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1.
Clin Biochem ; 73: 70-76, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31386834

RESUMO

BACKGROUND: Chromosomal region 9p21.3 is most robustly associated with coronary artery disease (CAD) in western European populations. However, heterogeneity in CAD phenotypes leads to uncertainty whether 9p21.3 is associated with stable and/or acute clinical presentations of CAD. 9p21.3 is rich in regulatory elements, but the underlying mechanisms of its actions in CAD remain unclear. We investigate the association of 9p21.3 two haplotype blocks lead variants (rs10757278 and rs518394) with first-ever non-fatal myocardial infarction (MI) in CAD patients and their association with CDKN2B mRNA expression in peripheral blood mononuclear cells 6 months after the event. METHODS: We included CAD patients with sustained first MI (n = 523) and controls (n = 583). Gene expression was assessed in 72 patients 6 months after MI and 43 healthy controls. TaqMan® technology was used for the gene expression and genotyping analysis. RESULTS: CDKN2B mRNA was significantly lower in MI patients compared with the controls (p = 0.002) and in patients carrying the rs10757278 G risk allele versus AA homozygotes (p = 0.012) 6 months after the event. While we confirmed the association of rs10757278 with CDKN2B expression in MI patients, we failed to find an association between the investigated variants and MI or disease burden. CONCLUSIONS: We suggest a dysregulation of gene expression in the 9p21.3 region six months after acute MI, which is affected by a genetic variant in patients. The rs10757278 rare allele is one factor that might lead to prolonged risk for proatherogenic complications.


Assuntos
Cromossomos Humanos Par 9/genética , Doença da Artéria Coronariana , Inibidor de Quinase Dependente de Ciclina p15/biossíntese , Regulação da Expressão Gênica , Haplótipos , Infarto do Miocárdio , Elementos de Resposta , Adulto , Idoso , Alelos , Cromossomos Humanos Par 9/metabolismo , Doença da Artéria Coronariana/enzimologia , Doença da Artéria Coronariana/genética , Inibidor de Quinase Dependente de Ciclina p15/genética , Feminino , Seguimentos , Homozigoto , Humanos , Masculino , Pessoa de Meia-Idade , Infarto do Miocárdio/enzimologia , Infarto do Miocárdio/genética
2.
Adv Mater ; 31(40): e1902900, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31408234

RESUMO

Although in situ restoration of blood supply to the infarction region and attenuating pre-existing extracellular matrix degradation remain potential therapeutic approaches for myocardial infarction (MI), local delivery of therapeutics has been limited by low accumulation (inefficacy) and unnecessary diffusion (toxicity). Here, a dual functional MI-responsive hydrogel is fabricated for on-demand drug delivery to promote angiogenesis and inhibit cardiac remodeling by targeting upregulated matrix metalloproteinase-2/9 (MMP-2/9) after MI. A glutathione (GSH)-modified collagen hydrogel (collagen-GSH) is prepared by conjugating collagen amine groups with GSH sulfhydryl groups and the recombinant protein GST-TIMP-bFGF (bFGF: basic fibroblast growth factor) by fusing bFGF with glutathione-S-transferase (GST) and MMP-2/9 cleavable peptide PLGLAG (TIMP). Specific binding between GST and GSH significantly improves the amount of GST-TIMP-bFGF loaded in collagen-GSH hydrogel. The TIMP peptide enclosed between GST and bFGF responds to MMPs for on-demand release during MI. Additionally, the TIMP peptide is a competitive substrate of MMPs that inhibits the excessive degradation of cardiac matrix by MMPs after MI. GST-TIMP-bFGF/collagen-GSH hydrogels promote the recovery of MI rats by enhancing vascularization and ameliorating myocardium remodeling. The results suggest that on-demand growth factor delivery by synchronously controlling binding and responsive release to promote angiogenesis and attenuate cardiac remodeling might be promising for the treatment of ischemic heart disease.


Assuntos
Portadores de Fármacos/química , Fator 2 de Crescimento de Fibroblastos/química , Hidrogéis/química , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/metabolismo , Infarto do Miocárdio/tratamento farmacológico , Células 3T3 , Animais , Colágeno/química , Liberação Controlada de Fármacos , Fator 2 de Crescimento de Fibroblastos/farmacologia , Fator 2 de Crescimento de Fibroblastos/uso terapêutico , Glutationa/química , Camundongos , Infarto do Miocárdio/enzimologia , Infarto do Miocárdio/fisiopatologia , Neovascularização Fisiológica/efeitos dos fármacos , Ratos , Regulação para Cima/efeitos dos fármacos
3.
PLoS One ; 14(5): e0216385, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31048932

RESUMO

FINDINGS: Here, we demonstrate that OP2113 (5-(4-Methoxyphenyl)-3H-1,2-dithiole-3-thione, CAS 532-11-6), synthesized and used as a drug since 1696, does not act as an unspecific antioxidant molecule (i.e., as a radical scavenger) but unexpectedly decreases mitochondrial reactive oxygen species (ROS/H2O2) production by acting as a specific inhibitor of ROS production at the IQ site of complex I of the mitochondrial respiratory chain. Studies performed on isolated rat heart mitochondria also showed that OP2113 does not affect oxidative phosphorylation driven by complex I or complex II substrates. We assessed the effect of OP2113 on an infarct model of ex vivo rat heart in which mitochondrial ROS production is highly involved and showed that OP2113 protects heart tissue as well as the recovery of heart contractile activity. CONCLUSION / SIGNIFICANCE: This work represents the first demonstration of a drug authorized for use in humans that can prevent mitochondria from producing ROS/H2O2. OP2113 therefore appears to be a member of the new class of mitochondrial ROS blockers (S1QELs) and could protect mitochondrial function in numerous diseases in which ROS-induced mitochondrial dysfunction occurs. These applications include but are not limited to aging, Parkinson's and Alzheimer's diseases, cardiac atrial fibrillation, and ischemia-reperfusion injury.


