M atrix metalloproteinase-2 mediates cytokine-induced myocardial contractile dysfunction (original) (raw)
Related papers
British Journal of Pharmacology, 2008
Background and purpose: The potent oxidant peroxynitrite (ONOO À) induces mechanical dysfunction in the intact heart in part through activation of matrix metalloproteinase-2 (MMP-2). This effect may be independent of the proteolytic actions of MMPs on extracellular matrix proteins. The purpose of this study was to examine the effects of ONOO À on contractile function at the level of the single cardiac myocyte and whether this includes the action of MMPs. Experimental approach: Freshly isolated ventricular myocytes from adult rats were superfused with Krebs-Henseleit buffer at 21 1C and paced at 0.5 Hz. Contractility was measured using a video edge-detector. ONOO À or decomposed ONOO À (vehicle control) were co-infused over 40 min to evaluate the contraction cease time (CCT). The effects of ONOO À on intracellular [Ca 2 þ ] were determined in myocytes loaded with calcium green-1 AM. MMP-2 activity was measured by gelatin zymography. Key results: ONOO À (30-600 mM) caused a concentration-dependent reduction in CCT. Myocytes subjected to 300 mM ONOO À had a shorter CCT than decomposed ONOO À (14.9 þ 1.5 vs 32.2 þ 3.5 min, n ¼ 7-8; Po0.05) and showed increased MMP-2 activity. The MMP inhibitors doxycycline (100 mM) or PD 166793 (2 mM) reduced the decline in CCT induced by 300 mM ONOO À. ONOO À caused shorter calcium transient cease time and significant alterations in intracellular [Ca 2 þ ] homoeostasis which were partially prevented by doxycycline. Conclusions and implications: This is the first demonstration that inhibition of MMPs protects the cardiac myocyte from ONOO À-induced contractile failure via an action unrelated to proteolysis of extracellular matrix proteins.
Journal of Molecular and Cellular Cardiology, 2006
The peroxynitrite-mediated activation of matrix metalloproteinase-2 (MMP-2) and subsequent cleavage of troponin I (TnI) in ventricular myocytes is a detrimental effect of ischemia/reperfusion injury. We hypothesized that acetaminophen, an effective antioxidant against peroxynitrite, would attenuate activation of MMP-2 and improve cardiac mechanical function. Isolated, perfused guinea pig hearts (Langendorff) were treated with either acetaminophen [0.35 mmol/l] or its vehicle and administered a bolus injection of peroxynitrite (6 μM) after reaching steady state function. Hemodynamic, metabolic, and mechanical effects were recorded, and coronary effluent concentrates or supernatant from heart homogenates were subjected to Western blotting and gelatin zymography. Hemodynamic and metabolic data showed no difference between acetaminophen-and vehicle-treated hearts. Mechanical data revealed that treatment with acetaminophen preserved contractile function (particularly diastolic function) after peroxynitrite administration. For example, 5 min after administration of peroxynitrite percent baseline -dP/dt max was 10 ± 3% and -4 ± 7% (P < 0.05) in acetaminophen-and vehicle-treated hearts, respectively. Western blotting and gel zymography revealed higher 72 kDa (pro-MMP-2) proteolytic activity in heart homogenates of vehicle-treated versus acetaminophen-treated hearts. In addition, Western blotting of heart homogenates showed increased degradative products of TnI in vehicle-treated versus acetaminophen-treated hearts. We conclude that acetaminophen is cardioprotective, at least in part, by attenuating peroxynitrite-activated, MMP-2-mediated cleavage of TnI.
Matrix Metalloproteinases in Myocardial Infarction and Heart Failure
Progress in molecular biology and translational science, 2017
Cardiovascular disease is the leading cause of death, accounting for 600,000 deaths each year in the United States. In addition, heart failure accounts for 37% of health care spending. Matrix metalloproteinases (MMPs) increase after myocardial infarction (MI) and correlate with left ventricular dysfunction in heart failure patients. MMPs regulate the remodeling process by facilitating extracellular matrix turnover and inflammatory signaling. Due to the critical role MMPs play during cardiac remodeling, there is a need to better understand the pathophysiological mechanism of MMPs, including the biological function of the downstream products of MMP proteolysis. Future studies developing new therapeutic targets that inhibit specific MMP actions to limit the development of heart failure post-MI are warranted. This chapter focuses on the role of MMPs post-MI, the efficiency of MMPs as biomarkers for MI or heart failure, and the future of MMPs and their cleavage products as targets for pr...
