[1]李蓉,杜军辉,常远.肿瘤坏死因子-α对RF/6A细胞自噬促进细胞增殖、迁移和管腔形成的影响[J].眼科新进展,2015,35(12):1132-1136.[doi:10.13389/j.cnki.rao.2015.0310]
　LI Rong,DU Jun-Hui,CHANG Yuan.TNF-α promoting proliferation ,migration and tube formation of RF/6A cells by inducing cellular autophagy[J].Recent Advances in Ophthalmology,2015,35(12):1132-1136.[doi:10.13389/j.cnki.rao.2015.0310]
点击复制

肿瘤坏死因子-α对RF/6A细胞自噬促进细胞增殖、迁移和管腔形成的影响
分享到：

《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
35卷
期数:
2015年12期
页码:
1132-1136
栏目:
实验研究
出版日期:
2015-12-05

文章信息/Info

Title:
TNF-α promoting proliferation ,migration and tube formation of RF/6A cells by inducing cellular autophagy
作者:
李蓉; 杜军辉; 常远
710077　陕西省西安市，西安医学院第一附属医院眼科
Author(s):
LI Rong; DU Jun-Hui; CHANG Yuan
Department of Ophthalmology ,the First Affiliated Hospital of Xi ’ an Medical University , Xi ’an 710077 . Shaanxi Province . China
关键词:
血管生成; 视网膜新生血管; 炎症; 自噬; 肿瘤坏死因子α; Beclin-1
Keywords:
angiogenesis ;  retinal neovascularization;  inflammation ;  autophagy;  TNF-a ;  Beclin-1
DOI:
10.13389/j.cnki.rao.2015.0310
文献标志码:
A
摘要:
目的　本研究观察肿瘤坏死因子α（ｔｕｍｏｒｎｅｃｒｏｓｉｓｆａｃｔｏｒ-ａｌｐｈａ，ＴＮＦ-α）对体外培养的恒河猴脉络膜／视网膜内皮细胞（ＲＦ／６Ａ）表达自噬蛋白Ｂｅｃｌｉｎ-１及细胞增殖、迁移和管腔形成的影响，探讨ＴＮＦ-α参与新生血管生成的可能机制。方法　将生长良好的ＲＦ／６Ａ细胞随机分为空白对照组、ＴＮＦ-α组和ＴＮＦ-α＋３-ＭＡ组。培养２４ｈ、４８ｈ后采用Ｗｅｓｔｅｒｎｂｌｏｔ检测细胞Ｂｅｃ-ｌｉｎ-１蛋白的表达，ＭＴＴ法检测细胞增殖，细胞划痕法检测细胞迁移，Ｍａｔｒｉｇｅｌ法检测管腔形成。结果　培养２４ｈ和４８ｈ，各组Ｂｅｃｌｉｎ-１／β-ａｃｔｉｎ比值分别是ＴＮＦ-α组（２４ｈ）０．６７３±０．０１７、ＴＮＦ-α＋３-ＭＡ组（２４ｈ）０．４９１±０．０１７、ＴＮＦ-α组（４８ｈ）０．７９２±０．００６、ＴＮＦ-α＋３-ＭＡ组（４８ｈ）０．５０４±０．００７、空白对照组０．２６８±０．０１７。ＴＮＦ-α组ＲＦ／６Ａ细胞Ｂｅｃｌｉｎ-１的表达量均明显高于空白对照组（Ｐ＜０．０５），ＴＮＦ-α＋３-ＭＡ组Ｂｅｃｌｉｎ-１的表达量较ＴＮＦ-α组明显减少（Ｐ＜０．０５）。各组细胞的相对增殖率分别是ＴＮＦ-α组（２４ｈ）１．４１０±０．０１０、ＴＮＦ-α＋３-ＭＡ组（２４ｈ）１．２９０±０．００４、ＴＮＦ-α组（４８ｈ）１．３２０±０．０１１、ＴＮＦ-α＋３-ＭＡ组（４８ｈ）０．１８０±０．０１５、对照组（２４ｈ）１．０００±０．０２０、对照组（４８ｈ）１．０００±０．０１１；ＴＮＦ-α组细胞的相对增殖率均较空白对照组升高（Ｐ＜０．０５），３-ＭＡ预处理后，ＴＮＦ-α促进ＲＦ／６Ａ细胞增殖的能力在两个时间点均受到抑制（均为Ｐ＜０．０５）。各组细胞的相对迁移距离分别是ＴＮＦ-α组（２４ｈ）（３４５±２８）μｍ、ＴＮＦ-α＋３-ＭＡ组（２４ｈ）（２５９±７７）μｍ、ＴＮＦ-α组（４８ｈ）（７６２±５５）μｍ、ＴＮＦ-α＋３-ＭＡ组（４８ｈ）（６５９±４８）μｍ、空白对照组（２４ｈ）（１９５±６３）μｍ、空白对照组（４８ｈ）（４１２±９４）μｍ；ＴＮＦ-α处理组ＲＦ／６Ａ细胞２４ｈ的迁移明显强于对照组（Ｐ＜０．０５），加入３-ＭＡ预处理后，这种增强作用有所下降，但仍然明显高于对照组（Ｐ＜０．０５）；培养４８ｈ，更多细胞迁移入划痕区域，较２４ｈ明显增加；不同组之间的差异与２４ｈ时的结果类似，差异有统计学意义（Ｐ＜０．０５）；培养２４ｈ各组细胞管腔形成个数：ＴＮＦ-α组（１１．８０±０．８１）个、ＴＮＦ-α＋３-ＭＡ组（７．５０±０．７２）个、空白对照组（４．３０±１．１２）个，ＴＮＦ-α组及ＴＮＦ-α＋３-ＭＡ组管腔形成个数均高于对照组（均为Ｐ＜０．０５），ＴＮＦ-α＋３-ＭＡ组较ＴＮＦ-α组明显减少（Ｐ＜０．０５）。结论　ＴＮＦ-α能够促进ＲＦ／６Ａ细胞的增殖、迁移和管腔样结构的形成，且ＴＮＦ-α促进内皮细胞自噬在血管新生中发挥重要的调控作用。
Abstract:
