Effects of losartan on expression of matrix metalloproteinase-2,JNK1/2 and proliferation in cardiac fibroblast

Abstract

Abstract: AIM: To elucidate the effects of losartan on the expression of matrix metalloproteinases-2, JNK1/2 and proliferation in cardiac fibroblast. METHODS: Neonatal rat cardiac fibroblasts were cultured.The cells proliferation was determined by MTT.To determine effects of Ang Ⅱ on JNK1/2 activity, cells were incubated (for 0,2, 5, 10, 30, 60, 120 min) in serum-free media with Ang Ⅱ, and the other group fibroblasts were exposed to serum-free media with or without Ang Ⅱ and losartan (Ang Ⅱ 100 nmol/L, Ang Ⅱ 100 nmol/L +losartan 100 nmol/L, losartan100 nmol/L, losartan for 45 min before).Cells protein was collected with MBST buffer.The relative abundance of MMP-2, JNK1/2 and p-JNK1/2 in cells was determined by immunoblotting.The secretion of MMP-2 in media of cell culture was determined by ELISA. RESULTS: Ang Ⅱ increased the proliferation of CFB in a dose-dependent manner, whereas losartan decreased the proliferation of CFB stimulated by Ang Ⅱ in a dose-dependant manner, too (P<0.05).The relative abundance of JNK1/2 was highest in Ang Ⅱ of the 2-minstimulated group.Ang Ⅱ increased expression of JNK1/2 andMMP-2 protein (P<0.05), on the contrary, losartan inhibited JNK1/2 and MMP-2 protein expression. CONCLUSION: Ang Ⅱ induce the increase of proliferation of CFB, expression of JNK1/2 and MMP-2 in CFB, and losartan inhibits these effects of Ang Ⅱ.

Key words: matrix metalloproteinase-2, JNK1/2, losartan, cardiac fibroblasts

CLC Number:

R969

XIAO Yun-bin, QIN Xu-ping, QIN Li, LIAO Duan-fang, HUANG Hong-lin. Effects of losartan on expression of matrix metalloproteinase-2,JNK1/2 and proliferation in cardiac fibroblast[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2007, 12(1): 72-77.