Objective: Tissue inhibitors of metalloproteinases (TIMPs) are downregulated in the failing human heart. The objective of the present study was to test the hypothesis that cytokines might be involved in the regulation of TIMPs in cardiac cells. Methods: Neonatal Sprague-Dawley rat ventricular cells were exposed to 100 units / ml tumor necrosis factor-a and / or 5 ng / ml interleukin-1b. The mRNA and protein expression of TIMPs-1-4 and disintegrin metalloproteinase was analyzed using Northern blot, ELISA and / or Western blot, respectively. Proteolytic activity and extracellular matrix degradation and turnover were determined using gelatin zymography and pulse-chase experiments. Results: The TIMP-1 mRNA was upregulated in cardiac cells, while TIMP-1 protein levels were unchanged in myocytes but downregulated in non-myocytes. The TIMP-2 expression did not change with the cytokine treatment. TIMP-3 was downregulated at both the mRNA and protein levels in cardiac cells. TIMP-4 protein was transiently increased and then returned to control level. In contrast, disintegrin metalloproteinase mRNA and protein were significantly upregulated in those cells. The gelatinolytic activity and extracellular matrix protein degradation were significantly increased. Conclusions: Tumor necrosis factor-a and interleukin-1b regulate the expression of TIMPs and disintegrin metalloproteinase, which may in turn contribute to the increased matrix degradation in cardiac cells. Since heart failure in humans is characterized by both re-expression of myocardial cytokines and remodeling of the extracellular matrix, those in vitro results suggest a potential role for those cytokines in the regulation of extracellular matrix remodeling and therefore in the transition to the end-stage heart failure phenotype.
AJP: Heart and Circulatory Physiology, 2010
We have previously shown that the inhibition of myocardial nitric oxide (NO) and peroxynitrite-matrix metalloproteinase (MMP) signaling by early preconditioning (PC) is involved in its cardioprotective effect. Therefore, in the present study, we investigated the role of NO and peroxynitrite-MMP signaling in the development of late PC. PC was performed by five consecutive cycles of 4-min coronary occlusion and 4-min reperfusion in anesthetized rats in vivo. Twenty-four hours later, hearts were subjected to a 30-min coronary occlusion followed by 180-min reperfusion to measure infarct size. In separate experiments, heart tissue was sampled to measure biochemical parameters before and 3, 6, 12, or 24 h after the PC protocol, respectively. Late PC decreased infarct size, increased cardiac inducible NO synthase (iNOS) activity and gene expression, and decreased SOD activity at 24 h significantly compared with sham-operated controls. Late PC increased cardiac superoxide levels significant...
Journal of Biological Chemistry, 2010
The membrane type-1 matrix metalloproteinase (MT1-MMP) is a unique member of the MMP family, but induction patterns and consequences of MT1-MMP overexpression (MT1-MMPexp), in a left ventricular (LV) remodeling process such as myocardial infarction (MI), have not been explored. MT1-MMP promoter activity (murine luciferase reporter) increased 20-fold at 3 days and 50-fold at 14 days post-MI. MI was then induced in mice with cardiac restricted MT1-MMPexp (n ؍ 58) and wild type (WT, n ؍ 60). Post-MI survival was reduced (67% versus 46%, p < 0.05), and LV ejection fraction was lower in the post-MI MT1-MMPexp mice compared with WT (41 ؎ 2 versus 32 ؎ 2%,p < 0.05). In the post-MI MT1-MMPexp mice, LV myocardial MMP activity, as assessed by radiotracer uptake, and MT1-MMP-specific proteolytic activity using a specific fluorogenic assay were both increased by 2-fold. LV collagen content was increased by nearly 2-fold in the post-MI MT1-MMPexp compared with WT. Using a validated fluorogenic construct, it was discovered that MT1-MMP proteolytically processed the pro-fibrotic molecule, . 2 The abbreviations used are: MMP, matrix metalloproteinase; LV, left ventricular; MI, myocardial infarction; LTBP-1, latency-associated transforming growth factor-1 binding protein; %ID, percentage of injected dose; MOPS, 4-morpholinepropanesulfonic acid; ANOVA, analysis of variance; rtPCR, real-time PCR. Downloaded from FIGURE 3. Representative LV sections in which MT1-MMP was localized by immunofluorescence using confocal microscopy in WT and MT1-MMPexp mice. Green fluorescence represents MT1-MMP, red fluorescence is indicative of rhodamine-phalloidin, and the blue stain is indicative of nuclei. Increased relative abundance and staining intensity for MT1-MMP was observed along the myocyte-matrix interface in LV sections taken MT1-MMPexp mice. At 14 days post-MI, a robust increase in MT1-MMP staining was observed within the MI region for both WT and MT1-MMPexp mice. However, in the MT1-MMPexp mice, a more robust and greater staining pattern was clearly observed within the remote regions. (Scale ϭ 20 m.)
