| 罗雪兰,陈 伟,莫国君,杨 鹏,欧和生.27-nt miRNA对血管内皮细胞eNOS表达/活性的调节及其代谢产物的影响[J].现代生物医学进展英文版,2017,17(8):1442-1447. |
| 27-nt miRNA对血管内皮细胞eNOS表达/活性的调节及其代谢产物的影响 |
| The Regulation of 27-nt miRNA on Vascular Endothelial Cells eNOS Expression/Activity and the Impact of Its Metabolites |
| Received:May 20, 2016 Revised:June 18, 2016 |
| DOI：10.13241/j.cnki.pmb.2017.08.009 |
| 中文关键词: 27-nt miRNA 内皮细胞 eNOS 代谢产物 |
| 英文关键词: 27-nt miRNA Endothelial Cells ENOS Metabolite |
| 基金项目:国家自然科学基金面上项目（81373403）；广西硕士研究生科研创新项目（自然科学）（YCSZ2015119） |
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| 中文摘要: |
| 摘要 目的：前期工作表明，27-nt miRNA对eNOS 的转录和表达有负反馈调节作用。本实验进一步探讨27-nt miRNA 对血管内皮细胞eNOS的基因表达、活性调节及其代谢产物的影响。方法：构建27-nt miRNA 高表达质粒，并将其转染至HUVECs。MTT 法检测细胞的增殖情况，划痕实验检测细胞的迁移能力，Western Blot检测27-nt miRNA 及细胞eNOS 蛋白的表达情况，ELISA法检测eNOS活性，硝酸还原法测定细胞培养上清液中NO的含量。结果：27-nt miRNA 对 HUVECs 的增殖有强烈的抑制作用(0.674±0.093 vs 0.315±0.013, 0.743±0.076 vs 0.315±0.013, P < 0.05)；27-nt miRNA 对HUVECs 的迁移有显著的抑制作用(0.483±0.009 vs 0.806±0.017, 0.465±0.047 vs 0.806±0.017, P < 0.05)；27-nt miRNA 显著降低eNOS 蛋白的表达(0.410±0.004 vs 0.645±0.007, 0.483±0.009 vs 0.645±0.007，P < 0.05)及明显抑制eNOS的活性(1.093±0.357 vs 5.034±0.509, 1.707±0.652 vs 5.034±0.509，P＜0.05)；27-nt miRNA明显抑制NO的合成与释放(70.687±4.432 vs 136.803±6.913，75.264±4.481 vs 136.803±6.913，P＜0.05）。结论：27-nt miRNA 高表达明显抑制eNOS 基因的表达及活性；27-nt miRNA 明显抑制NO的合成和释放，可能成为血管性疾病治疗的分子靶点。 |
| 英文摘要: |
| ABSTRACT Objective: We previously reported 27-nt miRNA regulated the expression of eNOS. This study was to further investigate the role of 27-nt miRNA on eNOS gene expression, activity and the impact of its metabolites. Methods: The endothelial cells proliferation inhibition rate was detected by MTT assay. Migration ability of endothelial cells was measured by scratch wound model in vitro. The protein expression level of eNOS was detected by Western Blot. The eNOS activity was detected by ELISA. The content of metabolite NO in cell culture supernatant was detected by nitrate reduction. Results: Over-expression of 27-nt miRNA significantly inhibited the endothelial cells proliferation (0.674±0.093 vs 0.315±0.013, 0.743±0.076 vs 0.315±0.013, P <0.05). Over-expression of 27-nt miRNA significantly inhibited the migration of HUVEC (0.483±0.009 vs 0.806±0.017, 0.465±0.047 vs 0.806, P<0.05). Over-expression of 27-nt miRNA significantly decreased eNOS protein expression (0.410±0.004 vs 0.645±0.007, 0.483± 0.009 vs 0.645±0.007, P < 0.05) and significantly suppressed eNOS activity (1.093±0.357 vs 5.034±0.509, 1.707±0.652 vs 5.034±0.509, P<0.05). Over-expression of 27-nt miRNA significantly inhibited the synthesis and release of NO (70.687±4.432 vs 136.803±6.913, 75.264±4.481 vs 136.803±6.913, P<0.05). Conclusion: The 27nt-miRNA suppresses eNOS gene expression and activity in endothelial cells. 27-nt miRNA inhibits the synthesis and release of NO, the molecular target may be the treatment of vascular disease. |
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