Assuntos
Complexo I de Transporte de Elétrons/metabolismo , Depuradores de Radicais Livres/farmacologia , Mitocôndrias Cardíacas/enzimologia , Infarto do Miocárdio/tratamento farmacológico , Espécies Reativas de Oxigênio/metabolismo , Animais , Modelos Animais de Doenças , Masculino , Mitocôndrias Cardíacas/patologia , Contração Miocárdica/efeitos dos fármacos , Infarto do Miocárdio/enzimologia , Infarto do Miocárdio/patologia , Fosforilação Oxidativa/efeitos dos fármacos , Ratos , Ratos Wistar
4.
Chem Commun (Camb) ; 55(44): 6193-6196, 2019 May 28.
Artigo em Inglês | MEDLINE | ID: mdl-31070620

RESUMO

Hydrogen sulfide (H2S) is an important signaling molecule with promising protective effects in many physiological and pathological processes. However, the study of H2S has been impeded by the lack of appropriate H2S donors that could mimic its slow-releasing process in vivo. Herein, we report the rational design, synthesis, and biological evaluation of a series of thioester-based H2S donors. These cysteine-activated H2S donors release H2S in a slow and controllable manner. Most of the donors comprising an allyl moiety showed significant cytoprotective effects in H9c2 cellular models of oxidative damage. The most potent donor 5e decreased the mitochondrial membrane potential (MMP) loss and lactate dehydrogenase (LDH) release in H2O2-stimulated H9c2 cells. More importantly, donor 5e exhibited a potent cardioprotective effect in an in vivo myocardial infarction (MI) mouse model by reducing myocardial infarct size and cardiomyocyte apoptosis. Taken together, our studies demonstrated that these new allyl thioesters are potential cardioprotective agents by releasing H2S.


Assuntos
Cardiotônicos/farmacologia , Cardiotônicos/uso terapêutico , Ésteres/química , Sulfeto de Hidrogênio/química , Compostos de Sulfidrila/química , Animais , Linhagem Celular , Modelos Animais de Doenças , Peróxido de Hidrogênio/farmacologia , L-Lactato Desidrogenase/metabolismo , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Camundongos , Infarto do Miocárdio/tratamento farmacológico , Infarto do Miocárdio/enzimologia , Infarto do Miocárdio/metabolismo , Estresse Oxidativo
5.
Molecules ; 24(4)2019 Feb 22.
Artigo em Inglês | MEDLINE | ID: mdl-30813328

RESUMO

A procedure to measure the serum concentration of glycogen phosphorylase during acute myocardial infarction is presented. This method was based on the synthesis of photoaffinity probes, and used the semiquantitative protein electrophoretic mobility shift technique. Three novel photoaffinity probes bearing different secondary tags were synthesized. Their potency was evaluated in an enzyme inhibition assay against rabbit muscle glycogen phosphorylase a (RMGPa). The inhibitory activity of probe 1 was only 100-fold less potent than the mother compound CP-320626. The photoaffinity labeling experiments were also performed, and a protein with molecular weight (MW) of about 90⁻100 kDa, which was consistent with the MW of GP, was clearly labeled by probe 1. A semiquantitative evaluation of the GP level in serum with probe 1 was also performed. The results showed that the protein band with a MW of about 90⁻100 kDa was tagged, and the concentration of the protein in serum was found to be between 25 and 50 ng/mL. Mass spectrometric analysis revealed that alpha-1,4 glucan phosphorylase (GPMM) was well-preserved in the bands.


Assuntos
Corantes Fluorescentes/química , Glicogênio Fosforilase Muscular/sangue , Infarto do Miocárdio/enzimologia , Marcadores de Fotoafinidade/química , Amidas/farmacologia , Animais , Química Click , Modelos Animais de Doenças , Ensaio de Desvio de Mobilidade Eletroforética , Glicogênio Fosforilase Muscular/química , Indóis/farmacologia , Masculino , Espectrometria de Massas , Estrutura Molecular , Peso Molecular , Infarto do Miocárdio/sangue , Coelhos
6.
Cardiovasc Drugs Ther ; 33(1): 13-23, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30637549

RESUMO

PURPOSE: Necroptosis is an important form of cell death following myocardial ischemia/reperfusion (I/R) and phosphoglycerate mutase 5 (PGAM5) functions as the convergent point for multiple necrosis pathways. This study aims to investigate whether inhibition of PGAM5 could reduce I/R-induced myocardial necroptosis and the underlying mechanisms. METHODS: The SD rat hearts (or H9c2 cells) were subjected to 1-h ischemia (or 10-h hypoxia) plus 3-h reperfusion (or 4-h reoxygenation) to establish the I/R (or H/R) injury model. The myocardial injury was assessed by the methods of biochemistry, H&E (hematoxylin and eosin), and PI/DAPI (propidium iodide/4',6-diamidino-2-phenylindole) staining, respectively. Drug interventions or gene knockdown was used to verify the role of PGAM5 in I/R (or H/R)-induced myocardial necroptosis and possible mechanisms. RESULTS: The I/R-treated heart showed the injuries (increase in infarct size and creatine kinase release), upregulation of PGAM5, dynamin-related protein 1 (Drp1), p-Drp1-S616, and necroptosis-relevant proteins (RIPK1/RIPK3, receptor-interacting protein kinase 1/3; MLKL, mixed lineage kinase domain-like); these phenomena were attenuated by inhibition of PGAM5 or RIPK1. In H9c2 cells, H/R treatment elevated the levels of PGAM5, RIPK1, RIPK3, MLKL, Drp1, and p-Drp1-S616 and induced mitochondrial dysfunctions (elevation in mitochondrial membrane potential and ROS level) and cellular necrosis (increase in LDH release and the ratio of PI+/DAPI+ cells); these effects were blocked by inhibition or knockdown of PGAM5. CONCLUSIONS: Inhibition of PGAM5 can reduce necroptosis in I/R-treated rat hearts through suppression of Drp1; there is a positive feedback between RIPK1 and PGAM5, and PGAM5 might serve as a novel therapeutic target for prevention of myocardial I/R injury.


Assuntos
DNA (Citosina-5-)-Metiltransferase 1/metabolismo , Inibidores Enzimáticos/farmacologia , Glicolatos/farmacologia , Proteínas Mitocondriais/antagonistas & inibidores , Infarto do Miocárdio/prevenção & controle , Traumatismo por Reperfusão Miocárdica/prevenção & controle , Miócitos Cardíacos/efeitos dos fármacos , Fosfoglicerato Mutase/antagonistas & inibidores , Fosfoproteínas Fosfatases/antagonistas & inibidores , Animais , Morte Celular/efeitos dos fármacos , Linhagem Celular , Modelos Animais de Doenças , Regulação para Baixo , Masculino , Proteínas Mitocondriais/genética , Proteínas Mitocondriais/metabolismo , Infarto do Miocárdio/enzimologia , Infarto do Miocárdio/patologia , Traumatismo por Reperfusão Miocárdica/enzimologia , Traumatismo por Reperfusão Miocárdica/patologia , Miócitos Cardíacos/enzimologia , Miócitos Cardíacos/patologia , Fosfoglicerato Mutase/genética , Fosfoglicerato Mutase/metabolismo , Fosfoproteínas Fosfatases/genética , Fosfoproteínas Fosfatases/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo , Ratos Sprague-Dawley , Transdução de Sinais/efeitos dos fármacos
7.
Heart Vessels ; 34(7): 1148-1157, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-30680494