Diabetes, 2007
We investigated the effect of the angiotensin type 1 (AT-1) receptor antagonist, irbesartan, on matrix metalloproteinase (MMP) activity and cardiac cytokines in an animal model of diabetic cardiomyopathy. Diabetes was induced in 20 C57/bl6 mice by injection of streptozotocin (STZ). These animals were treated with irbesartan or placebo and were compared with nondiabetic controls. Left ventricular (LV) function was measured by pressure-volume loops with parameters for systolic function (end systolic elastance [Ees]) and diastolic function (cardiac stiffness) 8 weeks after STZ treatment. The cardiac protein content of interleukin (IL)1 and transforming growth factor (TGF)1 were measured by enzyme-linked immunosorbent assay. The total cardiac collagen content and collagen type 1 and 3 were measured by histochemestry, and MMP-2 activity was measured by gelatin zymography. LV dysfunction was documented by impaired Ees and diastolic stiffness in STZ mice compared with controls. This was accompanied by increased TGF, IL1, and fibrosis and decreased MMP-2 activity. Treatment with irbesartan attenuated LV dysfunction, IL1, TGF, and cardiac fibrosis compared with untreated diabetic animals and normalized MMP activity. These findings present evidence that AT-1 receptor antagonists attenuate cardiac failure by decreasing cardiac inflammation and normalizing MMP activity, leading to normalized cardiac fibrosis in STZ-induced diabetic cardiomyopathy. Diabetes 56:641-646, 2007 RESEARCH DESIGN AND METHODS Diabetes was induced in 20 C57i/BL6J mice by injection of STZ (50 mg/kg i.p. for 5 days) dissolved in 0.1 mol/l sodium citrate, pH 4.5, as previously described (11). This dose is known to induce no acute renal failure in C57i/BL6J mice (12). Hyperglycemia (glucose Ͼ22 mmol/l) was confirmed 7 days later using a reflectance meter (Acutrend; Boehringer, Mannheim,
The role of matrix metalloproteinases in heart disease
Cardiovascular Research, 1996
lar matrix also contains hetero-polysaccharides (glycosaminoglycans), glycoproteins (heparan sulphate and chondroitin sulphate proteoglycans), microfibrillar proteins (fibrillin and fibulin) and elastin.
Circulation, 2010
Background— Titin is the largest mammalian (≈3000 to 4000 kDa) and myofilament protein that acts as a molecular spring in the cardiac sarcomere and determines systolic and diastolic function. Loss of titin in ischemic hearts has been reported, but the mechanism of titin degradation is not well understood. Matrix metalloproteinase-2 (MMP-2) is localized to the cardiac sarcomere and, on activation in ischemia/reperfusion injury, proteolyzes specific myofilament proteins. Here we determine whether titin is an intracellular substrate for MMP-2 and if its degradation during ischemia/reperfusion contributes to cardiac contractile dysfunction. Methods and Results— Immunohistochemistry and confocal microscopy in rat and human hearts showed discrete colocalization between MMP-2 and titin in the Z-disk region of titin and that MMP-2 is localized mainly to titin near the Z disk of the cardiac sarcomere. Both purified titin and titin in skinned cardiomyocytes were proteolyzed when incubated wit...