References

[1] Cohn JN, Ferral R, Sharpe N. Cardiac remodeling concepts and clinical implications: a consensus paper from an international forum of cardiac remodeling[J]. J Am Coll Cardiol, 2000;35: 562-582.
[2] Sun Y, Weber KT. Angiotensin converting enzyme and myofibroblasts during tissue repair in the rat heart[J]. J Mol Cell Cardiol, 1996;28: 851-858.
[3] Dollery CM, Mc Ewan JR, Henney AM. Matrix metalloproteinases and cardiovascular disease[J]. Circ Res, 1995;77: 863-868.
[4] Spinale FG, Coker ML, Heung/LJ, et al. A matrix metalloproteinase induction activation system exists in the human left ventricular myocardium and is upregulated in heart failure[J]. Circulation, 2000;102: 1944-1949.
[5] Iwanaga Y, Aoyama T, Kihara Y, et al. Excessive activation of matrix metalloproteinases coincides with left ventricular remodeling during transition from hypertrophy to heart failure in hypertensive rats[J]. J Am Coll Cardiol, 2002;39: 1384-1391.
[6] Reinhardh D, Sigusch HH, Henbe J, et al. Cardiac remodeling in end stage heart failure: upregulation of matrix metalloproteinase(MMP) irrespective of the underlying disease, and evidence for a direct inhibitory effect of ACE inhibitors on MMP[J]. Heart, 2002;88: 525-530.
[7] Chancey AL, Brower GL, Peterson JT, et al. Effects of matrix metallproteinase inhibition on ventricular remodeling due to volume overload[J]. Circulation, 2002;105: 1983-1988.
[8] Swynghedauw B. Molecular mechanisms of myocardial remodeling[J]. Physiol Rev, 1999;79: 215-262.
[9] Lijnen P, Petrov V. Renin-angiotensin sy stem, hypertrophy and gene expression in cardiac myocytes[J]. J Mol Cell Cardial, 1999;31: 949-970.
[10] Lijnen PJ, Petrov VV. Role of intracardiac rennin-angiotensinaldosterone system in extracellular matrix remodeling[J]. Methods Find Exp Clin Pharmacol, 2003;25: 541-564.
[11] Gonzalez A, Lopez B, Querejeta R, et al. Regulation of myocardial fibrillar collagen by angiotensinⅡ: A role in hypertensive heart disease[J]. J Mol Cell Cardial, 2002;34: 1585-1593.
[12] Yamazaki T, Komuro I, Kudoh S, et al. Angiotensin Ⅱ partly mediates mechanical stress-induced cardiac hypertrophy[J]. Circ Res, 1995;77: 258-265.
[13] Kudo S, Komuro I, Mizuno T, et al. Angiotensin Ⅱ stimulates c-Jun NH2-terminal kinase in cultured cardiac myocytes of neonatal rats[J]. Circ Res, 1997;80: 139-146.
[14] Gennaro G, Menard C, Michaud SE, et al. Inhibition of vascular smooth muscle cell proliferation and neointimal formation in injured arteries by a novel, oral mitogen-activated protein kinase extracellular signal-regulated kinase inhibitor[J]. Circulation, 2004;110: 3367-3371.
[15] Izumi Y, Kim S, Zhan Y, et al. Important role of angiotensin Ⅱ-mediated c-Jun NH2-Terminal Kinasa actiation in cardiac hypertrophy in hypertensive rats[J]. Hypertension, 2000;36: 511-516.
[16] Gurantz D, Cowling RT, Villarreal FJ, et al. Tumor necrosis factor-a upregulates angiotensin Ⅱtype 1 receptors on cardiac fibroblast[J]. Circ Res, 1999;85: 272-279.
[17] Ahmed SH, Clard LL, PenningtonWR, et al. Matrix metalloproteinases tissue inhibitors of metalloproteinases. relationship between changes in proteolytic determinants of matrix composition and structural, functional, and clinical manifestations of hypertensive heart disease[J]. Circulation, 2006;24: 1156-1170.
[18] Gradman AH, Alfayoumi F. From left ventricular hypertrophy to congestive heart failure: management of hypertensive heart disease[J]. Prog Cardiovasc Dis, 2006;48: 326-341.
[19] Ferrari R. Perindopril and remodeling in elderly with acute myocardial infarction investigators: Effects of angiotensin-converting enzyme inhibition with perindopril on left ventricular remodeling and clinical outcome: results of the randomized perindopril and remodeling in elderly with acute myocardial infarction study[J]. Arch InternMed, 2006;166: 659-566.
[20] Hattori Y, Akimoto K, Nishikimi T, et al. Activation of AMPactivated protein kinase enhances angiotensin ii-induced proliferation in cardiac fibroblasts[J]. Hypertension, 2006;47: 265-270.
[21] Qin XP, Ye F, Liao DF, et al. Involvement of calcitonin gene-related peptide in the depressor effects of losartan and perindopril in rats[J]. European J Pharmacol, 2003;464: 63-67.
[22] Xiao YB, Huang HL, Qin XP, et al. Effects of losartan on the expression of matrix metalloproteinases-2, collagen Ⅲ and JNK1/2 in the myocardium of hypertensive rats[J]. Chin J Hypertension, 2004;6: 551-555.
[23] Nicoletti A, Heudes D, Hinglais N. Left ventricular fibrosis in renovascular hypertensive rats: effect of losartan and spironolactone[J]. Hypertension, 1995;26: 101-111.
[24] KarinM. The regulation of AP-1 activity by mitogen-activated protein kinases[J]. J Biol Chem, 1995;270: 16483-16486.
[25] Dollery CM, McEwan JR, Henney AM. Matrix metalloproteinases and cardiovascular disease[J]. Circ Res, 1995;77: 863-868.