山东大学耳鼻喉眼学报 ›› 2015, Vol. 29 ›› Issue (2): 74-80.doi: 10.6040/j.issn.1673-3770.0.2014.351

• 论著 • 上一篇    下一篇

尼古丁对人RPE细胞及HUVEC的影响

张营春, 杜祥阁, 颜昕, 王翠, 赵博军   

  1. 山东大学附属省立医院眼科中心, 山东 济南 250021
  • 收稿日期:2014-11-05 出版日期:2015-04-16 发布日期:2015-04-16
  • 作者简介:张营春。E-mail:jiningzyc@163.com
  • 基金资助:
    山东省自然科学基金资助项目(ZR2010HM098);山东省科技发展计划项目(2010G0020256)

Effect of nicotine on human RPE cells and HUVECs

ZHANG Ying-chun, DU Xiang-ge, YAN Xin, WANG Cui, ZHAO Bo-jun   

  1. Department of Ophthalmology, Shandong Provincial Hospital Affiliated to Shandong University, Jinan 250021, Shandong, China
  • Received:2014-11-05 Online:2015-04-16 Published:2015-04-16
  • Contact: 赵博军。E-mail:zhaobojun@hotmail.com E-mail:zhaobojun@hotmail.com

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
[1] Guymer R H, Chong E W. Modifiable risk factors for age-related macular degeneration[J]. Med J Aust, 2006, 184:455-458.
[2] 张营春, 赵博军. 烟草烟雾与老年黄斑变性[J].中华眼底病杂志, 2014,30(3):325-328. ZHANG Yingchun, ZHAO Bojun. Smoking and frog with aged age-related macular degeneration[J]. Chin J Ocular Fund Dis, 2014, 30(3):325-328.
[3] Zhao B J, Cai J, Boulton M. Expression of placenta growth factor is regulated by both VEGF and hyperglycaemia via VEGFR-2[J]. Microvasc Res, 2004, 68(3):239-246.
[4] Proulx S, Landreville S, Guerin S L, et al. Integrin alpha5 expression by the ARPE-19 cell line: comparison with primary RPE cultures and effect of growth medium on the alpha5 gene promoter strength[J]. Exp Eye Res, 2004, 79(2):157-165.
[5] Bull H A, Pittilo R M, Woolf N, et al. The effect of nicotine on human endothelial cell release of prostaglandins and ultrastructure[J]. Br J Exp Pathol, 1988, 69(3):413-421.
[6] Benowitz N L, Kuyt F, Jacob P. Influence of nicotine on cardiovascular and hormonal effects of cigarette smoking[J]. Clin Pharmacol, 1984, 36(1):74-81.
[7] Smith W, Mitchell P, Leeder S R. Smoking and age-related maculopathy. The Blue Mountains Eye Study[J]. Arch Ophthalmol, 1996, 114(12):1518-1523.
[8] Khan J C, Thurlby D A, Shahid H, et al. Smoking and age related macular degeneration: the number of pack years of cigarette smoking is a major determinant of risk for both geographic atrophy and choroidal neovascularisation[J]. Br J Ophthalmol, 2006, 90(1):75-80.
[9] Tomany S C, Wang J J, Van Leeuwen R, et al. Risk factors for incident age-related macular degeneration: pooled findings from 3 continents[J]. Ophthalmology, 2004, 111(7):1280-1287.
[10] Zhao B, A Ma, Cai J, et al. VEGF-A regulates the expression of VEGF-C in human retinal pigment epithelial cells[J]. Br J Ophthalmol, 2006, 90(8):1052-1059.
[11] Zhao B J, Smith G, Cai J, et al. Vascular endothelial growth factor C promotes survival of retinal vascular endothelial cells via vascular endothelial growth factor receptor-2[J]. Br J Ophthalmol, 2007, 91(4):538-545.
[12] Witmer A N, Vrensen G F, Van Noorden C J, et al. Vascular endothelial growth factors and angiogenesis in eye disease[J]. Prog Retin Eye Res, 2003, 22(1):1-29.
