Zhang Le, Li Rong, Wu Bing-Hui, Liang Ting-Ting, Liu Zhe, Ju Wei, Wang Yi, Wen Yu-Ting, Liu Ming-Cui, Du Jun-Hui
Department of Ophthalmology, Northwest Woman's and Children's Hospital, Xi'an 710061, Shaanxi Province, China.
Department of Ophthalmology, Shaanxi Provincial People's Hospital, Xi'an 710068, Shaanxi Province, China.
Int J Ophthalmol. 2022 Apr 18;15(4):554-559. doi: 10.18240/ijo.2022.04.05. eCollection 2022.
To investigate the effect of leptin on the angiogenesis of RF/6A cells (monkey retinal choroidal endothelial cells) and test the cellular signaling in the mechanism.
RF/6A cells were cultured and randomly divided into four groups: normal control, with leptin at 50, 100, 200 ng/mL for cell counting kit-8 (CCK8). RF/6A cell proliferation and migration were examined by Transwell assays, while RF/6A cell tube formation by Matrigel assay. JAK2, p-JAK2, STAT3, and p-STAT3 protein expression was measured by Western blotting. Cells were then divided into the following treatment groups: control, 100 ng/mL leptin and AG-490 (100 ng/mL leptin+10 µmol/L AG-490) for examinations of RF/6A cellular behaviour again. Analysis of differences was carried out using one-way ANOVA and least significant difference (LSD).
RF/6A cell proliferation, migration and cell tube formation were promoted significantly by leptin in a dose-dependent manner (<0.05). Western blotting showed that leptin up-regulated p-JAK2 and p-STAT3 expression levels. Treatment with the JAK/STAT pathway inhibitor, AG-490, decreased leptin-induced p-JAK2 and p-STAT3 expression, and inhibited cell proliferation, migration and cell tube formation induced by leptin (<0.05).
Leptin can promote RF/6A cell angiogenesis activation of the JAK2/STAT3 signaling pathway.
研究瘦素对RF/6A细胞（猴视网膜脉络膜内皮细胞）血管生成的影响，并检测该机制中的细胞信号传导。
培养RF/6A细胞并随机分为四组：正常对照组，以及分别添加50、100、200 ng/mL瘦素的组，采用细胞计数试剂盒-8（CCK8）检测。通过Transwell实验检测RF/6A细胞增殖和迁移，采用基质胶实验检测RF/6A细胞管腔形成。通过蛋白质免疫印迹法检测JAK2、p-JAK2、STAT3和p-STAT3蛋白表达。然后将细胞分为以下处理组：对照组、100 ng/mL瘦素组和AG-490组（100 ng/mL瘦素+10 µmol/L AG-490），再次检测RF/6A细胞行为。采用单因素方差分析和最小显著差法（LSD）进行差异分析。
瘦素以剂量依赖性方式显著促进RF/6A细胞增殖、迁移和细胞管腔形成（<0.05）。蛋白质免疫印迹法显示，瘦素上调p-JAK2和p-STAT3表达水平。用JAK/STAT通路抑制剂AG-490处理可降低瘦素诱导的p-JAK2和p-STAT3表达，并抑制瘦素诱导的细胞增殖、迁移和细胞管腔形成（<0.05）。
瘦素可通过激活JAK2/STAT3信号通路促进RF/6A细胞血管生成。
