Zhang Hui-Yan, Wang Jian-Yong, Yao Hang-Ping
Hangzhou Vocational & Technical College, Hangzhou 310018, Zhejiang Province, China.
Department of Ophthalmology, the First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310003, Zhejiang Province, China.
Int J Ophthalmol. 2014 Jun 18;7(3):408-12. doi: 10.3980/j.issn.2222-3959.2014.03.04. eCollection 2014.
To investigate the mechanism underlying the anti-inflammatory effects of epigallocatechin-3-gallate (EGCG) in lipopolysaccharide (LPS)-stimulated human retinal endothelial cells (HRECs).
HRECs pre-treated with EGCG (0-100 µmol/L) were stimulated with LPS (250 ng/mL). Levels of tumor necrosis factor alpha (TNF-α), vascular endothelial growth factor (VEGF), monocyte chemotactic protein-1 (MCP-1) and nitric oxide (NO) in the supernatants were determined by enzyme-linked immunosorbent assay (ELISA) and Griess assay. The protein expression of phosphorylated extracellular signal-regulated kinase (ERK) 1/2 and p38 mitogen-activated protein kinases (p38) were determined by Western blot analysis.
EGCG pre-treatment significantly inhibited the secretion of TNF-α, VEGF, MCP-1 and NO in LPS-stimulated HRECs. Moreover, EGCG effectively attenuated LPS-induced activation and phosphorylation of ERK1/2 and p38 in HRECs in a dose-dependent manner.
EGCG exhibited inhibitory effects on LPS-induced pro-inflammatory cytokines production by modulating ERK1/2 and p38 pathways in HRECs, suggesting EGCG as a potential candidate for anti-inflammatory intervention.
研究表没食子儿茶素-3-没食子酸酯（EGCG）在脂多糖（LPS）刺激的人视网膜内皮细胞（HRECs）中发挥抗炎作用的机制。
用EGCG（0 - 100 μmol/L）预处理HRECs，然后用LPS（250 ng/mL）刺激。通过酶联免疫吸附测定（ELISA）和格里斯测定法测定上清液中肿瘤坏死因子α（TNF-α）、血管内皮生长因子（VEGF）、单核细胞趋化蛋白-1（MCP-1）和一氧化氮（NO）的水平。通过蛋白质印迹分析测定磷酸化细胞外信号调节激酶（ERK）1/2和p38丝裂原活化蛋白激酶（p38）的蛋白表达。
EGCG预处理显著抑制LPS刺激的HRECs中TNF-α、VEGF、MCP-1和NO的分泌。此外，EGCG以剂量依赖的方式有效减弱LPS诱导的HRECs中ERK1/2和p38的激活和磷酸化。
EGCG通过调节HRECs中的ERK1/2和p38途径对LPS诱导的促炎细胞因子产生具有抑制作用，表明EGCG作为抗炎干预的潜在候选物。