RESUMO

A previous clinical study revealed elevation of chymase- and cathepsin G-dependent angiotensin II-forming activity (AIIFA) in the myocardium after acute myocardial infarction (AMI). This study examined the time course of chymase- and cathepsin G-dependent AIIFA in circulating mononuclear leukocytes (CML) after AMI. Consecutive patients with AMI were recruited. Chymase- and cathepsin G-dependent AIIFA in CML were assayed using a modified angiotensin I substrate with Nma/Dnp fluorescence quenching. The changes of CML AIIFA were monitored over time in the patients. Fifteen consecutive AMI patients admitted to our hospital were recruited. At 1 day after the admission, CML chymase- and cathepsin G-dependent AIIFA were 2.9- and 1.7-fold higher than at discharge, respectively. The ratio of chymase-dependent AIIFA to total AIIFA was significantly increased. AIIFA gradually decreased over time after the admission. The peak value of chymase- and cathepsin G-dependent AIIFA was significantly correlated with the maximum levels of aspartate aminotransferase (r = 0.53, 0.64), lactate dehydrogenase (r = 0.57, 0.62), and creatine kinase (r = 0.60, 0.65). This is the first evidence that chymase- and cathepsin G-dependent AIIFA is elevated in CML after AMI. Our data suggested that chymase-dependent AIIFA is increased in CML as well as in the myocardium after AMI, and that the level of chymase-dependent AIIFA might reflect the severity of infarction.


Assuntos
Angiotensina II/metabolismo , Catepsina G/metabolismo , Quimases/metabolismo , Infarto do Miocárdio/enzimologia , Miocárdio/patologia , Idoso , Estudos Transversais , Feminino , Humanos , Japão , Masculino , Pessoa de Meia-Idade , Infarto do Miocárdio/terapia , Miocárdio/enzimologia , Intervenção Coronária Percutânea
8.
J Cardiol ; 73(1): 81-88, 2019 01.
Artigo em Inglês | MEDLINE | ID: mdl-30487059

RESUMO

BACKGROUND: The hypothalamic paraventricular nucleus (PVN) is the center of the regulation of autonomic nervous system functions and cardiovascular activity. Phosphoinositide-3 kinase (PI3K)-AKT pathway in PVN contributes to mediate sympathetic nerve activity and is activated in spontaneously hypertensive rats. Overactivation of the sympathetic output was considered as an important mechanism of the arrhythmias. In the present study, we aimed to explore whether targeted regulation of sympathetic activity in PVN could reduce the peripheral sympathoexcitatory and attenuate the ventricular arrhythmias (VAs) in myocardial infarction (MI) rats via PI3K-AKT pathway. METHODS: A stainless steel gauge guide cannula was stereotaxically implanted into the PVN, and 7 days later, rats were randomly divided into the following 4 groups: group A, control+dimethyl sulfoxide (DMSO); group B, control+LY294002; group C, MI surgery+DMSO; and group D, MI surgery+LY294002. Studies were conducted seven days post-MI. Myocardial function, infarct size, inducible VAs by programmed electrical stimulation, renal sympathetic nerve activity (RSNA), and protein level of PI3K and AKT were measured. RESULTS: MI increased the protein ratios of p-PI3K-to-total-PI3K and p-AKT-to-total-AKT in PVN but can be reduced by LY294002 treatment. Inhibition of sympathetic nerve activity in PVN led to a reversion in plasma norepinephrine, RSNA and inducible VAs in MI rats. CONCLUSIONS: PI3K-AKT pathway in the PVN was a main mechanism in regulating sympathetic activity and arrhythmias in MI rats. Targeted inhibition of sympathetic activity in PVN may be a potential treatment for the VAs via PI3K-AKT pathway.


Assuntos
Arritmias Cardíacas/enzimologia , Infarto do Miocárdio/enzimologia , Núcleo Hipotalâmico Paraventricular/enzimologia , Fosfatidilinositol 3-Quinases/metabolismo , Sistema Nervoso Simpático/enzimologia , Animais , Arritmias Cardíacas/fisiopatologia , Arritmias Cardíacas/prevenção & controle , Cromonas/uso terapêutico , Inibidores Enzimáticos/uso terapêutico , Masculino , Morfolinas/uso terapêutico , Infarto do Miocárdio/tratamento farmacológico , Infarto do Miocárdio/fisiopatologia , Miocárdio/enzimologia , Norepinefrina/sangue , Ratos , Ratos Endogâmicos SHR , Transdução de Sinais/efeitos dos fármacos
9.
J Cardiovasc Pharmacol ; 73(2): 82-91, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30531435

RESUMO

AIMS: Inhibition of brain angiotensin III by central infusion of aminopeptidase A (APA) inhibitor firibastat (RB150) inhibits sympathetic hyperactivity and heart failure in rats after myocardial infarction (MI). This study evaluated effectiveness of systemic treatment with firibastat compared with AT1R blocker, losartan. METHODS AND RESULTS: MI was induced by ligation of left coronary artery in male Wistar rats. Rats were treated from 1 to 5 weeks after MI in protocol 1 with vehicle, or firibastat at 50 mg/kg/d subcutaneously (s.c.) or 150 mg/kg/d oral, once daily, and in protocol 2, with vehicle, firibastat 150 mg/kg or losartan 50 mg/kg oral twice daily. At 5 weeks, left ventricle function was evaluated by echocardiography and Millar catheter. After MI, rats developed moderate severe heart failure. Both s.c. and oral firibastat inhibited brain APA and attenuated left ventricle dysfunction. Oral firibastat and losartan similarly improved left ventricular end diastolic pressure. However, whereas firibastat improved dP/dtmax, losartan lowered dP/dtmax and left ventricular peak systolic pressure, and increased plasma creatinine by ~50%. On the other hand, losartan more effectively inhibited cardiac fibrosis. CONCLUSION: Inhibition of the brain renin-angiotensin system by oral APA inhibitor is at least as effective as oral AT1R blocker to inhibit cardiac dysfunction after MI but without hypotension or renal dysfunction.


