黄斑变性（AMD）和斯特格黄斑营养不良（STGD1）某些标志性特征的atRAL清除缺陷小鼠模型。在动物模型中，通过腹腔注射PEA。使用全视野视网膜电图检测暗视条件下小鼠的视觉功能。光学相干断层扫描显示视网膜色素上皮层的重建或增厚。通过眼底照相观察PEA对小鼠光诱导的眼底损伤的影响。
结果
PEA改善了ARPE-19细胞凋亡，并抑制了atRAL诱导的ROS（包括线粒体ROS）产生。PEA改善了视网膜功能，阻止了RPE和光感受器细胞死亡，减轻了小鼠光诱导的眼底损伤。此外，PEA抑制了atRAL诱导的ARPE-19细胞中JNK、p-JNK、c-Jun、p-c-Jun、Bak、裂解的半胱天冬酶-3、CHOP和Bip蛋白水平。
结论
PEA对治疗因atRAL积累引起的视网膜病变中的RPE细胞凋亡有作用。PEA是干性AMD和STGD1的一种潜在治疗策略。其分子机制部分是通过影响ROS-JNK-CHOP信号通路实现的。
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