Li Sijia, Su Dongmei, Hu Shanshan, Hu Qiang, Sun Dawei
Harbin Medical University, Harbin 150086, China; Hongqi Hospital of Mudanjiang Medical University, Mudanjiang 157011, Heilongjiang, China.
Department of Genetics, NHC Key Laboratory of Reproductive Health Engineering Technology Research, National Research Institute for Family Planning, Health Department, Beijing 100081, China; Graduate School, Peking Union Medical College, Beijing 100081, China.
Toxicol Appl Pharmacol. 2025 Feb;495:117124. doi: 10.1016/j.taap.2024.117124. Epub 2024 Dec 10.
Age-related macular degeneration (AMD) is a representative age-related ophthalmic disease, and the pathogenesis of AMD remains unclear. This research intended to determine whether epigallocatechin gallate (EGCG) could alleviate the progression of AMD and the possible mechanism. We constructed three groups of mice (young, aged, and EGCG), and HE and TUNEL staining of retinal tissues was performed to observe the structural changes in the retinal pigment epithelial (RPE) layer and the level of apoptosis, respectively. Through RNA-Sequencing analysis of retinal tissues and by RT-qPCR, GO, KEGG, and literature analyses, we identified cytoplasmic fragile X mental retardation 1-interacting protein 2 (CYFIP2) as a possible effector gene for EGCG action and validated its role by immunofluorescent and western blotting experiments. The CCK-8 and Hoechst 33342 apoptosis assays, and western blotting and qRT-PCR assays showed that EGCG reduced hydrogen peroxide (HO)-induced apoptosis in adult human RPE (ARPE-19) cells, and the expression of Cyfip2 was changed accordingly. RNA interference analysis indicated that Cyfip2 knockdown alleviated HO-induced ARPE apoptosis, while its overexpression weakened EGCG's protective effect. Western blot analysis showed that Cyfip2 mediated the anti-apoptotic effect of EGCG by modulating the level of protein kinase B (Akt) phosphorylation in ARPE cells, and the activation level of phosphorylated AKT (p-AKT Ser473) in retinal tissue of the EGCG-fed group was higher than that of the aged group. Taken together, this study suggests that EGCG plays a protective role in the development of AMD and the apoptosis of ARPE cells through the Cyfip2/AKT pathway.
年龄相关性黄斑变性（AMD）是一种典型的年龄相关性眼科疾病，其发病机制尚不清楚。本研究旨在确定表没食子儿茶素没食子酸酯（EGCG）是否能缓解AMD的进展及其可能的机制。我们构建了三组小鼠（年轻组、老年组和EGCG组），并对视网膜组织进行苏木精-伊红（HE）染色和末端脱氧核苷酸转移酶介导的缺口末端标记（TUNEL）染色，分别观察视网膜色素上皮（RPE）层的结构变化和细胞凋亡水平。通过对视网膜组织进行RNA测序分析，并结合逆转录定量聚合酶链反应（RT-qPCR）、基因本体论（GO）、京都基因与基因组百科全书（KEGG）及文献分析，我们确定细胞质脆性X智力低下蛋白1相互作用蛋白2（CYFIP2）为EGCG作用的一个可能效应基因，并通过免疫荧光和蛋白质印迹实验验证了其作用。细胞计数试剂盒-8（CCK-8）和Hoechst 33342凋亡检测以及蛋白质印迹和qRT-PCR检测表明，EGCG可减少过氧化氢（HO）诱导的成人视网膜色素上皮（ARPE-19）细胞凋亡，且CYFIP2的表达也相应改变。RNA干扰分析表明，敲低CYFIP2可减轻HO诱导的ARPE细胞凋亡，而其过表达则削弱了EGCG的保护作用。蛋白质印迹分析表明，CYFIP2通过调节ARPE细胞中蛋白激酶B（Akt）的磷酸化水平介导了EGCG的抗凋亡作用，且EGCG喂养组视网膜组织中磷酸化AKT（p-AKT Ser473）的激活水平高于老年组。综上所述，本研究表明EGCG通过CYFIP2/Akt途径在AMD的发展和ARPE细胞凋亡中发挥保护作用。