Assuntos
Bloqueadores do Receptor Tipo 1 de Angiotensina II/administração & dosagem , Angiotensina III/metabolismo , Encéfalo/efeitos dos fármacos , Dissulfetos/administração & dosagem , Inibidores Enzimáticos/administração & dosagem , Glutamil Aminopeptidase/antagonistas & inibidores , Insuficiência Cardíaca/prevenção & controle , Losartan/administração & dosagem , Infarto do Miocárdio/tratamento farmacológico , Ácidos Sulfônicos/administração & dosagem , Administração Oral , Animais , Encéfalo/enzimologia , Encéfalo/fisiopatologia , Modelos Animais de Doenças , Fibrose , Glutamil Aminopeptidase/metabolismo , Insuficiência Cardíaca/enzimologia , Insuficiência Cardíaca/etiologia , Insuficiência Cardíaca/fisiopatologia , Injeções Subcutâneas , Masculino , Infarto do Miocárdio/complicações , Infarto do Miocárdio/enzimologia , Infarto do Miocárdio/fisiopatologia , Ratos Wistar , Transdução de Sinais , Função Ventricular Esquerda/efeitos dos fármacos , Pressão Ventricular/efeitos dos fármacos , Remodelação Ventricular/efeitos dos fármacos
10.
J Mol Cell Cardiol ; 127: 44-56, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30465799

RESUMO

BACKGROUND: Extracellular matrix metabolism and cardiac cell death participate centrally in myocardial infarction (MI). This study tested the roles of collagenolytic cathepsin K (CatK) in post-MI left ventricular remodeling. METHODS AND RESULTS: Patients with acute MI had higher plasma CatK levels (20.49 ±â€¯7.07 pmol/L, n = 26) than those in subjects with stable angina pectoris (8.34 ±â€¯1.66 pmol/L, n = 28, P = .01) or those without coronary heart disease (6.63 ±â€¯0.84 pmol/L, n = 93, P = .01). CatK protein expression increases in mouse hearts at 7 and 28 days post-MI. Immunofluorescent staining localized CatK expression in cardiomyocytes, endothelial cells, fibroblasts, macrophages, and CD4+ T cells in infarcted mouse hearts at 7 days post-MI. To probe the direct participation of CatK in MI, we produced experimental MI in CatK-deficient mice (Ctsk-/-) and their wild-type (Ctsk+/+) littermates. CatK-deficiency yielded worsened cardiac function at 7 and 28 days post-MI, compared to Ctsk+/+ littermates (fractional shortening percentage: 5.01 ±â€¯0.68 vs. 8.62 ±â€¯1.04, P < .01, 7 days post-MI; 4.32 ±â€¯0.52 vs. 7.60 ±â€¯0.82, P < .01, 28 days post-MI). At 7 days post-MI, hearts from Ctsk-/- mice contained less CatK-specific type-I collagen fragments (10.37 ±â€¯1.91 vs. 4.60 ±â€¯0.49 ng/mg tissue extract, P = .003) and more fibrosis (1.67 ±â€¯0.93 vs. 0.69 ±â€¯0.20 type-III collagen positive area percentage, P = .01; 14.25 ±â€¯4.12 vs. 6.59 ±â€¯0.79 α-smooth muscle actin-positive area percentage, P = .016; and 0.82 ±â€¯0.06 vs. 0.31 ±â€¯0.08 CD90-positive area percentage, P = .008) than those of Ctsk+/+ mice. Immunostaining demonstrated that CatK-deficiency yielded elevated cardiac cell death but reduced cardiac cell proliferation. In vitro studies supported a role of CatK in cardiomyocyte survival. CONCLUSION: Plasma CatK levels are increased in MI patients. Heart CatK expression is also elevated post-MI, but CatK-deficiency impairs post-MI cardiac function in mice by increasing myocardial fibrosis and cardiomyocyte death.


Assuntos
Catepsina K/deficiência , Testes de Função Cardíaca , Infarto do Miocárdio/enzimologia , Infarto do Miocárdio/fisiopatologia , Síndrome Coronariana Aguda/sangue , Síndrome Coronariana Aguda/enzimologia , Síndrome Coronariana Aguda/fisiopatologia , Idoso , Animais , Apoptose , Catepsina K/sangue , Proliferação de Células , Colágeno/metabolismo , Feminino , Fibrose , Ventrículos do Coração/metabolismo , Humanos , Inflamação/patologia , Masculino , Camundongos , Pessoa de Meia-Idade
11.
J Cardiovasc Pharmacol ; 73(2): 70-81, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30422891

RESUMO

Previous studies have shown that κ-opioid receptor activation possesses cardioprotection against myocardial ischemia and reperfusion (MI/R) injury. The current study was designed to investigate whether mitochondrial dysfunction after MI/R is regulated by the κ-opioid receptor and to further explore the underlying mechanisms involved. MI/R rat model was established in vivo, and a hypoxia and reoxygenation cardiomyocytes model was used in vitro. Mitochondrial morphology and function as well as myocardial apoptosis were determined. Our data indicated that treatment with U50,488H (a selective κ-opioid receptor agonist) not only reduced apoptosis but also significantly improved mitochondrial morphology and function. These effects were blocked by nor-binaltorphimine (nor-BNI, a selective κ-opioid receptor antagonist), Compound C (an AMPK inhibitor), and AR-A014418 (a GSK3ß inhibitor). Moreover, in cardiomyocytes, treatment with U50,488H significantly increased the expression in phosphorylation of AMPK and the phosphorylation of GSK3ß. Treatment of cardiomyocytes with AMPKα siRNA decreased the phosphorylation of AMPK and GSK3ß. Moreover, AMPK activation resulted in the phosphorylation of GSK3ß. Our findings suggested that U50,488H exerted cardioprotective effects by improving mitochondrial morphology and function against MI/R injury through activation of the κ-opioid receptor-mediated AMPK/GSK3ß pathway.


Assuntos
(trans)-Isômero de 3,4-dicloro-N-metil-N-(2-(1-pirrolidinil)-ciclo-hexil)-benzenoacetamida/farmacologia , Proteínas Quinases Ativadas por AMP/metabolismo , Apoptose/efeitos dos fármacos , Glicogênio Sintase Quinase 3 beta/metabolismo , Mitocôndrias Cardíacas/efeitos dos fármacos , Infarto do Miocárdio/prevenção & controle , Traumatismo por Reperfusão Miocárdica/prevenção & controle , Miócitos Cardíacos/efeitos dos fármacos , Receptores Opioides kappa/agonistas , Proteínas Quinases Ativadas por AMP/genética , Animais , Linhagem Celular , Modelos Animais de Doenças , Masculino , Mitocôndrias Cardíacas/enzimologia , Mitocôndrias Cardíacas/ultraestrutura , Infarto do Miocárdio/enzimologia , Infarto do Miocárdio/genética , Infarto do Miocárdio/patologia , Traumatismo por Reperfusão Miocárdica/enzimologia , Traumatismo por Reperfusão Miocárdica/genética , Traumatismo por Reperfusão Miocárdica/patologia , Miócitos Cardíacos/enzimologia , Miócitos Cardíacos/ultraestrutura , Fosforilação , Ratos Sprague-Dawley , Receptores Opioides kappa/metabolismo , Transdução de Sinais
12.
Clin Exp Hypertens ; 41(1): 62-69, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-29595329

