[1]蔡晖,石华宗,杨豫湘.过表达低氧诱导因子-1α（HIF-1α）对体外培养人视网膜血管内皮细胞增殖、炎症反应及血管生成的影响[J].眼科新进展,2019,39(1):032-35.[doi:10.13389/j.cnki.rao.2019.0007]
　CAI Hui,SHI Hua-Zong,YANG Yu-Xiang.The effects of over-expression of hypoxia inducible factor-1α on cell proliferation，inflammatory response and angiogenesis in vitro cultured human retinal capillary endothelial cells[J].Recent Advances in Ophthalmology,2019,39(1):032-35.[doi:10.13389/j.cnki.rao.2019.0007]
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过表达低氧诱导因子-1α（HIF-1α）对体外培养人视网膜血管内皮细胞增殖、炎症反应及血管生成的影响/HTML
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
39卷
期数:
2019年1期
页码:
032-35
栏目:
实验研究
出版日期:
2019-01-05

文章信息/Info

Title:
The effects of over-expression of hypoxia inducible factor-1α on cell proliferation，inflammatory response and angiogenesis in vitro cultured human retinal capillary endothelial cells
作者:
蔡晖; 石华宗; 杨豫湘
435000　湖北省黄石市，黄石市第二医院眼科（蔡晖，石华宗）；435000　湖北省黄石市，黄石市第五医院眼科（杨豫湘）
Author(s):
CAI Hui; SHI Hua-Zong; YANG Yu-Xiang
Department of Ophthalmology，the Second Hospital of Huangshi（CAI Hui，SHI Hua-Zong），Huangshi 435000，Hubei Province，China；Department of Ophthalmology，the Fifth Hospital of Huangshi（YANG Yu-Xiang），Huangshi 435000，Hubei Province，China
关键词:
低氧诱导因子-1α; 过表达; 人视网膜血管内皮细胞; 增殖; 炎症反应; 血管内皮生长因子
Keywords:
hypoxia inducible factor 1 alpha; over-expression; HREC; proliferation; inflammatory response; vascular endothelial growth factor
分类号:
R774
DOI:
10.13389/j.cnki.rao.2019.0007
文献标志码:
A
摘要:
目的　检测过表达低氧诱导因子-1α（hypoxia inducible factor 1 alpha，HIF-1α）对体外培养人视网膜血管内皮细胞（human retinal capillary endothelial cell，HREC）增殖、炎症反应及血管生成的影响。方法　将体外培养HREC转染人工合成的pEGFP-HIF-1α重组载体，并采用荧光定量PCR检测转染前后HREC中HIF-1α的表达变化。采用四甲基偶氮唑蓝比色法（MTT）、酶联免疫吸附实验（ELISA）和Western blot分别检测过表达HIF-1α对HREC增殖、炎症反应及血管生成的影响。结果　体外转染pEGFP-HIF-1α可显著增加HREC中HIF-1α mRNA 和蛋白表达。过表达HIF-1α可显著促进HREC增殖，HREC转染pEGFP-HIF-1α后24 h、48 h及72 h A值分别为：0.34±0.04、0.57±0.04、0.83±0.05，而HREC转染pEGFP-C1后24 h、48 h及72 h A值分别为：0.26±0.03、0.44±0.04、0.61±0.04，两者相比差异均有统计学意义（均为P＜0.05）。过表达HIF-1α可显著增加HREC中肿瘤坏死因子（tumor necrosis factor，TNF）-α、白细胞介素（interleukin，IL）-1β及IL-6的蛋白表达水平，TNF-α、IL-1β及IL-6蛋白表达分别为（2.63±0.57）ng·L-1、（1.94±0.41）ng·L-1、（5.32±0.83）ng·L-1，而转染pEGFP-C1后HREC中TNF-α、IL-1β及IL-6蛋白表达分别为（1.20±0.34）ng·L-1、（0.66±0.27）ng·L-1、（1.89±0.60）ng·L-1，两者相比差异均有统计学意义（均为P＜0.05）。过表达HIF-1α可显著增加HREC中血管内皮生长因子（vascular endothelial growth factor，VEGF）蛋白表达，转染pEGFP-HIF-1α后细胞VEGF蛋白的表达为6.72±0.96，而转染pEGFP-C1后细胞VEGF蛋白的表达为2.31±0.47，两者相比差异有统计学意义（P＜0.05）。结论　过表达HIF-1α可促进体外培养HREC增殖、炎症反应及血管的生成。
Abstract:
Objective　To detect the effects of over-expression of hypoxia inducible factor 1 alpha （HIF-1α） on cell proliferation，inflammatory response and angiogenesis in vitro cultured human retinal capillary endothelial cells （HREC）.Methods　In vitro cultured HREC was transfected with artificially synthesized pEGFP-HIF-1α recombinant vector，and quantitative RT-PCR was used to detect the expression change of HIF-1α in HREC.The effects of overexpression of HIF-1α on cell proliferation，inflammatory reaction and angiogenesis were detected by using 3-（4，5-dimethyl-2-thiazolyl）-2，5-diphenyl-2-H-tetrazolium bromide （MTT），enzyme linked immunosorbent assay （ELISA） and Western blots，respectively.Results　The mRNA and protein expression levels of HIF-1α were significant up-regulated in vitro cultured HREC after transfected with pEGFP-HIF-1α plasmid.Over-expression of HIF-1α markedly increased cell proliferation in vitro cultured HREC.The 24 h，48 h and 72 h A values in pEGFP-HIF-1α plasmid group was 0.34±0.04，0.57±0.04 and 0.83±0.05，while theirs A values in pEGFP-C1 vector group was 0.26±0.03，0.44±0.04 and 0.61±0.04 （all P＜0.05）.Over-expression of HIF-1α significantly increased the TNF-α，IL-1β and IL-6 levels in vitro cultured HREC.The levels of TNF-α，IL-1β and IL-6 in pEGFP-HIF-1α plasmid group was （2.63±0.57）ng·L-1，（1.94±0.41）ng·L-1 and （5.32±0.83）ng·L-1，while theirs expression in pEGFP-C1 vector group was （1.20±0.34）ng·L-1，（0.66±0.27）ng·L-1 and （1.89±0.60）ng·L-1，respectively （all P＜0.05）.Over-expression of HIF-1α notably increased the expression of vascular endothelial growth factor （VEGF） protein in vitro cultured HREC.The protein expression of VEGF in pEGFP-HIF-1α plasmid group was 6.72±0.96，while its expression in pEGFP-C1 vector group was 2.31±0.47，with significant difference （P＜0.05）.Conclusion　Over-expression of HIF-1α promotes cell proliferation，inflammatory response and angiogenesis in vitro cultured HREC.

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更新日期/Last Update: 2019-01-04