[13] Macklin K D, Maus A D, Pereira E F, et al. Human vascular endothelial cells express functional nicotinic acetylcholine receptors[J]. J Pharmacol Exp Ther, 1998, 287(1):435-439. [14] Park Y J, Lee T, Ha J, et al. Effect of nicotine on human umbilical vein endothelial cells (HUVECs) migration and angiogenesis[J]. Vascul Pharmacol, 2008, 49(1):32-36.
[15] Pans M, Marin-Castano M E. Nicotine increases the VEGF/PEDF ratio in retinal pigment epithelium: a possible mechanism for CNV in passive smokers with AMD[J]. IVOS, 2011, 52(6):3842-3853.
[16] Klettner A K, Doths J, Roider J. Nicotine reduces VEGF-secretion and phagocytotic activity in porcine RPE[J]. Graefes Arch Clin Exp Ophthalmol, 2012, 250(1):33-38.
[17] Gagat M, Grzanka D, Izdebska M, et al. Nornicotine impairs endothelial cell-cell adherens junction complexes in EA.hy926 cell line via structural reorganization of F-actin[J]. Folia Histochem Cytobiol, 2013, 51(3):179-192.
[18] Lopez P F, Sippy B D, Lambert H M, et al. Transdifferentiated retinal pigment epithelial cells are immunoreactive for vascular endothelial growth factor in surgically excised age-related macular degeneration-related choroidal neovascular membranes[J]. Invest Ophthalmol Vis Sci, 1996, 37(10):855-868.
[19] Kliffen M, Sharma H S, Mooy C M, et al. Increased expression of angiogenic growth factors in age-related maculopathy[J]. Br J Ophthalmol, 1997, 81(2):154-162.
[20] Bhutto I A, McLeod D S, Hasegawa T, et al. Pigment epithelium derived factor (PEDF) and vascular endothelial growth factor (VEGF) in aged human choroid and eyes with age-related macular degeneration[J]. Exp Eye Res, 2006, 82(1):99-110.
[21] Kanda Y, Watanabe Y. Nicotine-induced vascular endothelial growth factor release via the EGFR-ERK pathway in rat vascular smooth muscle cells[J]. Life Sci, 2007, 80(15):1409-1414.
[1] 李浩,李延忠,王岩. HIF-1α、VEGF在阻塞性睡眠呼吸暂停低通气综合征患者[J]. 山东大学耳鼻喉眼学报, 2018, 32(2): 43-47.
[2] 王翠,颜昕,赵博军. IVR联合PDT治疗湿性年龄相关性黄斑变性的临床观察[J]. 山东大学耳鼻喉眼学报, 2017, 31(4): 94-97.
[3] 周玮琰,王洪亚,杜秀娟,董卫红. 甘糖酯对糖尿病大鼠视网膜病变中血管生成因子VEGF表达的影响[J]. 山东大学耳鼻喉眼学报, 2017, 31(2): 90-95.
[4] 李盈盈, 周涵, 张伟强, 董伟达. 沉默HIF-1α基因对鼻黏膜上皮细胞生长因子表达的影响[J]. 山东大学耳鼻喉眼学报, 2015, 29(5): 38-42.
[5] 李俊英. 瑞舒伐他汀联合非诺贝特对老年糖尿病视网膜病变患者血管内皮功能的影响[J]. 山东大学耳鼻喉眼学报, 2015, 29(5): 72-75.
[6] 杜祥阁, 张营春, 颜昕, 王翠, 赵博军. 下调血管内皮细胞蛋白激酶CK2表达对血管增殖影响的体外研究[J]. 山东大学耳鼻喉眼学报, 2015, 29(3): 76-80.
[7] 龚斌, 范传峰, 王玉. 抗VEGF药物(雷珠单抗)治疗不同病程湿性年龄相关性黄斑变性62例[J]. 山东大学耳鼻喉眼学报, 2014, 28(5): 58-61.
[8] 訾雨梦1,王玉2,范传峰2,舒湘汶2,吴昌龙2. 玻璃体腔注射治疗不同病程两组特发性脉络膜新生血管病变[J]. 山东大学耳鼻喉眼学报, 2014, 28(3): 63-66.
[9] 谢丽综述,魏伟审校. 糖尿病性黄斑水肿的治疗进展[J]. 山东大学耳鼻喉眼学报, 2013, 27(2): 81-85.
[10] 李俐华,任基浩, 殷团芳,刘伟. 儿童复发性呼吸道乳头状瘤组织中STAT3、VEGF的表达及微血管密度测定与复发、侵袭的关系[J]. 山东大学耳鼻喉眼学报, 2011, 25(6): 11-.
[11] 李臻. 糖皮质激素对鼻息肉缺氧诱导因子-1α及血管内皮生长因子表达的影响[J]. 山东大学耳鼻喉眼学报, 2011, 25(1): 53-.
[12] 韩旭光1,2 ,吴欣怡1 ,曲飞3 ,孙眞2 ,王旭2 ,徐湘辉2
. 角膜缝线与碱烧伤诱导兔角膜
新生血管的实验观察
[J]. 山东大学耳鼻喉眼学报, 2009, 23(2): 79-82 .
[13] 高 英,潘新良,王 岩,刘平云 . 血管内皮生长因子在鼻息肉组织中的表达和意义[J]. 山东大学耳鼻喉眼学报, 2008, 22(2): 150-152 .
[14] 马晓华,毕宏生,李镜海,解孝锋,吴建峰,季 鹏 . 雌二醇对牛视网膜毛细血管内皮细胞VEGF、HIF-1α的调控[J]. 山东大学耳鼻喉眼学报, 2008, 22(1): 6-09 .
[15] 徐志华,王继群,张 涛,贾海英,孙 瑾,山艳春 . VEGF在喉癌中表达的Meta分析[J]. 山东大学耳鼻喉眼学报, 2008, 22(1): 32-34 .
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed   
No Suggested Reading articles found!