title: Metformin inhibits angiotensin II-induced differentiation of cardiac fibroblasts into myofibroblasts.
abstract: Differentiation of cardiac fibroblasts into myofibroblasts is a critical event in the progression of cardiac fibrosis that leads to pathological cardiac remodeling. Metformin, an antidiabetic agent, exhibits a number of cardioprotective properties. However, much less is known regarding the effect of metformin on cardiac fibroblast differentiation. Thus, in the present study, we examined the effect of metformin on angiotensin (Ang) II-induced differentiation of cardiac fibroblasts into myofibroblasts and its underlying mechanism. Adult rat cardiac fibroblasts were stimulated with Ang II (100 nM) in the presence or absence of metformin (10-200 µM). Ang II stimulation induced the differentiation of cardiac fibroblasts into myofibroblasts, as indicated by increased expression of α-smooth muscle actin (α-SMA) and collagen types I and III, and this effect of Ang II was inhibited by pretreatment of cardiac fibroblasts with metformin. Metformin also decreased Ang II-induced reactive oxygen species (ROS) generation in cardiac fibroblasts via inhibiting the activation of the PKC-NADPH oxidase pathway. Further experiments using PKC inhibitor calphostin C and NADPH oxidase inhibitor apocynin confirmed that inhibition of the PKC-NADPH oxidase pathway markedly attenuated Ang II-induced ROS generation and myofibroblast differentiation. These data indicate that metformin inhibits Ang II-induced myofibroblast differentiation by suppressing ROS generation via the inhibition of the PKC-NADPH oxidase pathway in adult rat cardiac fibroblasts. Our results provide new mechanistic insights regarding the cardioprotective effects of metformin and provide an efficient therapeutic strategy to attenuate cardiac fibrosis.
authors.affiliation: Department of Cardiology, Drum Tower Hospital, Nanjing University Medical School, Nanjing, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Bai
authors.forename: Na
authors.initials: N
authors.lastname: Zhang
authors.forename: Ying
authors.initials: Y
authors.lastname: Hua
authors.forename: Bingjian
authors.initials: B
authors.lastname: Wang
authors.forename: Lin
authors.initials: L
authors.lastname: Ling
authors.forename: Albert
authors.initials: A
authors.lastname: Ferro
authors.forename: Biao
authors.initials: B
authors.lastname: Xu
mesh_terms: D000804:Angiotensin II / Q000494:pharmacology*; D000818:Animals; D015153:Blotting, Western; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D005347:Fibroblasts / Q000166:cytology* / Q000187:drug effects; D008297:Male; D008687:Metformin / Q000494:pharmacology*; D058628:Myofibroblasts / Q000166:cytology* / Q000187:drug effects; D019255:NADPH Oxidases / Q000378:metabolism; D051381:Rats; D017382:Reactive Oxygen Species / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D017382:Reactive Oxygen Species; D000804:Angiotensin II; D008687:Metformin; D019255:NADPH Oxidases
references.citation: Pharmacol Ther. 2009 Aug;123(2):255-78
references.pmid: 19460403
references.citation: Hypertension. 2004 Jun;43(6):1195-201
references.pmid: 15123578
references.citation: Diabetes. 2008 Feb;57(2):451-9
references.pmid: 17977949
references.citation: Arterioscler Thromb Vasc Biol. 2008 May;28(5):863-70
references.pmid: 18309113
references.citation: J Cardiovasc Pharmacol. 2011 Apr;57(4):380-8
references.pmid: 21242811
references.citation: Am J Physiol Heart Circ Physiol. 2008 Jun;294(6):H2879-88
references.pmid: 18441197
references.citation: Kidney Int. 2011 May;79(9):944-56
references.pmid: 21307839
references.citation: Circ Res. 2002 Sep 6;91(5):406-13
references.pmid: 12215489
references.citation: Hepatology. 2006 Nov;44(5):1267-77
references.pmid: 17058265
references.citation: Eur J Pharmacol. 2009 Aug 15;616(1-3):155-9
references.pmid: 19470381
references.citation: Am J Physiol Lung Cell Mol Physiol. 2000 Dec;279(6):L1005-28
references.pmid: 11076791
references.citation: Am J Physiol Heart Circ Physiol. 2006 Jan;290(1):H323-30
references.pmid: 16143656
references.citation: Hypertension. 2007 Feb;49(2):241-8
references.pmid: 17190878
references.citation: J Am Coll Cardiol. 2003 Nov 19;42(10):1845-54
references.pmid: 14642698
references.citation: Am J Physiol Heart Circ Physiol. 2006 Oct;291(4):H1738-45
references.pmid: 16714359
references.citation: J Hypertens. 2004 Jan;22(1):47-50
references.pmid: 15106793
references.citation: Am J Physiol Cell Physiol. 2007 Jan;292(1):C82-97
references.pmid: 16870827
references.citation: Respir Res. 2005 Jan 21;6:11
references.pmid: 15663794
references.citation: Methods Find Exp Clin Pharmacol. 2000 Dec;22(10):709-23
references.pmid: 11346891
references.citation: Cell Mol Life Sci. 2008 Sep;65(18):2924-32
references.pmid: 18726071
references.citation: Circ Res. 2005 Oct 28;97(9):900-7
references.pmid: 16179589
references.citation: Cardiovasc Res. 2010 Aug 1;87(3):504-13
references.pmid: 20200042
references.citation: Circ Res. 2001 Oct 12;89(8):661-9
references.pmid: 11597988
references.citation: Am J Physiol Heart Circ Physiol. 2011 Aug;301(2):H459-68
references.pmid: 21572014
references.citation: Arterioscler Thromb Vasc Biol. 2007 Dec;27(12):2627-33
references.pmid: 17916771
references.citation: Hypertension. 2003 Dec;42(6):1075-81
references.pmid: 14581295
references.citation: Cardiovasc Res. 2005 Jul 1;67(1):50-9
references.pmid: 15949469
references.citation: Circulation. 2009 May 19;119(19):2568-77
references.pmid: 19414638
references.citation: Lancet. 1998 Sep 12;352(9131):854-65
references.pmid: 9742977
references.citation: Basic Res Cardiol. 1993;88 Suppl 1:183-201
references.pmid: 8357333
references.citation: Hypertension. 2002 Feb;39(2):258-63
references.pmid: 11847194
references.citation: Free Radic Biol Med. 2009 Nov 1;47(9):1239-53
references.pmid: 19628035
references.citation: Cardiovasc Res. 2007 Aug 1;75(3):519-29
references.pmid: 17543901
references.citation: Free Radic Biol Med. 2000 May 15;28(10):1456-62
references.pmid: 10927169
references.citation: Am J Physiol Cell Physiol. 2009 Apr;296(4):C693-700
references.pmid: 19193863
references.citation: Proc Natl Acad Sci U S A. 2005 Jan 11;102(2):437-42
references.pmid: 15625103
references.citation: Metabolism. 2009 Apr;58(4):525-33
references.pmid: 19303974
references.citation: Biochem Pharmacol. 2006 Jul 14;72(2):176-83
references.pmid: 16730666
references.citation: Br J Pharmacol. 2010 Jul;160(6):1273-92
references.pmid: 20590619
references.citation: Am J Physiol Heart Circ Physiol. 2005 Mar;288(3):H1131-8
references.pmid: 15498824
references.citation: Mol Nutr Food Res. 2011 Sep;55 Suppl 2:S227-40
references.pmid: 21812107