Objective To observe the effects of tumor necrosis factor-alpha ( TNF-α ) on the expression of autophagic protein Beclin-I and cell proliferation , migration and tube formation of a thesus macaque choroid-retinal endothelial ( RF/6A) cells . and explore the preliminary mechanism of the role of TNF-α in retinal angiogenesis. Methods Cultured RF/6A cells were randomly divided into the control group ,TNF-a group and TNF-α + 3-MA group according to the different treatment in culture medium. After 24 hours and 48 hours.Beclin-l was tested by Western blot analysis and cell proliferation,migration and tube formation was detected by MTT assay, scratch assay and matrigel, respectively. Results After 24 hours and 48 hours , the ratios of Beclin-I/[3actin were 0. 673 + 0. 017 ( TNF-a group , 24 hours ) , 0. 491 + 0. 017 ( TNF-α + 3-MA group , 24 hours ) .0. 792 + 0. 006 ( TNF-α group ,48 hours) .0. 504 +0. 007 ( TNF-α + 3-MA group ,48 hours) and 0. 268 +0. 017 ( control ~oup) , respectively. Expression of Beclin-1 by RF/6A cells in TNF-a group for both time points was sigruficantly higher than the control group (P < 0. 05 ) and TNF-ce + 3-MA group (P < 0. 05 ) . The cell relative growth rate ( RGR) were l. 410 + 0. 010 ( TNF-α group , 24 hours ) . 1. 290 + 0. 004 ( TNF-α + 3-MA group , 24 hours) , 1. 320 +0. 011 ( TNF-α group ,48 hours ) .0. 180 + 0. 015 ( TNF-α + 3-MA group ,48 hours) , 1. 000 + 0. 020 ( control group ,24 hours ) and l. 000 + 0. 011 ( control group,48 hours) , respectively. RGR in TNF-α group was higher than the control group (P < 0. 05 ) . And the promoting effects of TNF-α on RGR of RF/6A cells was inhibited for both time points when pretreated with 3-MA (P < 0. 05 ) . The migration distance were ( 345 + 28 ) ym ( TNF-α group, 24 hours) , ( 259 + 77 ) ym ( TNF-α + 3-MA group ,24 hours ) , ( 762 + 55 ) ym ( TNF-α group ,48 hours ) , ( 659 + 48 ) ym ( TNF-α + 3MA group ,48 hours) , ( 195 + 63 ) ym ( control group , 24 hours ) and (412 + 94 ) Vm ( control group,48 hours) , respectively. The migration distance of RF/6A cells in the TNF-a group was siguficantly longer than the control group (P < 0. 05 ) , and it was decreased when pretreated with 3-MA,but still longer than the control group (P < 0. 05 ) at 24 hours time point. Compared with 24 hours. more cells migrated into the scratch areas at 48 hours and the differences among the three groups were similar to that of 24 hours (P < 0. 05 ) . The number of tube formation at 24h werell. 80 +0. 81 ( TNF-α group) ,7. 50 +0. 72 ( TNF-α + 3-MA group ) and 4. 30 + 1. 12 ( control group ) , respectively. The number in TNF-α and TNF-α +3-MA groups was higher than the control group ( P < 0. 05 ) . and that in TNF-α + 3-MA group was lower than TNF-α group ( P < 0. 05 ) . Conclusion TNF-α can promote proliferation ,migration and tube formation of RF/6A cells and induction of autophagy , and TNF-α plays an important role in regulating angiogenesis.

