尼古丁对人RPE细胞及HUVEC的影响

doi:10.6040/j.issn.1673-3770.0.2014.351

摘要/Abstract

摘要： 目的探讨尼古丁对人视网膜色素上皮(RPE)细胞和血管内皮细胞(HUVECs)中血管内皮生长因子(VEGF)表达及形成新生血管能力的影响。方法 MTT法检测尼古丁对两种细胞增殖的影响;划痕及成管实验检测其对HUVECs迁移、成管的影响;RT-PCR和Western blotting法检测两种细胞经尼古丁处理后VEGF、PEDF mRNA及蛋白的表达。结果尼古丁促进HUVECs增殖、迁移、成管(P<0.05), 但对ARPE-19细胞增殖无影响(P>0.05), 且高浓度(100 μmol/L)时抑制细胞增殖(P<0.01)。尼古丁以剂量和时间依赖方式促进两种细胞VEGF mRNA及蛋白的表达, 抑制PEDF mRNA及蛋白的表达, 上调VEGF/PEDF基因表达的比率(P<0.05)。结论尼古丁可上调ARPE-19细胞和HUVECs中VEGF/PEDF基因表达的比率, 促进HUVECs增殖、迁移、成管。

关键词: 尼古丁, 人视网膜色素上皮细胞, 色素上皮衍生因子, 血管内皮细胞, 脉络膜新生血管, 血管内皮生长因子

Abstract: Objective To evaluate the influences of Nicotine (NT) from cigarette smoke on VEGF and PEDF expressions in retinal pigment epithelium (RPE) cells and human umbilical vein endothelial cells (HUVECs). In addition, the angiogenic behaviors of endothelial cells with NT treatment were assessed by using in vitro methods. Methods ARPE-19 cells and HUVECs were treated with different concentrations of NT for a period of times. Proliferative effect was investigated by using the method of MTT analysis. HUVECs migration and tube formation were assessed by using the wound-healing and Matrigel models. The expressions of VEGF and PEDF in both types of cells were examined by real-time PCR and Western blotting. Results There was no proliferation of ARPE-19 cells following treatment with various concentrations of NT. However, NT significantly stimulated HUVECs proliferation, migration, and tube formation. NT up-regulated the expression of VEGF but suppressed the expression of PEDF at both mRNA and protein levels in a dose- and time-dependent manner in ARPE-19 cells and HUVECs. Conclusion Our results demonstrate that NT promoted endothelial cellular proliferation, migration and angiogenesis of HUVECs in vitro. These effects might partly play through simultaneous modulation of VEGF/PEDF signaling in ARPE-19 cells and HUVECs.

Key words: Nicotine, Vascular endothelial growth factor, Pigment epithelium-derived factor, Vascular endothelial cells, Choroidal neovascularization, Human retinal pigment epithelium cells

中图分类号:

R774.1

张营春, 杜祥阁, 颜昕, 王翠, 赵博军. 尼古丁对人RPE细胞及HUVEC的影响[J]. 山东大学耳鼻喉眼学报, 2015, 29(2): 74-80.

ZHANG Ying-chun, DU Xiang-ge, YAN Xin, WANG Cui, ZHAO Bo-jun. Effect of nicotine on human RPE cells and HUVECs[J]. JOURNAL OF SHANDONG UNIVERSITY (OTOLARYNGOLOGY AND OPHTHALMOLOGY), 2015, 29(2): 74-80.

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