Interleukin-1beta increases expression and activity of matrix metalloproteinase-2 in cardiac microvascular endothelial cells: role of PKCalpha/beta1 and MAPKs.
Matrix metalloproteinases (MMPs), a family of extracellular endopeptidases, are implicated in angiogenesis because of their ability to selectively degrade components of the extracellular matrix. Interleukin-1beta (IL-1beta), increased in the heart post-myocardial infarction (post-MI), plays a protective role in the pathophysiology of left ventricular (LV) remodeling following MI. Here we studied expression of various angiogenic genes affected by IL-1beta in cardiac microvascular endothelial cells (CMECs) and investigated the signaling pathways involved in the regulation of MMP-2. cDNA array analysis of 96 angiogenesis-related genes indicated that IL-1beta modulates the expression of numerous genes, notably increasing the expression of MMP-2, not MMP-9. RT-PCR and Western blot analyses confirmed increased expression of MMP-2 in response to IL-1beta. Gelatin in-gel zymography and Biotrak activity assay demonstrated that IL-1beta increases MMP-2 activity in the conditioned media. IL-1beta activated ERK1/2, JNKs, and protein kinase C (PKC), specifically PKCalpha/beta(1), and inhibition of these cascades partially inhibited IL-1beta-stimulated increases in MMP-2. Inhibition of PKCalpha/beta(1) failed to inhibit ERK1/2. However, concurrent inhibition of PKCalpha/beta(1) and ERK1/2 almost completely inhibited IL-1beta-mediated increases in MMP-2 expression. Inhibition of p38 kinase and nuclear factor-kappaB (NF-kappaB) had no effect. Pretreatment with superoxide dismutase (SOD) mimetic, MnTMPyP, increased MMP-2 protein levels, whereas pretreatment with SOD and catalase mimetic, EUK134, partially inhibited IL-1beta-stimulated increases in MMP-2 protein levels. Exogenous H(2)O(2) significantly increased MMP-2 protein levels, whereas superoxide generation by xanthine/xanthine oxidase had no effect. This in vitro study suggests that IL-1beta modulates expression and activity of MMP-2 in CMECs.
Mountain
Deidra J H
DJ
Dept. of Physiology, James H. Quillen College of Medicine, East Tennessee State Univ., PO Box 70576, Johnson City, TN 37614, USA. singhk@etsu.edu
Singh
Mahipal
M
Menon
Bindu
B
Singh
Krishna
K
D000818:Animals; D002374:Catalase / Q000494:pharmacology; D002478:Cells, Cultured; D003331:Coronary Vessels / Q000166:cytology*; D042783:Endothelial Cells / Q000502:physiology*; D004789:Enzyme Activation; D053583:Interleukin-1beta / Q000502:physiology*; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000378:metabolism; D020935:MAP Kinase Signaling System / Q000502:physiology*; D008297:Male; D020778:Matrix Metalloproteinase 2 / Q000096:biosynthesis / Q000378:metabolism*; D020780:Matrix Metalloproteinase 9 / Q000502:physiology; D008665:Metalloporphyrins / Q000494:pharmacology; D008833:Microcirculation / Q000166:cytology; D009942:Organometallic Compounds / Q000494:pharmacology; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism*; D064546:Protein Kinase C beta; D051571:Protein Kinase C-alpha / Q000037:antagonists & inhibitors / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012459:Salicylates / Q000494:pharmacology; D013482:Superoxide Dismutase / Q000494:pharmacology
C433928:EUK-134; D053583:Interleukin-1beta; D007527:Isoenzymes; D008665:Metalloporphyrins; C502230:Mn(III) 5,10,15,20-tetrakis(N-methylpyridinium-2-yl)porphyrin; D009942:Organometallic Compounds; D012459:Salicylates; D002374:Catalase; D013482:Superoxide Dismutase; D011493:Protein Kinase C; D064546:Protein Kinase C beta; D051571:Protein Kinase C-alpha; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9