RESUMO

BACKGROUND: Sacubitril (SAC), a neprilysin inhibitor prevent degradation of neprilysin and activate cGMP signaling pathways leading to rise in blood volume concurrent to blood pressure by means of vasoactive peptides, adrenomedullin, and bradykinin. OBJECTIVE: The aim of this study was to evaluate the anti-ischemic effects of SAC through inhibiting neprilysin in isoproterenol (ISO) induced myocardial infarction (MI) in Wistar albino rats. ISO (85 mg/kg) was injected subcutaneously at the end of 14 days pre-treatment with SAC and valsartan (VAL). RESULT: Biochemical investigation revealed that SAC along with VAL significantly prevented the antioxidant enzymes (SOD, Catalase, GR, GPx, GST, and GSH) degradation and malondialdehyde (MDA) induced by ISO intoxication in Wistar rats. Along with this, cardiac biomarkers (LDH, CK-MB, ALT, AST, and ALP) were also significantly ameliorated by SACand VAL in ISO-treated rats. Concurrently, decreased infarction area (IA)and marked reduction in myofibril damage by SACand VAL further supported its protective benefits in MI. CONCLUSION: Taken together, the results suggest that inhibition of enzyme neprilysin alleviated the ISO induces myocardial damage mediated by its strong antioxidant potential.


Assuntos
Aminobutiratos/farmacologia , Anti-Hipertensivos/farmacologia , Infarto do Miocárdio/enzimologia , Infarto do Miocárdio/prevenção & controle , Miocárdio/enzimologia , Neprilisina/antagonistas & inibidores , Tetrazóis/farmacologia , Valsartana/farmacologia , Alanina Transaminase/metabolismo , Fosfatase Alcalina/metabolismo , Aminobutiratos/uso terapêutico , Animais , Anti-Hipertensivos/uso terapêutico , Antioxidantes/farmacologia , Aspartato Aminotransferases/metabolismo , Catalase/metabolismo , Creatina Quinase Forma MB/metabolismo , Glutationa Peroxidase/metabolismo , Glutationa Redutase/metabolismo , Glutationa Transferase/metabolismo , Isoproterenol , Lactato Desidrogenases/metabolismo , Masculino , Malondialdeído/metabolismo , Infarto do Miocárdio/induzido quimicamente , Estresse Oxidativo/efeitos dos fármacos , Ratos , Ratos Wistar , Superóxido Dismutase/metabolismo , Tetrazóis/uso terapêutico , Valsartana/uso terapêutico
13.
Circulation ; 139(4): 518-532, 2019 01 22.
Artigo em Inglês | MEDLINE | ID: mdl-29997116

RESUMO

BACKGROUND: Despite its functional importance in various fundamental bioprocesses, studies of N6-methyladenosine (m6A) in the heart are lacking. Here, we show that the FTO (fat mass and obesity-associated protein), an m6A demethylase, plays a critical role in cardiac contractile function during homeostasis, remodeling, and regeneration. METHODS: We used clinical human samples, preclinical pig and mouse models, and primary cardiomyocyte cell cultures to study the functional role of m6A and FTO in the heart and in cardiomyocytes. We modulated expression of FTO by using adeno-associated virus serotype 9 (in vivo), adenovirus (both in vivo and in vitro), and small interfering RNAs (in vitro) to study its function in regulating cardiomyocyte m6A, calcium dynamics and contractility, and cardiac function postischemia. We performed methylated (m6A) RNA immunoprecipitation sequencing to map transcriptome-wide m6A, and methylated (m6A) RNA immunoprecipitation quantitative polymerase chain reaction assays to map and validate m6A in individual transcripts, in healthy and failing hearts, and in myocytes. RESULTS: We discovered that FTO has decreased expression in failing mammalian hearts and hypoxic cardiomyocytes, thereby increasing m6A in RNA and decreasing cardiomyocyte contractile function. Improving expression of FTO in failing mouse hearts attenuated the ischemia-induced increase in m6A and decrease in cardiac contractile function. This is performed by the demethylation activity of FTO, which selectively demethylates cardiac contractile transcripts, thus preventing their degradation and improving their protein expression under ischemia. In addition, we demonstrate that FTO overexpression in mouse models of myocardial infarction decreased fibrosis and enhanced angiogenesis. CONCLUSIONS: Collectively, our study demonstrates the functional importance of the FTO-dependent cardiac m6A methylome in cardiac contraction during heart failure and provides a novel mechanistic insight into the therapeutic mechanisms of FTO.


Assuntos
Adenosina/análogos & derivados , Insuficiência Cardíaca/enzimologia , Infarto do Miocárdio/enzimologia , Miócitos Cardíacos/enzimologia , Regeneração , Função Ventricular Esquerda , Remodelação Ventricular , Adenosina/metabolismo , Adulto , Idoso , Dioxigenase FTO Dependente de alfa-Cetoglutarato/genética , Dioxigenase FTO Dependente de alfa-Cetoglutarato/metabolismo , Animais , Sinalização do Cálcio , Estudos de Casos e Controles , Linhagem Celular , Proliferação de Células , Desmetilação , Modelos Animais de Doenças , Feminino , Insuficiência Cardíaca/genética , Insuficiência Cardíaca/patologia , Insuficiência Cardíaca/fisiopatologia , Humanos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Pessoa de Meia-Idade , Infarto do Miocárdio/genética , Infarto do Miocárdio/patologia , Infarto do Miocárdio/fisiopatologia , Miócitos Cardíacos/patologia , Processamento Pós-Transcricional do RNA , Estabilidade de RNA , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos Sprague-Dawley , Sus scrofa
14.
J Cardiovasc Pharmacol Ther ; 24(1): 78-89, 2019 01.
Artigo em Inglês | MEDLINE | ID: mdl-30033751

RESUMO

Cardiac steroids (CSs), such as ouabain and digoxin, increase the force of contraction of heart muscle and are used for the treatment of congestive heart failure (CHF). However, their small therapeutic window limits their use. It is well established that Na+, K+-ATPase inhibition mediates CS-induced increase in heart contractility. Recently, the involvement of intracellular signal transduction was implicated in this effect. The aim of the present study was to test the hypothesis that combined treatment with ouabain and Akt inhibitor (MK-2206) augments ouabain-induced inotropy in mammalian models. We demonstrate that the combined treatment led to an ouabain-induced increase in contractility at concentrations at which ouabain alone was ineffective. This was shown in 3 experimental systems: neonatal primary rat cardiomyocytes, a Langendorff preparation, and an in vivo myocardial infarction induced by left anterior descending coronary artery (LAD) ligation. Furthermore, cell viability experiments revealed that this treatment protected primary cardiomyocytes from MK-2206 toxicity and in vivo reduced the size of scar tissue 10 days post-LAD ligation. We propose that Akt activity imposes a constant inhibitory force on muscle contraction, which is attenuated by low concentrations of MK-2206, resulting in potentiation of the ouabain effect. This demonstration of the increase in the CS effect advocates the development of the combined treatment in CHF.


Assuntos
Cardiotônicos/farmacologia , Compostos Heterocíclicos com 3 Anéis/farmacologia , Contração Miocárdica/efeitos dos fármacos , Infarto do Miocárdio/tratamento farmacológico , Miócitos Cardíacos/efeitos dos fármacos , Ouabaína/farmacologia , Inibidores de Proteínas Quinases/farmacologia , Proteínas Proto-Oncogênicas c-akt/antagonistas & inibidores , Função Ventricular Esquerda/efeitos dos fármacos , Animais , Células Cultivadas , Modelos Animais de Doenças , Sinergismo Farmacológico , Quimioterapia Combinada , Preparação de Coração Isolado , Masculino , Infarto do Miocárdio/enzimologia , Infarto do Miocárdio/patologia , Infarto do Miocárdio/fisiopatologia , Miócitos Cardíacos/enzimologia , Miócitos Cardíacos/patologia , Proteínas Proto-Oncogênicas c-akt/metabolismo , Ratos Wistar , Transdução de Sinais
15.
Eur Rev Med Pharmacol Sci ; 22(24): 8990-8998, 2018 12.
Artigo em Inglês | MEDLINE | ID: mdl-30575944

RESUMO

OBJECTIVE: The aim of the study was to explore the pharmacological role of ulinastatin in rats with myocardium infarction (MI) and its underlying mechanism. MATERIALS AND METHODS: Rats were randomly assigned into sham group, MI group, and ulinastatin group, with 8 rats in each group. MI model in rats was constructed and specific drug administrations were performed in each group. Electrocardiogram and hemodynamics were detected before and after animal procedures. Myocardium function in rats was accessed by determining serum levels of creatine kinase-MB (CK-MB) and lactate dehydrogenase (LDH). Hematoxylin and eosin (HE) staining was performed to detect the infarct area in rat myocardium. Terminal Deoxynucleotidyl Transferase dUTP Nick-end Labeling (TUNEL) assay was conducted to calculate the ratio of apoptotic cells in rat myocardium. Subsequently, relative oxidative stress indicators in rats were accessed, including total antioxidant capacity (T-AOC), catalase (CAT), glutathione (GSH), Superoxide Dismutase (SOD), malondialdehyde (MDA), and reactive oxygen species (ROS). Western blot was conducted to detect protein levels of nuclear factor E2-related factor 2 (Nrf2), Nuclear Factor κB (NF-Κb), heme oxygenase-1 (HO-1), and NAD(P)H:quinone oxidoreductase 1 (NQO1) in rat myocardium. RESULTS: No significant differences in heart rate, the voltage of the QRS wave and Q-T interval were observed among rats in sham group, MI group, and ulinastatin group prior to the animal procedures. However, at the end of the animal procedures, rats in ulinastatin group showed higher heart rate and voltage of QRS wave, as well as shorter Q-T interval than those in MI group. Rats in ulinastatin group presented lower serum levels of CK-MB and LDH compared with those in MI group, whereas they did not return to baseline. Rats in ulinastatin group showed higher levels of LVSP, dP/dtmax, LVEDP, and -dP/dtmax than those in MI group. Larger infarct area was observed in MI group compared with that of sham group, whereas ulinastatin treatment remarkably reduced the infarct area. HE staining showed remarkable pathological lesions in MI rats, whereas ulinastatin group showed milder lesions in rat myocardium. TUNEL assay showed fewer TUNEL-positive cells in ulinastatin group than those of MI group. Levels of T-AOC, CAT, GSH, and SOD were remarkably higher in myocardium homogenate of MI group than sham group, whereas ulinastatin treatment significantly decreased these levels. Ulinastatin group showed less ROS accumulation and decreased MDA level in rats than those of MI group. Ulinastatin treatment upregulated Nrf2, HO-1, and NQO1, whereas downregulated NF-κB expression. CONCLUSIONS: Ulinastatin protects MI rats by inhibiting inflammatory response through activating Nrf2/NOS pathway.


Assuntos
Anti-Inflamatórios/farmacologia , Antioxidantes/farmacologia , Glicoproteínas/farmacologia , Hemodinâmica/efeitos dos fármacos , Infarto do Miocárdio/tratamento farmacológico , Miócitos Cardíacos/efeitos dos fármacos , Fator 2 Relacionado a NF-E2/metabolismo , Óxido Nítrico Sintase/metabolismo , Função Ventricular Esquerda/efeitos dos fármacos , Animais , Apoptose/efeitos dos fármacos , Modelos Animais de Doenças , Mediadores da Inflamação/metabolismo , Masculino , Infarto do Miocárdio/enzimologia , Infarto do Miocárdio/patologia , Infarto do Miocárdio/fisiopatologia , Miócitos Cardíacos/enzimologia , Miócitos Cardíacos/patologia , Estresse Oxidativo/efeitos dos fármacos , Ratos Sprague-Dawley , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais
16.
BMC Cardiovasc Disord ; 18(1): 196, 2018 10 20.
Artigo em Inglês | MEDLINE | ID: mdl-30342492

RESUMO

BACKGROUND: Receptor tyrosine kinases (RTK) are potential targets for the treatment of ischemic heart disease. The human RTK family consists of 55 members, most of which have not yet been characterized for expression or activity in the ischemic heart. METHODS: RTK gene expression was analyzed from human heart samples representing healthy tissue, acute myocardial infarction or ischemic cardiomyopathy. As an experimental model, pig heart with ischemia-reperfusion injury, caused by cardiopulmonary bypass, was used, from which phosphorylation status of RTKs was assessed with a phospho-RTK array. Expression and function of one RTK, ROR1, was further validated in pig tissue samples, and in HL-1 cardiomyocytes and H9c2 cardiomyoblasts, exposed to hypoxia and reoxygenation. ROR1 protein level was analyzed by Western blotting. Cell viability after ROR1 siRNA knockdown or activation with Wnt-5a ligand was assessed by MTT assays. RESULTS: In addition to previously characterized RTKs, a group of novel active and regulated RTKs was detected in the ischemic heart. ROR1 was the most significantly upregulated RTK in human ischemic cardiomyopathy. However, ROR1 phosphorylation was suppressed in the pig model of ischemia-reperfusion and ROR1 phosphorylation and expression were down-regulated in HL-1 cardiomyocytes subjected to short-term hypoxia in vitro. ROR1 expression in the pig heart was confirmed on protein and mRNA level. Functionally, ROR1 activity was associated with reduced viability of HL-1 cardiomyocytes in both normoxia and during hypoxia-reoxygenation. CONCLUSIONS: Several novel RTKs were found to be regulated in expression or activity in ischemic heart. ROR1 was one of the most significantly regulated RTKs. The in vitro findings suggest a role for ROR1 as a potential target for the treatment of ischemic heart injury.


Assuntos
Cardiomiopatias/enzimologia , Infarto do Miocárdio/enzimologia , Traumatismo por Reperfusão Miocárdica/enzimologia , Miócitos Cardíacos/enzimologia , Receptores Órfãos Semelhantes a Receptor Tirosina Quinase/metabolismo , Animais , Cardiomiopatias/tratamento farmacológico , Cardiomiopatias/genética , Cardiomiopatias/patologia , Estudos de Casos e Controles , Linhagem Celular , Sobrevivência Celular , Modelos Animais de Doenças , Regulação Enzimológica da Expressão Gênica , Humanos , Terapia de Alvo Molecular , Infarto do Miocárdio/tratamento farmacológico , Infarto do Miocárdio/genética , Infarto do Miocárdio/patologia , Traumatismo por Reperfusão Miocárdica/tratamento farmacológico , Traumatismo por Reperfusão Miocárdica/genética , Traumatismo por Reperfusão Miocárdica/patologia , Miócitos Cardíacos/efeitos dos fármacos , Miócitos Cardíacos/patologia , Fosforilação , Inibidores de Proteínas Quinases/farmacologia , Receptores Órfãos Semelhantes a Receptor Tirosina Quinase/antagonistas & inibidores , Receptores Órfãos Semelhantes a Receptor Tirosina Quinase/genética , Transdução de Sinais , Sus scrofa
17.
Eur Rev Med Pharmacol Sci ; 22(19): 6475-6484, 2018 10.
Artigo em Inglês | MEDLINE | ID: mdl-30338817

RESUMO

OBJECTIVE: The myocardial ischemia/reperfusion (I/R) injury is a significant challenge, and the clinical significance of remote ischemic postconditioning (RIPostC) in cardioprotection has been confirmed. However, the molecular mechanism remains unclear. We aimed to explore the regulatory mechanism of RIPostC in myocardial I/R. MATERIALS AND METHODS: A mouse model of myocardial I/R injury and cell model of oxygen-glucose deprivation (OGD)/re-oxygenation (OGD/R) injury were constructed. Infarct size was measured by Evans blue dye staining and TTC staining. mRNA and protein expression levels of aldehyde dehydrogenase 2 (ALDH2) were determined by RT-qPCR and Western blot analysis, respectively. Cell viability, p53 expression, apoptotic cells, expression of proteins related to apoptosis, and reactive oxygen species (ROS) generation were evaluated by CCK-8 assay, Western blot analysis, flow cytometry assay, Western blot analysis, and DCFH-DA staining, respectively. ALDH2 in H9c2 cells was knocked down, and its effects on cells treated with OGD/R and RIPostC were tested. How RIPostC affected ALDH2 expression was finally studied. RESULTS: RIPostC reduced infarct size in mice and attenuated OGD/R-induced H9c2 cell injury. Myocardial I/R-induced down-regulation of ALDH2 was abrogated by RIPostC. Moreover, the effects of RIPostC on OGD/R-treated H9c2 cells were significantly reversed by ALDH2 silence. Finally, we found RIPostC-induced up-regulation of ALDH2 in OGD/R-treated cells could be bated by activation of PI3K and/or mTOR. CONCLUSIONS: RIPostC exerted cardioprotective role against myocardial I/R both in vivo and in vitro. Up-regulation of ALDH2 might be a reason for the cardioprotection, and RIPostC might regulate ALDH2 expression via the PI3K/mTOR pathway.


Assuntos
Aldeído-Desidrogenase Mitocondrial/biossíntese , Apoptose , Artéria Femoral/cirurgia , Pós-Condicionamento Isquêmico/métodos , Infarto do Miocárdio/prevenção & controle , Traumatismo por Reperfusão Miocárdica/prevenção & controle , Miócitos Cardíacos/enzimologia , Aldeído-Desidrogenase Mitocondrial/genética , Animais , Proteínas Reguladoras de Apoptose/metabolismo , Hipóxia Celular , Linhagem Celular , Modelos Animais de Doenças , Indução Enzimática , Artéria Femoral/fisiopatologia , Glucose/deficiência , Ligadura , Masculino , Camundongos Endogâmicos C57BL , Infarto do Miocárdio/enzimologia , Infarto do Miocárdio/patologia , Traumatismo por Reperfusão Miocárdica/enzimologia , Traumatismo por Reperfusão Miocárdica/patologia , Miócitos Cardíacos/patologia , Fosfatidilinositol 3-Quinase/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Fluxo Sanguíneo Regional , Transdução de Sinais , Serina-Treonina Quinases TOR/metabolismo , Proteína Supressora de Tumor p53/metabolismo
18.
Discov Med ; 26(141): 7-20, 2018 08.
Artigo em Inglês | MEDLINE | ID: mdl-30265851

RESUMO

We aimed to study urotensin II (UII) in acute myocardial infarction (AMI) patients before and after percutaneous coronary intervention (PCI) and to investigate its interaction with angiotensin II (ATII) in post-AMI myocardial remodeling. In this clinical study, 63 AMI patients and 66 controls were included. Levels of UII, ATII, and NT-B-type natriuretic peptide (NT-BNP) were determined. Participants were followed up for 2 years to observe clinical outcomes. In cell biology experiments, ATII and UII were added into cardiomyocytes and fibroblasts individually or in combination. Effects of ATII and UII on cardiac hypertrophy and fibrosis were observed. UII levels increased significantly after AMI (P < 0.001), especially after PCI (P = 0.0009), and was an independent predictor of death (OR 0.007, P = 0.005). UII correlated negatively with hypertension risk stratifications (P = 0.047) and the number of coronary artery lesions (P = 0.029), whereas it correlated positively with left ventricular ejection fraction (P = 0.0395). Myocardial ERK phosphorylations (P = 0.0041) were promoted and the expressions of cardiac hypertrophy-related genes (BNP, ANP) were upregulated in cells treated with ATII and UII (P < 0.005). The ATII+UII group also had significant increases of matrix metalloproteinase-2 (MMP-2), metalloproteinase-9 (MMP9), and fibronectin expressions, as well as collagen type I, III, MMP2, and MMP9 gene transcriptions (all P < 0.05). The UII level rose in AMI and was a predictor of death, but at the right concentration it may also have a cardioprotective role. Our findings suggest that UII and ATII have synergistic effects on cardiomyocyte remodeling. UII may play a role in the myocardial healing process post-AMI.


Assuntos
Infarto do Miocárdio/sangue , Urotensinas/sangue , Angiotensina II/sangue , Animais , Animais Recém-Nascidos , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Fibroblastos/metabolismo , Fibroblastos/patologia , Humanos , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/metabolismo , Infarto do Miocárdio/enzimologia , Infarto do Miocárdio/fisiopatologia , Miocárdio/patologia , Peptídeo Natriurético Encefálico/sangue , Fragmentos de Peptídeos/sangue , Fosforilação , Valor Preditivo dos Testes , Ratos Wistar , Sensibilidade e Especificidade , Volume Sistólico , Regulação para Cima
19.
Sci Rep ; 8(1): 11820, 2018 08 07.
Artigo em Inglês | MEDLINE | ID: mdl-30087386

RESUMO

Cardiac surgery with cardiopulmonary bypass (CPB) triggers myocardial ischemia/reperfusion injury contributing to organ dysfunction. Preclinical studies revealed that dipeptidyl peptidase (DPP4) inhibition is protective during myocardial infarction. Here, we assessed for the first time the relation of peri-operative DPP4-activity in serum of 46 patients undergoing cardiac surgery with patients' post-operative organ dysfunction during intensive care unit (ICU) stay. Whereas a prior myocardial infarction significantly reduced pre-operative DDP4-activity, patients with preserved left ventricular function showed an intra-operative decrease of DPP4-activity. The latter correlated with aortic cross clamping time, indicative for the duration of surgery-induced myocardial ischemia. As underlying mechanism, mass-spectrometry revealed increased DPP4 oxidation by cardiac surgery, with DPP4 oxidation reducing DPP4-activity in vitro. Further, post-operative DPP4-activity was negatively correlated with the extent of post-operative organ injury as measured by SAPS II and SOFA scoring, circulating levels of creatinine and lactate, as well as patients' stay on the ICU. In conclusion, cardiac surgery reduces DPP4-activity through oxidation, with low post-operative DPP4-activity being associated with organ dysfunction and worse outcome of patients during the post-operative ICU stay. This likely reflects the severity of myocardial ischemia/reperfusion injury and may suggest potential beneficial effects of anti-oxidative treatments during cardiac surgery.


Assuntos
Procedimentos Cirúrgicos Cardíacos/métodos , Dipeptidil Peptidase 4/metabolismo , Unidades de Terapia Intensiva , Infarto do Miocárdio/fisiopatologia , Traumatismo por Reperfusão Miocárdica/fisiopatologia , Idoso , Procedimentos Cirúrgicos Cardíacos/efeitos adversos , Dipeptidil Peptidase 4/sangue , Feminino , Humanos , Tempo de Internação , Masculino , Pessoa de Meia-Idade , Infarto do Miocárdio/sangue , Infarto do Miocárdio/enzimologia , Traumatismo por Reperfusão Miocárdica/enzimologia , Traumatismo por Reperfusão Miocárdica/etiologia , Oxirredução , Período Pós-Operatório , Estudos Prospectivos
20.
Am J Physiol Heart Circ Physiol ; 315(5): H1148-H1158, 2018 11 01.
Artigo em Inglês | MEDLINE | ID: mdl-30074840

RESUMO

Epoxyeicosatrienoic acids (EETs) decrease cardiac ischemia-reperfusion injury; however, the mechanism of their protective effect remains elusive. Here, we investigated the cardioprotective action of a novel EET analog, EET-B, in reperfusion and the role of hypoxia-inducible factor (HIF)-1α in such action of EET-B. Adult male rats were subjected to 30 min of left coronary artery occlusion followed by 2 h of reperfusion. Administration of 14,15-EET (2.5 mg/kg) or EET-B (2.5 mg/kg) 5 min before reperfusion reduced infarct size expressed as a percentage of the area at risk from 64.3 ± 1.3% in control to 42.6 ± 1.9% and 46.0 ± 1.6%, respectively, and their coadministration did not provide any stronger effect. The 14,15-EET antagonist 14,15-epoxyeicosa-5( Z)-enoic acid (2.5 mg/kg) inhibited the infarct size-limiting effect of EET-B (62.5 ± 1.1%). Similarly, the HIF-1α inhibitors 2-methoxyestradiol (2.5 mg/kg) and acriflavine (2 mg/kg) completely abolished the cardioprotective effect of EET-B. In a separate set of experiments, the immunoreactivity of HIF-1α and its degrading enzyme prolyl hydroxylase domain protein 3 (PHD3) were analyzed in the ischemic areas and nonischemic septa. At the end of ischemia, the HIF-1α immunogenic signal markedly increased in the ischemic area compared with the septum (10.31 ± 0.78% vs. 0.34 ± 0.08%). After 20 min and 2 h of reperfusion, HIF-1α immunoreactivity decreased to 2.40 ± 0.48% and 1.85 ± 0.43%, respectively, in the controls. EET-B blunted the decrease of HIF-1α immunoreactivity (7.80 ± 0.69% and 6.44 ± 1.37%, respectively) and significantly reduced PHD3 immunogenic signal in ischemic tissue after reperfusion. In conclusion, EET-B provides an infarct size-limiting effect at reperfusion that is mediated by HIF-1α and downregulation of its degrading enzyme PHD3. NEW & NOTEWORTHY The present study shows that EET-B is an effective agonistic 14,15-epoxyeicosatrienoic acid analog, and its administration before reperfusion markedly reduced myocardial infarction in rats. Most importantly, we demonstrate that increased hypoxia-inducible factor-1α levels play a role in cardioprotection mediated by EET-B in reperfusion likely by mechanisms including downregulation of the hypoxia-inducible factor -1α-degrading enzyme prolyl hydroxylase domain protein 3.


Assuntos
Ácido 8,11,14-Eicosatrienoico/farmacologia , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Prolina Dioxigenases do Fator Induzível por Hipóxia/metabolismo , Infarto do Miocárdio/prevenção & controle , Traumatismo por Reperfusão Miocárdica/prevenção & controle , Miocárdio/enzimologia , Função Ventricular Esquerda/efeitos dos fármacos , Remodelação Ventricular/efeitos dos fármacos , Ácido 8,11,14-Eicosatrienoico/análogos & derivados , Ácido 8,11,14-Eicosatrienoico/uso terapêutico , Animais , Modelos Animais de Doenças , Regulação para Baixo , Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , Prolina Dioxigenases do Fator Induzível por Hipóxia/genética , Masculino , Infarto do Miocárdio/enzimologia , Infarto do Miocárdio/patologia , Infarto do Miocárdio/fisiopatologia , Traumatismo por Reperfusão Miocárdica/enzimologia , Traumatismo por Reperfusão Miocárdica/patologia , Traumatismo por Reperfusão Miocárdica/fisiopatologia , Miocárdio/patologia , Proteólise , Ratos Sprague-Dawley , Transdução de Sinais/efeitos dos fármacos
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