相似文献/References:

[1]迟源 王海林 谢晚晴 才文卓 才丽平.MicroRNA调节视网膜新生血管研究进展[J].眼科新进展,2013,33(2):000.
[2]耿金 陈蕾 柳力敏 刘宁宁 刘磊.β-榄香烯抑制氧诱导小鼠视网膜新生血管形成的实验研究[J].眼科新进展,2013,33(3):000.
[3]杨玉新 李士清 周焕娇 李涛.一种精确评估氧诱导小鼠视网膜新生血管和无灌注区的联合方法[J].眼科新进展,2013,33(6):000.
[4]王淋淋 唐兰芬 谭建新 阮豫才 方玉兰.曲尼司特抑制早产儿视网膜病变大鼠模型血管新生和纤维化的实验研究[J].眼科新进展,2013,33(10):000.
[5]贺玲 郑晓龙 张杰 张善峰 余晓红 宋海云.熊果酸对氧诱导小鼠视网膜病变的保护作用[J].眼科新进展,2013,33(10):000.
[6]韩丽英 李兵.IGF-1对大鼠未成熟视网膜新生血管形成的影响[J].眼科新进展,2013,33(10):000.
[7]史少阳 张媛 李迅 裴存文 陈晓隆.Dll4、Notch1在大鼠氧诱导性视网膜病变模型中的表达[J].眼科新进展,2013,33(10):000.
[8]王龙梅 闫琳 杨侠 董晓光 徐海峰. 高氧对小鼠视网膜新生血管模型中根蛋白的影响[J].眼科新进展,2014,34(3):217.
[9]高翔,王雨生. 免疫相关的巨噬细胞和髓样血管生成细胞在视网膜新生血管发生发展中的作用[J].眼科新进展,2014,34(7):687.[doi:10.13389/j.cnki.rao.2014.0189]
[10]刘昳,袁进.核转录因子-κB在氧诱导血管增殖性视网膜病变小鼠中的表达[J].眼科新进展,2014,34(12):1101.[doi:10.13389/j.cnki.rao.2014.0306]
　LIU Yi,YUAN Jin.Expression of nuclear factor-kappa B in mice with oxygen-induced proliferative retinopathy[J].Recent Advances in Ophthalmology,2014,34(12):1101.[doi:10.13389/j.cnki.rao.2014.0306]
[11]谢坤鹏,王新,杜军辉,等.脂联素对RF/6A细胞增殖、迁移及管腔形成的影响[J].眼科新进展,2017,37(6):511.[doi:10.13389/j.cnki.rao.2017.0129]
　XIE Kun-Peng,WANG Xin,DU Jun-Hui,et al.Effects of adiponectin on proliferation，migration and tube formation of RF/6A cells[J].Recent Advances in Ophthalmology,2017,37(12):511.[doi:10.13389/j.cnki.rao.2017.0129]
[12]李蓉,姚杨,杜军辉,等.自噬在高糖条件下视网膜色素上皮细胞表达血管内皮生长因子促进RF/6A细胞血管生成中的作用[J].眼科新进展,2019,39(8):714.[doi:10.13389/j.cnki.rao.2019.0163]
　LI Rong,YAO Yang,DU Jun-Hui,et al.Autophagy promotes angiogenesis of RF/6A cells following upregulating the expression of vascular endothelial growth factor in retinal pigment epithelial cells under high glucose conditions[J].Recent Advances in Ophthalmology,2019,39(12):714.[doi:10.13389/j.cnki.rao.2019.0163]

备注/Memo

备注/Memo:
陕西省教育厅科研基金资助（编号：１５ＪＫ１６２４）；陕西省科技厅科研基金资助（编号：２０１３ＪＣ２-１９）
更新日期/Last Update: 2015-12-07