RESUMEN
BACKGROUND: Aconiti Lateralis Radis Praeparata (FZ, Fuzi) and Angelicae Sinensis Radix (DG, Danggui) are widely used in traditional Chinese medicine for the treatment of coronary heart disease. However, the protective effects of their combination on cardiomyocytes remain unexplored. This study aimed to investigate whether the combined application of these 2 herbs confers protective benefits on H9c2 cardiomyocytes subjected to hypoxia/reoxygenation (H/R) injury. METHODS: A H/R model (6h hypoxia followed by 6h reoxygenation) was established using H9c2 cardiomyocytes. The cells were randomly assigned to the following groups: control, model, FZ, DG, FZDG 3 mg/mL, FZDG 9 mg/mL, FZDG 27 mg/mL, FZDG 27 mg/mLâ +â LY, and LYâ +â model. Cell viability was assessed using the cell counting kit-8 assay, and apoptosis was measured by Annexin V-FITC/PI staining. The levels of lactate dehydrogenase, malondialdehyde, and superoxide dismutase were determined using colorimetric methods. The distribution and expression of apoptosis-related proteins were evaluated via immunohistochemistry. Protein and mRNA expression levels of key components in the phosphatidylinositol 3-kinase/protein kinase B/glycogen synthase kinase-3ß (PI3K/Akt/GSK-3ß) and autophagy pathways were analyzed by Western blot and reverse transcription-polymerase chain reaction, respectively. RESULTS: This study demonstrated that the FZDG 27 mg/mL treatment significantly enhanced cell viability and reduced the overall rate of apoptosis. This group also ameliorated oxidative stress injury in H/R cardiomyocytes, as evidenced by decreased levels of lactate dehydrogenase and malondialdehyde, along with an increase in superoxide dismutase activity. Moreover, the expression and distribution of key proteins were modulated, with elevated phosphorylation levels of Akt and GSK-3ß, as well as altered expression of B-cell lymphoma-2 and B-cell lymphoma-2-associated X protein. Western blot and reverse transcription-polymerase chain reaction demonstrated that the FZDG 27 mg/mL group most potently upregulated the PI3K/Akt/GSK-3ß and autophagy pathways among all groups, an effect blocked by LY294002. CONCLUSION: This study demonstrated that the combination of Aconiti Lateralis Radix Praeparata and Angelicae Sinensis significantly protected H/R cardiomyocytes in vitro. This protective effect was achieved through enhancing cell viability, reducing oxidative stress-induced injury, and modulating the expression and distribution of key proteins and genes. The underlying mechanisms involved the activation of the PI3K/Akt/GSK-3ß signaling pathway and the autophagy pathway, as confirmed by the inhibitory effect of LY294002.
Asunto(s)
Aconitum , Angelica sinensis , Medicamentos Herbarios Chinos , Daño por Reperfusión Miocárdica , Miocitos Cardíacos , Miocitos Cardíacos/efectos de los fármacos , Miocitos Cardíacos/metabolismo , Autofagia/efectos de los fármacos , Glucógeno Sintasa Quinasa 3 beta/metabolismo , Medicamentos Herbarios Chinos/farmacología , Proteínas Proto-Oncogénicas c-akt/metabolismo , Animales , Ratas , Fosfatidilinositol 3-Quinasas/metabolismo , Apoptosis/efectos de los fármacos , Transducción de Señal/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Línea Celular , Daño por Reperfusión Miocárdica/prevención & control , Daño por Reperfusión Miocárdica/tratamiento farmacológicoRESUMEN
Diabetes mellitus increases the heart's vulnerability to ischemia/reperfusion (I/R) injury and limits the efficacy of traditional cardioprotective interventions. This study evaluated the cardioprotective potential of 5'-N-ethylcarboxamido adenosine (NECA), a non-selective adenosine receptor agonist, in a diabetic rat model of myocardial I/R injury and explored the underlying molecular mechanisms. Type 2 diabetes was induced in Sprague-Dawley rats by high-fat diet feeding and streptozotocin injection. Myocardial I/R injury was established via 30-min coronary ligation followed by 2-h reperfusion. Rats were randomized into eleven treatment groups, including NECA, receptor modulators, and kinase regulators. Cardiac function and myocardial injury were assessed by echocardiography, TTC/Evans blue staining, histopathology, and plasma cardiac troponin I (cTnI) levels. NECA significantly improved left ventricular systolic function, reduced infarct size, and decreased cTnI release. These protective effects were associated with increased Bcl-2 expression and suppressed levels of cleaved caspase-3 and miR-15a. Pharmacological blockade of the A2A receptor with ZM241385 abolished NECA's benefits, while the A2AR agonist CGS21680 and PKCα activator PMA reproduced them. Electrophoretic mobility shift assays confirmed a direct interaction between PKCα and pri-miR-15a, suggesting transcriptional regulation. Collectively, these findings demonstrate that NECA attenuates myocardial I/R injury in diabetic rats through activation of the A2AR-PKCα signaling axis and inhibition of miR-15a, offering a potential therapeutic strategy for diabetic patients at risk of ischemic cardiac events.
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Agonistas del Receptor de Adenosina A2 , Cardiotónicos , Diabetes Mellitus Experimental , Diabetes Mellitus Tipo 2 , Poscondicionamiento Isquémico , Daño por Reperfusión Miocárdica , Proteína Quinasa C-alfa , Receptor de Adenosina A2A , Animales , Ratas Sprague-Dawley , Daño por Reperfusión Miocárdica/tratamiento farmacológico , Daño por Reperfusión Miocárdica/metabolismo , Daño por Reperfusión Miocárdica/patología , Daño por Reperfusión Miocárdica/prevención & control , Masculino , Diabetes Mellitus Experimental/tratamiento farmacológico , Diabetes Mellitus Experimental/complicaciones , Transducción de Señal/efectos de los fármacos , Ratas , Proteína Quinasa C-alfa/metabolismo , Receptor de Adenosina A2A/metabolismo , Fenetilaminas/farmacología , Fenetilaminas/uso terapéutico , Diabetes Mellitus Tipo 2/tratamiento farmacológico , Diabetes Mellitus Tipo 2/complicaciones , Adenosina/análogos & derivados , Adenosina/farmacología , Adenosina/uso terapéutico , Humanos , Miocardio/patología , Miocardio/metabolismo , MicroARNs/metabolismo , MicroARNs/genética , Cardiotónicos/uso terapéutico , Cardiotónicos/farmacología , Agonistas del Receptor de Adenosina A2/uso terapéutico , Agonistas del Receptor de Adenosina A2/farmacología , Troponina I/sangreRESUMEN
Myocardial ischemia/reperfusion (I/R) injury remains a major clinical challenge, with ferroptosis emerging as a critical contributor to cardiomyocyte death. Notably, ring-finger protein 5 (RNF5), which functions as an E3 ubiquitin ligase that is recognized for its crucial involvement in the modulation of endoplasmic reticulum stress, unfolded protein responses, and inflammatory processes, has not been systematically investigated in the process of the onset and evolutionary trajectory of myocardial ischemia/reperfusion (MI/R) injury.Male 7-8 week-oldC57BL/6 mice were used to establish a myocardial injury model of ischemia/reperfusion injury. Adeno-associated virus serotype 9 (AAV9) was used to regulate the expression of RNF5 in vivo and in vitro. Western blot analysis and immunohistochemistry (IHC) were employed to assess the expression levels of RNF5. A TUNEL assay was used to detect the apoptosis of myocardial cells. Immunoprecipitation-mass spectrometry (IP-MS) and coimmunoprecipitation (co-IP) were used to identify the proteins that interact with RNF5 to further explore the underlying mechanism involved.RNF5 was significantly downregulated in the myocardial tissues of MI/R mice. RNF5 overexpression alleviated myocardial tissue damage, reduced ferroptosis levels, and mitigated oxidative stress injury in MI/R model mice. Moreover, RNF5 overexpression markedly suppressed ROS levels, oxidative stress damage, and apoptosis in H9C2 cells subjected to H/R. Mechanistically, we found that RNF5 interacted with Acyl-coA synthetase long-chain family member 4(ACSL4) via its transmembrane region and mediated the degradation of ACSL4, thereby inhibiting the ferroptosis of cardiomyocytes.Our findings reveal a previously unrecognized ubiquitin-dependent mechanism controlling ferroptosis in the heart and position RNF5 as a promising therapeutic target for ischemic cardiomyopathy.
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Coenzima A Ligasas , Proteínas de Unión al ADN , Ferroptosis , Daño por Reperfusión Miocárdica , Ubiquitina-Proteína Ligasas , Ubiquitinación , Animales , Ferroptosis/fisiología , Ferroptosis/efectos de los fármacos , Daño por Reperfusión Miocárdica/metabolismo , Daño por Reperfusión Miocárdica/prevención & control , Daño por Reperfusión Miocárdica/patología , Masculino , Ratones , Ratones Endogámicos C57BL , Coenzima A Ligasas/metabolismo , Coenzima A Ligasas/genética , Ubiquitinación/fisiología , Ubiquitina-Proteína Ligasas/metabolismo , Ubiquitina-Proteína Ligasas/genética , Ligasa de Acido Graso-CoA de Cadena Larga , Miocitos Cardíacos/metabolismo , Miocitos Cardíacos/patología , Humanos , Proteínas de Unión al ADN/metabolismo , Proteínas de Unión al ADN/genéticaRESUMEN
Ischemia/reperfusion (I/R) injury causes myocardial damage, and Ginkgolide C (GC), a component of Ginkgo biloba, shows potential for cardioprotection. However, its effects on I/R-induced cardiac dysfunction and mechanisms are unclear. This study evaluates GC preconditioning in rats, focusing on its impact on cardiac function, myocardial injury, and the PI3K/Akt pathway. GC preconditioning was assessed using an isolated rat heart model of I/R injury. Cardiac function (LVDP, LVEDP, +dp/dtmax, -dp/dtmax), infarct size, histopathological changes, injury, and oxidative stress markers were measured. Rat cardiomyocytes were treated with GC to assess viability, contraction, and apoptosis. Molecular docking and protein analysis explored GC's interaction with the Akt pathway. GC preconditioning significantly improved cardiac function, with a marked enhancement in LVDP, +dp/dtmax, and -dp/dtmax in the GC50 group compared to the I/R group (P<0.01). GC treatment also reduced myocardial infarct size (14.8 %±2.4 % vs. 25.5 %±1.9 %, P<0.01), decreased LDH release, and alleviated histopathological damage, including myocyte necrosis and inflammatory infiltration. Furthermore, GC significantly improved cardiomyocyte viability and contraction amplitude, particularly at 50 ?M. At the molecular level, GC activated the Akt pathway, as evidenced by increased p-Akt expression. Co-treatment with the PI3K inhibitor LY294002 reversed the protective effects of GC, confirming the involvement of the Akt pathway. Additionally, GC preconditioning reduced oxidative stress, as indicated by enhanced SOD activity and decreased levels of myocardial injury markers (LDH, CK-MB), and decreased apoptosis, reflected by a lower Bax/Bcl-2 ratio. GC preconditioning reduces I/R-induced myocardial injury in rats by activating the PI3K/Akt pathway, reducing oxidative stress, inhibiting apoptosis, and improving cell viability. These results support GC's potential for treating ischemic heart disease and suggest Akt-targeted interventions for myocardial protection. Keywords Ginkgolide C " Ischemia/reperfusion injury " Cardioprotection " PI3K/Akt pathway.
Asunto(s)
Cardiotónicos , Ginkgólidos , Lactonas , Daño por Reperfusión Miocárdica , Fosfatidilinositol 3-Quinasas , Proteínas Proto-Oncogénicas c-akt , Animales , Daño por Reperfusión Miocárdica/tratamiento farmacológico , Daño por Reperfusión Miocárdica/metabolismo , Daño por Reperfusión Miocárdica/fisiopatología , Daño por Reperfusión Miocárdica/patología , Daño por Reperfusión Miocárdica/prevención & control , Ginkgólidos/farmacología , Ginkgólidos/uso terapéutico , Proteínas Proto-Oncogénicas c-akt/metabolismo , Masculino , Lactonas/farmacología , Lactonas/uso terapéutico , Ratas , Transducción de Señal/efectos de los fármacos , Ratas Sprague-Dawley , Fosfatidilinositol 3-Quinasas/metabolismo , Miocitos Cardíacos/efectos de los fármacos , Miocitos Cardíacos/metabolismo , Cardiotónicos/farmacología , Estrés Oxidativo/efectos de los fármacos , Apoptosis/efectos de los fármacosRESUMEN
OBJECTIVE: This experimental study investigates the potential of sodium-glucose cotransporter 2 inhibitor (SGLT2i) dapagliflozin to enhance cardiac hemodynamic and metabolism function following ischemia/reperfusion (I/R) after cold cardioplegic arrest. METHODS: Hearts of adult male C57BL/6â¯J mice were excised and mounted on an isolated working heart system. Cold ischemia (100â¯min) was induced using St. Thomas' #2 cardioplegia, applied every 20â¯min, followed by 30â¯min of reperfusion. Continuous monitoring of cardiac parameters was performed. The experimental groups were the following (1) cold cardioplegic arrest as control (nâ¯=â¯12), (2) dapagliflozin applied to perfusion buffer 30â¯min prior to ischemia (nâ¯=â¯16), (3) arrested hearts receiving dapagliflozin as an adjunct to cardioplegia (nâ¯=â¯13), and (4) no-arrested, no-treated hearts (nâ¯=â¯6). Post-protocol, biopsies of hearts were frozen for subsequent metabolic and RT-qPCR analysis. Additionally, cell culture experiments on HUVECS and cardiac myoblasts were performed. RESULTS: Cold I/R markedly reduced systolic and diastolic function. Notably, dapagliflozin administered before ischemia significantly improved left ventricular recovery by enhancing contractility, aortic flow, and left ventricle diastolic function, whereas adding dapagliflozin only to the cardioplegic solution did not yield similar effects. High-energy phosphate (HEP) measurements revealed a statistically significant increase of myocardial energy charge in hearts treated with dapagliflozin via cardioplegia. These functional and metabolic findings were supported by improved cell viability and higher HEP content in HUVECS and cardiac myoblasts, when pre-treated with SGLT2i. DISCUSSION: Our findings underscore the potential of SGLT2i to improve hemodynamic recovery following cardioplegic arrest, supporting further exploration of dapagliflozin as novel myocardial protective regimen in cardiac surgery.
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Compuestos de Bencidrilo , Glucósidos , Paro Cardíaco Inducido , Corazón , Hemodinámica , Daño por Reperfusión Miocárdica , Inhibidores del Cotransportador de Sodio-Glucosa 2 , Animales , Compuestos de Bencidrilo/farmacología , Masculino , Glucósidos/farmacología , Ratones Endogámicos C57BL , Hemodinámica/efectos de los fármacos , Inhibidores del Cotransportador de Sodio-Glucosa 2/farmacología , Ratones , Paro Cardíaco Inducido/métodos , Daño por Reperfusión Miocárdica/metabolismo , Daño por Reperfusión Miocárdica/fisiopatología , Daño por Reperfusión Miocárdica/prevención & control , Corazón/efectos de los fármacos , Corazón/fisiopatología , Humanos , Función Ventricular Izquierda/efectos de los fármacos , Miocardio/metabolismoRESUMEN
AIMS: Acute myocardial infarction results in significant mortality and chronic heart failure, with reperfusion frequently inducing myocardial ischaemia-reperfusion (IR) injury mediated by infiltrating monocytes and monocyte-derived macrophages (iMacs). The olfactory receptor 2 (Olfr2) is hypothesized to serve as a pivotal inflammatory mediator in this context. This study aimed to elucidate the regulatory role of Olfr2 in mitochondrial homeostasis and inflammation in iMacs during myocardial IR injury. METHODS AND RESULTS: The surface expression of OR6A2 (human ortholog of Olfr2) on monocyte subsets was assessed to determine its association with major adverse cardiovascular events (MACEs) in IR-injured patients. The mechanistic role of Olfr2 in modulating iMacs during myocardial IR injury was investigated using both in vivo and in vitro interventions targeting Olfr2. Elevated OR6A2 levels on human monocytes and octanal, an OR6A2 agonist, were significantly associated with an increased risk of MACEs and correlated with increased oxidative stress and pro-inflammatory responses in patients with IR injury. The genetic ablation of Olfr2 in mice demonstrated significant attenuation of mitochondrial reactive oxygen species (mtROS) and pro-inflammatory cytokine in iMacs, accompanied by diminished immune cell infiltration and reduced cardiomyocyte apoptosis, ultimately ameliorating myocardial IR injury. Mechanistically, Olfr2 activated nuclear receptor subfamily 4 group A member 1 (NR4A1) via cAMP/PKA signalling, promoting dynamin-related protein 1 (Drp1)-mediated mitochondrial fission, which led to mitochondrial mtROS overproduction, mitochondrial membrane potential disruption, mitochondrial apoptosis, and the subsequent release of pro-inflammatory factors through NLRP3 inflammasome activation. Notably, monocyte/macrophage-specific NR4A1 overexpression in Olfr2 knockout mice negated the cardiovascular protection observed during IR injury. CONCLUSION: Elevated OR6A2 expression and octanal levels were significantly associated with an increased risk of MACEs. Our findings identified the Olfr2/cAMP/PKA/NR4A1 axis as a novel signalling pathway contributing to cardiac IR injury by promoting Drp1-mediated mitochondrial fission and subsequent production of pro-inflammatory cytokines.
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Macrófagos , Mitocondrias Cardíacas , Dinámicas Mitocondriales , Monocitos , Infarto del Miocardio , Daño por Reperfusión Miocárdica , Miocitos Cardíacos , Miembro 1 del Grupo A de la Subfamilia 4 de Receptores Nucleares , Receptores Odorantes , Animales , Daño por Reperfusión Miocárdica/metabolismo , Daño por Reperfusión Miocárdica/prevención & control , Daño por Reperfusión Miocárdica/patología , Daño por Reperfusión Miocárdica/genética , Humanos , Dinámicas Mitocondriales/efectos de los fármacos , Monocitos/metabolismo , Monocitos/patología , Monocitos/efectos de los fármacos , Transducción de Señal , Miembro 1 del Grupo A de la Subfamilia 4 de Receptores Nucleares/metabolismo , Miembro 1 del Grupo A de la Subfamilia 4 de Receptores Nucleares/genética , Masculino , Modelos Animales de Enfermedad , Ratones Endogámicos C57BL , Ratones Noqueados , Mitocondrias Cardíacas/metabolismo , Mitocondrias Cardíacas/patología , Mitocondrias Cardíacas/efectos de los fármacos , Dinaminas/metabolismo , Receptores Odorantes/genética , Receptores Odorantes/metabolismo , Receptores Odorantes/agonistas , Receptores Odorantes/deficiencia , Estrés Oxidativo , Células Cultivadas , Especies Reactivas de Oxígeno/metabolismo , Macrófagos/metabolismo , Macrófagos/patología , Miocitos Cardíacos/metabolismo , Miocitos Cardíacos/patología , Infarto del Miocardio/metabolismo , Infarto del Miocardio/patología , Infarto del Miocardio/prevención & control , Infarto del Miocardio/genética , Mediadores de Inflamación/metabolismo , Femenino , Persona de Mediana Edad , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Proteínas Quinasas Dependientes de AMP Cíclico/metabolismo , Inflamasomas/metabolismoRESUMEN
AIMS: Despite advancements in primary percutaneous coronary intervention (PCI), cardiac dysfunction remains a challenge in patients with ST-segment elevation myocardial infarction (STEMI). Although thymosin beta 4 has shown cardioprotective effects in preclinical MI models, its impact on chronic cardiac functional recovery post ischemia/reperfusion (I/R), especially in STEMI, warrants further investigation. This study aims to explore the therapeutic potential of recombinant human thymosin beta 4 (rhTB4) in both murine models subjected to I/R and in subjects with STEMI post-PCI. METHODS AND RESULTS: In C57BL/6J mice, 7-day rhTB4 treatment prevented cardiac dysfunction and fibrosis 28 days post-I/R surgery and significantly reduced plasma NT-proBNP levels at both 1 day and 28 days post-I/R. Similarly, in a permanent ligation model, rhTB4 improved cardiac function and reduced infarct size at 8 weeks post-MI. RNA-seq analysis of I/R heart tissues revealed that rhTB4 modulated the ErbB signaling pathway. In vitro hypoxia/reoxygenation (H/R) models (HL-1, neonatal mouse cardiomyocytes, H9C2) demonstrated that rhTB4 activated the ErbB2/Raf1 signaling pathway, attenuated cardiomyocyte apoptosis and suppressed pro-apoptotic protein Bad expression. The cardioprotective effects of rhTB4 on cardiac function and adverse cardiac remodeling in I/R mice were abolished by ErbB2 inhibition. In a randomized, placebo-controlled, double-blind trial involving 96 STEMI patients, the infarcted areas were significantly reduced in the rhTB4 group, which received the first dose of rhTB4 within 8â h after PCI (n = 43), as compared to the placebo group at the 90-day follow-up. However, the overall differences in infarcted areas were not significantly between the rhTB4 group and the placebo group (n = 96). CONCLUSION: These findings underscore the crucial role of rhTB4 in mitigating cardiac dysfunction in an ErbB2-dependent manner. The clinical relevance of rhTB4 is demonstrated through a randomized controlled trial, emphasizing its translational potential. Further rigorous randomized studies are needed to assess the significance of early rhTB4 use post-myocardial infarction reperfusion.
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Daño por Reperfusión Miocárdica , Miocitos Cardíacos , Intervención Coronaria Percutánea , Infarto del Miocardio con Elevación del ST , Timosina , Función Ventricular Izquierda , Animales , Timosina/farmacología , Humanos , Infarto del Miocardio con Elevación del ST/fisiopatología , Infarto del Miocardio con Elevación del ST/metabolismo , Infarto del Miocardio con Elevación del ST/patología , Infarto del Miocardio con Elevación del ST/tratamiento farmacológico , Ratones Endogámicos C57BL , Modelos Animales de Enfermedad , Masculino , Daño por Reperfusión Miocárdica/fisiopatología , Daño por Reperfusión Miocárdica/metabolismo , Daño por Reperfusión Miocárdica/patología , Daño por Reperfusión Miocárdica/prevención & control , Daño por Reperfusión Miocárdica/tratamiento farmacológico , Transducción de Señal , Recuperación de la Función , Miocitos Cardíacos/efectos de los fármacos , Miocitos Cardíacos/patología , Miocitos Cardíacos/metabolismo , Fibrosis , Proteínas Recombinantes/farmacología , Persona de Mediana Edad , Función Ventricular Izquierda/efectos de los fármacos , Femenino , Factores de Tiempo , Anciano , Remodelación Ventricular/efectos de los fármacos , Apoptosis/efectos de los fármacos , Ratones , Intervención Coronaria Percutánea/efectos adversosRESUMEN
Stopping then restarting the blood flow to the heart can cause ischaemia reperfusion (IR) injury. This can happen during revascularisation following a myocardial infarction and during on pump cardiac surgery using cardioplegic arrest. Despite extensive studies to identify cardioprotective interventions, the myocardium continues to sustain significant injury. Therefore, there is a need to identify agents that can be used during IR. This review focuses on the potential cardioprotective role for acidic amino acids and natural dipeptides using evidence from experimental studies and clinical trials with particular emphasis on their membrane transport. Acidic amino acids are present at high concentration in the heart with a large tissue to plasma concentration gradient, where they are involved in protein synthesis and intermediary metabolism. During cardiac insults they are lost from heart cells but replenishment leads to cardioprotection through energy provision, protection against the production of reactive oxygen species production and improved calcium homeostasis. One important determinant of the intracellular concentration of acidic amino acids and natural dipeptides is membrane transport. The expression and activity of the acidic amino acids transporters EAAT1-3 and the dipeptide transporter, PEPT2 have been demonstrated in membrane vesicles and isolated cardiomyocytes. Improvements in our understanding of these different transport mechanisms should lead to the maximisation of acidic amino acid and natural dipeptide uptake during IR leading to improved cardioprotection.
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Aminoácidos , Cardiotónicos , Dipéptidos , Daño por Reperfusión Miocárdica , Miocardio , Humanos , Dipéptidos/metabolismo , Dipéptidos/uso terapéutico , Dipéptidos/farmacología , Animales , Daño por Reperfusión Miocárdica/metabolismo , Daño por Reperfusión Miocárdica/prevención & control , Daño por Reperfusión Miocárdica/tratamiento farmacológico , Daño por Reperfusión Miocárdica/patología , Aminoácidos/metabolismo , Aminoácidos/uso terapéutico , Cardiotónicos/uso terapéutico , Cardiotónicos/farmacología , Cardiotónicos/metabolismo , Miocardio/metabolismo , Miocardio/patologíaRESUMEN
BACKGROUND: Failing heart is more likely to suffer from myocardial ischemia/reperfusion (I/R) injury. This poses a great challenge for anesthesiologists in managing patients with heart failure during major surgery. Evidence from animal studies suggests that the delta-opioid receptor (DOR) contributes to alleviating acute myocardial injuries. However, little is known regarding the cardioprotective effects of cardiac DOR in patients with chronic heart failure. This study aimed to examine DOR expression in failing hearts and explore how DOR regulates the Janus kinase signal transducer and activator of the transcription-3 (JAK/STAT3) pathway to mediate morphine-induced cardio protection in heart failure. METHODS: We measured the DOR protein levels in human and rat heart tissues with chronic heart failure. To investigate the cardioprotective role of DOR, we administered the DOR-specific antagonist, naltrindole (NTD), and JAK2 inhibitor, AG490, before morphine preconditioning (MPC) in an isolated perfusion model of myocardial I/R injury in postinfarcted failing rat heart. We examined the infarct size, cardiac enzymes, cardiac function, cardiomyocyte apoptosis, apoptosis-related proteins, and STAT3 phosphorylation in the heart. RESULTS: The protein levels of DOR were significantly elevated in the myocardial tissues of humans and rats with chronic heart failure, by 1.4-fold (mean difference 0.41; 95% confidence interval [CI], 0.04-0.78; P = .032) and 2.3-fold (mean difference 1.26; 95% CI, 0.25-2.28; P = .009), respectively, compared to control tissues. Disease severity positively correlated with DOR expression (human: R2 = 0.316, P = .004; rat: R2 = 0.871, P = .021). Blocking DOR substantially reversed the cardioprotective effects of MPC in postinfarcted rat hearts, increasing the mean (standard deviation) percentage of infarct size from 15.0 (3.9)% to 30.8 (7.7)% (P < .001). Similarly, AG490 inhibited MPC restoration of cardiomyocyte apoptosis (33.3 [4.2]% vs 16.6 [3.4]%; P < .001). Both NTD and AG490 markedly suppressed STAT3 phosphorylation by 60.1% (mean difference 0.60; 95% CI, 0.27-0.93; P = .002) and 44.1% (mean difference 0.44; 95% CI, 0.06-0.83; P = .027), respectively, and also lowered the Bcl-2/Bax ratio by 85.5% (mean difference 0.86; 95% CI, 0.28-1.43; P = .006) and 68.2% (mean difference 0.68; 95% CI, 0.51-0.85; P < .001) respectively in heart tissues at the end of reperfusion. CONCLUSIONS: DOR protein levels increased in failing hearts of both humans and rats. Blocking cardiac DOR selectively reduced morphine-induced cardio protection by inhibiting the JAK2/STAT3 pathway. These findings indicate that cardiac DOR is a potential therapeutic target for protecting against heart failure due to I/R injury.
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Cardiotónicos , Insuficiencia Cardíaca , Janus Quinasa 2 , Morfina , Daño por Reperfusión Miocárdica , Receptores Opioides delta , Factor de Transcripción STAT3 , Animales , Factor de Transcripción STAT3/metabolismo , Janus Quinasa 2/metabolismo , Janus Quinasa 2/antagonistas & inhibidores , Morfina/farmacología , Morfina/administración & dosificación , Receptores Opioides delta/metabolismo , Receptores Opioides delta/antagonistas & inhibidores , Receptores Opioides delta/agonistas , Humanos , Insuficiencia Cardíaca/patología , Insuficiencia Cardíaca/tratamiento farmacológico , Insuficiencia Cardíaca/enzimología , Insuficiencia Cardíaca/fisiopatología , Insuficiencia Cardíaca/metabolismo , Masculino , Transducción de Señal/efectos de los fármacos , Transducción de Señal/fisiología , Ratas Sprague-Dawley , Ratas , Persona de Mediana Edad , Daño por Reperfusión Miocárdica/prevención & control , Daño por Reperfusión Miocárdica/patología , Daño por Reperfusión Miocárdica/enzimología , Cardiotónicos/farmacología , Apoptosis/efectos de los fármacos , Anciano , Tirfostinos/farmacología , Modelos Animales de Enfermedad , Analgésicos Opioides/farmacología , Miocitos Cardíacos/efectos de los fármacos , Miocitos Cardíacos/patología , Fosforilación , FemeninoRESUMEN
Myocardial ischemia-reperfusion injury (MIRI) endures as a substantial impediment to the management of cardiovascular disease. The pathophysiology of MIRI is complex, involving oxidative stress, calcium overload, inflammation, and apoptosis. The NRG1/ErbB4 signaling pathway has been implicated in modulating oxidative stress responses in the heart, potentially reducing cellular damage caused by free radicals. This study aimed to elucidate the molecular mechanisms by which the NRG1/ErbB4 pathway confers protection in a rat model of myocardial ischemia-reperfusion (MIRI) injury. A rat MIRI model was established, involving 30 adult male Sprague-Dawley rats randomly assigned to five groups: control, sham operation, MIRI, recombinant NRG1, and AG1478. NRG1 concentrations in heart tissues were measured by ELISA, while p-ErbB4 and total ErbB4 protein levels were assessed by Western blot. Cardiomyocyte apoptosis and reactive oxygen species (ROS) levels were evaluated by flow cytometry.In the MIRI group, NRG1 levels were significantly reduced compared to the control and sham operation groups (p < 0.05). Recombinant NRG1 treatment led to a marked increase in NRG1 levels in heart tissue, along with an enhanced p-ErbB4/ErbB4 expression ratio (p < 0.05). The AG1478 group exhibited lower NRG1 levels and a reduced p-ErbB4/ErbB4 ratio compared to the recombinant NRG1 group (p < 0.05). In terms of apoptosis and ROS levels, the MIRI group showed significantly higher rates than the control and sham operation groups (p < 0.05). Recombinant NRG1 treatment significantly reduced myocardial cell apoptosis and ROS levels (p < 0.05), while AG1478 also attenuated these effects, but to a lesser extent. The NRG1/ErbB4 signaling pathway plays a pivotal protective role in MIRI in rats. Supplementation with exogenous NRG1 effectively elevates NRG1 levels in the heart, activates the ErbB4 signaling pathway, reduces myocardial cell apoptosis, and decreases ROS levels, thereby mitigating MIRI-induced damage.
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Receptores ErbB , Daño por Reperfusión Miocárdica , Neurregulina-1 , Animales , Neurregulina-1/metabolismo , Neurregulina-1/farmacología , Neurregulina-1/administración & dosificación , Masculino , Ratas Sprague-Dawley , Receptor ErbB-4/metabolismo , Daño por Reperfusión Miocárdica/metabolismo , Daño por Reperfusión Miocárdica/prevención & control , Daño por Reperfusión Miocárdica/patología , Ratas , Transducción de Señal/fisiología , Especies Reactivas de Oxígeno/metabolismo , Apoptosis/efectos de los fármacos , Miocitos Cardíacos/metabolismo , Miocitos Cardíacos/patología , Receptores ErbB/metabolismo , Tirfostinos/farmacología , Proteínas Recombinantes/administración & dosificación , Proteínas Recombinantes/farmacologíaRESUMEN
Reperfusion arrhythmia (RA) poses a significant risk to the prognosis of patients undergoing extracorporeal circulation cardiac surgery, and its occurrence closely linked to disturbances in myocardial electrical conduction. Our prior research indicated that ischemia-reperfusion (I/R) affects the localization of connexin 43 (Cx43) at the intercalated discs (IDs) by inducing depolymerization of microtubules, resulting in myocardial electrical conduction abnormalities in rats. Although sevoflurane preconditioning (SPC) has demonstrated considerable protective effects on ischemic myocardium, the underlying mechanisms remain inadequately understood. This prompted us to study the effect of SPC on the stability of microtubules in I/R myocardium to explore the protective mechanism of SPC based on microtubule cytoskeleton. Western blotting, immunofluorescence colocalization, and ex vivo cardiac electrophysiological mapping demonstrated that SPC attenuated myocardial I/R-induced microtubule depolymerization, recovered the distribution of Cx43 at the IDs, and improved myocardial electrical conduction and RA score in rats. Furthermore, the microtubule depolymerization agent nocodazole abolished the protective effect of SPC on microtubules and significantly inhibited the ameliorative effects of sevoflurane on Cx43 localization and electrical conduction. These findings substantiate that the ability of SPC to restore the localization of Cx43 and electrophysiological functions in I/R myocardium depends on microtubule stability. In conclusion, our study shows that SPC ensures the targeted transport of Cx43 to the IDs by stabilizing microtubule structure, thereby ameliorating myocardial electrical conduction and RA after I/R.
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Conexina 43 , Éteres Metílicos , Microtúbulos , Daño por Reperfusión Miocárdica , Miocardio , Sevoflurano , Conexina 43/metabolismo , Animales , Microtúbulos/efectos de los fármacos , Microtúbulos/metabolismo , Masculino , Daño por Reperfusión Miocárdica/metabolismo , Daño por Reperfusión Miocárdica/fisiopatología , Daño por Reperfusión Miocárdica/prevención & control , Ratas Sprague-Dawley , Ratas , Miocardio/metabolismo , Miocardio/patología , Éteres Metílicos/farmacología , Sistema de Conducción Cardíaco/efectos de los fármacosRESUMEN
BACKGROUND: Hyperlipidemia significantly exacerbates myocardial ischemia-reperfusion (I/R) injurfy through lipid metabolic dysfunction and lipotoxicity. Current evidence suggests that lipid droplet accumulation and impaired lipophagy represent critical pathological mechanisms underlying cardiac dysfunction in hyperlipidemic conditions. This study investigated the cardioprotective effects of asiaticoside (AS) against myocardial I/R injury in hyperlipidemic mice and elucidated its underlying mechanisms, emphasizing the AMPK/ORP8-mediated lipophagy pathway. METHODS: Hyperlipidemic C57BL/6 mice were established using high-fat diet feeding and subjected to myocardial I/R injury. Mice received AS (12.5, 25, or 50 mg/kg) treatment for 4 weeks prior to surgery. In vitro experiments involved H9C2 cardiomyocytes treated with palmitic acid followed by hypoxia/reoxygenation. The role of AMPK/ORP8 signaling was evaluated using pharmacological modulators [AMPK activator (A-769662) and AMPK inhibitor (Compound C)] and genetic manipulation (ORP8 siRNA knockdown). RESULTS: AS dose-dependently improved cardiac function parameters, reduced myocardial infarct size (LVEF and LVFS) and decreased triglyceride and cardiac injury biomarkers (cTnI, LDH, CK-MB) in hyperlipidemic I/R mice. Treatment with AS significantly reduced cardiac lipid accumulation and triglyceride content while enhancing lipophagy markers (LC3B-II and Beclin-1) and reducing p62 levels. Mechanistically, AS activated AMPK phosphorylation and upregulated ORP8 expression, which was accompanied by enhanced lipophagy flux. In H9C2 cells, AS protected against palmitic acid-induced lipotoxicity and H/R injury through AMPK/ORP8-dependent lipophagy activation. AMPK inhibition (Compound C) or ORP8 knockdown significantly attenuated AS's protective effects, while AMPK activation (A-769,662) potentiated these benefits, which were reversed to some extent by ORP8 silencing. CONCLUSIONS: This study demonstrates that AS mitigates myocardial I/R injury in hyperlipidemic conditions by promoting lipophagy through the AMPK/ORP8 signaling axis. The AMPK/ORP8-lipophagy pathway represents a novel therapeutic target for metabolic cardiovascular diseases, and AS emerges as a promising cardioprotective agent with significant translational potential.
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Proteínas Quinasas Activadas por AMP , Cardiotónicos , Hiperlipidemias , Daño por Reperfusión Miocárdica , Triterpenos , Animales , Hiperlipidemias/complicaciones , Hiperlipidemias/tratamiento farmacológico , Hiperlipidemias/metabolismo , Ratones Endogámicos C57BL , Daño por Reperfusión Miocárdica/tratamiento farmacológico , Daño por Reperfusión Miocárdica/prevención & control , Daño por Reperfusión Miocárdica/metabolismo , Masculino , Proteínas Quinasas Activadas por AMP/metabolismo , Cardiotónicos/farmacología , Ratones , Autofagia/efectos de los fármacos , Miocitos Cardíacos/efectos de los fármacos , Triterpenos/farmacología , Dieta Alta en Grasa , Transducción de Señal/efectos de los fármacos , Línea Celular , RatasRESUMEN
INTRODUCTION: Cardioplegias are routinely used in cardiac surgery to protect the heart from ischemia-reperfusion injury. The choice of cardioplegia depends on the surgeon's clinical expertise. No clear data demonstrate the redox-protective superiority of one cardioplegia over another. We aim to evaluate redox status and signaling assessments in coronary sinus blood samples for the different cardioplegias. METHODS: Our study included patients undergoing coronary artery bypass and isolated valve surgery. We compared blood and del Nido cardioplegia solutions. During the preoperative period, blood samples were collected from the coronary sinus both preand post-aortic cross-clamping. We also assessed redox system biomarkers and transcription factors related to the antioxidant system using spectrophotometric and immunochemical methods. RESULTS: In valve patient groups that received both cardioplegia solutions, post-cross-clamping protein carbonyl levels were significantly lower compared to pre-cross-clamping values. For the levels of antioxidant system parameters, except for catalase and superoxide dismutase, no significant difference was observed for del Nido cardioplegia. Increased antioxidant enzyme levels highlight the importance of these enzymes in eliminating the higher hydroperoxide load. Regulatory proteins involved in redox signaling did not show significant variations except for Kelch-like ECH-associated protein 1 and peroxisome proliferator-activated receptor-gamma coactivator-1 alpha for cardioplegias. CONCLUSION: Current results indicate that del Nido cardioplegia effectively protects myocardial redox status. Given these findings, despite concerns regarding its use in clinical practice, particularly in valve surgery compared to coronary artery bypass surgery, del Nido cardioplegia may provide effective myocardial protection in both coronary artery bypass and heart valve surgeries.
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Soluciones Cardiopléjicas , Paro Cardíaco Inducido , Daño por Reperfusión Miocárdica , Factores de Transcripción , Humanos , Biomarcadores/sangre , Masculino , Paro Cardíaco Inducido/métodos , Oxidación-Reducción/efectos de los fármacos , Femenino , Soluciones Cardiopléjicas/farmacología , Persona de Mediana Edad , Puente de Arteria Coronaria/métodos , Factores de Transcripción/sangre , Factores de Transcripción/análisis , Anciano , Daño por Reperfusión Miocárdica/prevención & control , Transducción de Señal , Seno Coronario , AntioxidantesRESUMEN
Objective: Annexin-A1 (ANXA1) participates in regulating ferroptosis. Ferroptosis is involved in myocardial ischemia-reperfusion injury (MIRI). However, effect and mechanism of ANXA1 in ferroptosis after MIRI remain unclear. Materials and Methods: Gene expression omnibus database was used to screen differentially expressed genes (DEGs), and R package was used to visualize the volcano map and heat map of DEGs in MIRI. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes analyses were used to analyze the related signaling pathways and functions of DEGs. FerrDb database was used to find genes related to ferroptosis. H9c2 cells were cultured under hypoxia/reoxygenation (H/R). Quantitative real-time polymerase chain reaction and western blot detected mRNA and protein expression of relevant genes. Mitochondrial membrane potential was determined by JC-1. Cell viability was assessed using CCK-8. Levels of Fe2+, glutathione, malondialdehyde, and reactive oxygen species (ROS) were detected by kits. Results: Genes such as ANXA1 were highly expressed in MIRI compared to normal tissues. Hypoxic cardiomyocytes had enhanced viability after knocking down ANXA1. ANXA1 depletion improved ferroptosis and altered mitochondrial functioning in cardiomyocytes under H/R condition. Ferroptosis inducer Erastin reversed this phenomenon. Phosphorylation levels of c-Jun N-terminal kinase, P38, and Raf in H9c2 cells after ANXA1 silencing were increased. With treatment of RAF inhibitor Sorafenib, above results were reversed. ANXA1 depletion alleviated ferroptosis and mitochondrial damage by activating RAS/Raf/mitogen-activated protein kinase (MAPK) pathway in H/R-induced cardiomyocytes. Conclusions: ANXA1 knockdown reduces cardiomyocyte ferroptosis after MIRI by regulating RAS/Raf/MAPK signaling pathway, which provides new therapeutic targets for MIRI treatment.
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Anexina A1 , Ferroptosis , Daño por Reperfusión Miocárdica , Miocitos Cardíacos , Animales , Ratones , Ratas , Anexina A1/genética , Anexina A1/metabolismo , Línea Celular , Conjuntos de Datos como Asunto , Ferroptosis/efectos de los fármacos , Ferroptosis/genética , Perfilación de la Expresión Génica , Técnicas de Silenciamiento del Gen , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Sistema de Señalización de MAP Quinasas/genética , Potencial de la Membrana Mitocondrial , Mitocondrias/efectos de los fármacos , Mitocondrias/metabolismo , Mitocondrias/patología , Daño por Reperfusión Miocárdica/tratamiento farmacológico , Daño por Reperfusión Miocárdica/genética , Daño por Reperfusión Miocárdica/patología , Daño por Reperfusión Miocárdica/prevención & control , Miocardio/citología , Miocardio/patología , Miocitos Cardíacos/efectos de los fármacos , Miocitos Cardíacos/metabolismo , Miocitos Cardíacos/patología , Quinasas raf/antagonistas & inhibidores , Quinasas raf/metabolismo , Proteínas ras/metabolismo , Sorafenib/farmacología , PiperazinasRESUMEN
BACKGROUND: Pharmacological postconditioning (PostC) is an emerging and safer cardioprotective strategy against myocardial ischemia-reperfusion injury (MIRI), involving the administration of therapeutic agents at the onset of reperfusion. Activation of the cannabinoid B2 receptor plays a crucial role in cardioprotection across various cardiovascular pathologies. AM-1241, a selective cannabinoid B2 receptor agonist, exhibits multiple pharmacological properties, including anti-oxidant, anti-inflammatory, anti-apoptotic, and cardioprotective effects. METHODOLOGY: MIRI was induced in isolated Wistar rat hearts using the Langendorff Power Lab system by subjecting them to 30 min of global ischemia followed by 120 min of reperfusion. AM-1241 PostC (3 µM and 6 µM) was given through four cycles of 30-s ischemia and reperfusion (stoppage/free flow of the drug, respectively) before the prolonged 120-min reperfusion. RESULTS: MIRI-induced myocardial damage was evident through increased infarct size, elevated cardiac injury markers such as Lactate dehydrogenase-1 (LDH-1), Creatine kinase-MB (CK-MB), Cardiac troponin-I (C-tPn-I), impaired hemodynamic parameters including heart rate (HR), coronary flow rate (CFR), left ventricular developed pressure (LVDP), rate pressure product (RPP), +dp/dtmax, -dp/dtmin, and biochemical alterations such as increased thiobarbituric acid reactive species (TBARS), tumor necrosis factor-α (TNF-α), transforming growth factor- ß (TGF-ß), Bax, caspase-3; decreased glutathione reductase (GSH) and catalase. AM-1241 PostC significantly reduced infarct size and cardiac injury markers while improving hemodynamic and biochemical parameters. However, prior PostC of AM-630 (selective cannabinoid B2 receptor antagonist; 25 µM and 50 µM) and BML-275, an AMP activated protein kinase (AMPK) mediated autophagy inhibitor; 10 µM and 20 µM substantially abolished the cardioprotective effects of AM-1241 PostC. CONCLUSION: These findings suggest that AM-1241 PostC exerts cardioprotective effects in MIRI through modulation of inflammatory, fibrosis, apoptotic and AMPK-mediated autophagy pathways.
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Cannabinoides , Cardiotónicos , Daño por Reperfusión Miocárdica , Apoptosis/efectos de los fármacos , Apoptosis/fisiología , Fibrosis , Autofagia/fisiología , Daño por Reperfusión Miocárdica/tratamiento farmacológico , Daño por Reperfusión Miocárdica/prevención & control , Cardiotónicos/farmacología , Cardiotónicos/uso terapéutico , Masculino , Femenino , Animales , Ratas , Ratas Wistar , Inflamación/tratamiento farmacológico , Miocardio/inmunología , Miocardio/metabolismo , Receptor Cannabinoide CB2/metabolismo , Agonistas de Receptores de Cannabinoides/farmacología , Agonistas de Receptores de Cannabinoides/uso terapéutico , Transducción de Señal/efectos de los fármacos , Transducción de Señal/fisiología , Quinasas de la Proteína-Quinasa Activada por el AMP/metabolismo , Cannabinoides/farmacología , