title: Knockdown of Nrf2 inhibits the angiogenesis of rat cardiac micro-vascular endothelial cells under hypoxic conditions.
abstract: Angiogenesis plays an important role in myocardial repair after myocardial infarction (MI). Cardiac micro-vascular endothelial cells (CMECs) are important participants in myocardial angiogenesis processes. Recent studies have revealed that Nuclear factor-erythroid 2-related factor 2 (Nrf2), a master transcription factor of endogenous anti-oxidative defense systems, exerts cardio-protection in the cardiovascular system. However, the role of Nrf2 in the process of myocardial angiogenesis and corresponding mechanisms are not fully understood. Thus, the present study investigated the role of Nrf2 in the angiogenesis of rat CMECs to hypoxia. Trans-well assay, three-dimensional Matrigel assay were used to determine cell migration and vascular tube formation. Real-time RT-PCR, ELISA and Western blot were measured mRNA and protein expression. Here, we report that the mRNA and protein expression of Nrf2 and heme oxygenase-1(HO-1) were temporarily upregulated under hypoxic condition. Furthermore, knock down of Nrf2 significantly suppressed the migration and vascular tube formation of rat CMECs to hypoxia, Nrf2 knockdown also significantly decreased HO-1 and vascular endothelial growth factor (VEGF) expression at 48 h after transfection under hypoxic condition. Finally, transfection of CMECs with the Nrf2 over-expressing lentiviral vector upregulated HO-1 expression with a concomitant increase in cell migration and vascular tube formation induced by hypoxia, and this effect was greatly attenuated in the presence of ZnPP (a HO-1 inhibitor). Taken together, these results suggest that Nrf2 may mediate the angiogenesis of CMECs under hypoxic condition, and HO-1 is involved in regulating the angiogenesis of CMECs through Nrf2. Therefore, Nrf2 is a potent regulator of hypoxia-condition mediated angiogenesis in CMECs, which may provide a therapeutic strategy for myocardial repair after MI.
authors.affiliation: Department of Cardiology, Southwest Hospital, The Third Military Medical University, Chongqing City 400038, P. R. China.
authors.forename: Lihong
authors.initials: L
authors.lastname: Kuang
authors.forename: Jian
authors.initials: J
authors.lastname: Feng
authors.forename: Guoxiang
authors.initials: G
authors.lastname: He
authors.forename: Tao
authors.initials: T
authors.lastname: Jing
mesh_terms: D000818:Animals; D042783:Endothelial Cells / Q000502:physiology*; D055785:Gene Knockdown Techniques; D051547:Heme Oxygenase-1 / Q000096:biosynthesis; D000860:Hypoxia / Q000503:physiopathology*; D008297:Male; D051267:NF-E2-Related Factor 2 / Q000235:genetics* / Q000502:physiology*; D009389:Neovascularization, Pathologic / Q000503:physiopathology*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D015854:Up-Regulation; D042461:Vascular Endothelial Growth Factor A / Q000096:biosynthesis
chemical_list: D051267:NF-E2-Related Factor 2; C495637:Nfe2l2 protein, rat; D012333:RNA, Messenger; D042461:Vascular Endothelial Growth Factor A; D051547:Heme Oxygenase-1
keywords: Angiogenesis; Cardiac microvascular endothelial cell; Hypoxia.; NF-E2-related factor 2
references.citation: Trends Biochem Sci. 1997 Jul;22(7):251-6
references.pmid: 9255066
references.citation: Biochem Biophys Res Commun. 2010 Jul 23;398(2):315-20
references.pmid: 20599714
references.citation: J Gerontol A Biol Sci Med Sci. 2012 Aug;67(8):821-9
references.pmid: 22219515
references.citation: Cancer Chemother Pharmacol. 2012 Feb;69(2):485-94
references.pmid: 21842204
references.citation: Peptides. 2012 Jan;33(1):92-100
references.pmid: 22100225
references.citation: Cardiovasc Res. 2009 Jul 15;83(2):354-61
references.pmid: 19351736
references.citation: Physiol Genomics. 2010 May;41(3):306-14
references.pmid: 20197421
references.citation: Lab Invest. 1984 Dec;51(6):624-34
references.pmid: 6209468
references.citation: Microvasc Res. 1995 Jul;50(1):119-28
references.pmid: 7476573
references.citation: Exp Eye Res. 2010 Apr;90(4):493-500
references.pmid: 20064509
references.citation: Am J Physiol. 1993 Feb;264(2 Pt 2):H639-52
references.pmid: 8447476
references.citation: Cancer Res. 2011 Mar 15;71(6):2260-75
references.pmid: 21278237
references.citation: J Biol Chem. 2009 May 15;284(20):13291-5
references.pmid: 19182219
references.citation: Nat Med. 1999 May;5(5):491-2
references.pmid: 10229223
references.citation: Funct Integr Genomics. 2011 Mar;11(1):119-37
references.pmid: 20922447
references.citation: Cardiovasc Res. 2008 May 1;78(2):294-300
references.pmid: 18093990
references.citation: Nat Med. 2003 Jun;9(6):653-60
references.pmid: 12778163
references.citation: Annu Rev Pharmacol Toxicol. 2007;47:89-116
references.pmid: 16968214
references.citation: Prog Cardiovasc Dis. 1997 Jan-Feb;39(4):351-60
references.pmid: 9050820
references.citation: Arterioscler Thromb Vasc Biol. 2009 Nov;29(11):1843-50
references.pmid: 19592468
references.citation: Arterioscler Thromb Vasc Biol. 2010 May;30(5):1007-13
references.pmid: 20185790
references.citation: Expert Opin Ther Targets. 2009 Jul;13(7):785-94
references.pmid: 19530984
references.citation: Endocr Rev. 2004 Aug;25(4):581-611
references.pmid: 15294883
references.citation: Brain Res. 2009 Jul 28;1282:133-41
references.pmid: 19445907
references.citation: Free Radic Biol Med. 2011 Jun 15;50(12):1749-59
references.pmid: 21443946
references.citation: J Cell Biochem. 2011 Jun;112(6):1524-31
references.pmid: 21328610
references.citation: Arterioscler Thromb Vasc Biol. 2013 Apr;33(4):760-8
references.pmid: 23413426
references.citation: J Ethnopharmacol. 2010 Jul 20;130(2):222-30
references.pmid: 20447451
references.citation: Circ Res. 2007 Sep 28;101(7):723-33
references.pmid: 17673673
references.citation: Toxicol Appl Pharmacol. 2010 May 1;244(3):291-9
references.pmid: 20083128
references.citation: Eur Heart J. 2004 May;25(10):854-8
references.pmid: 15140533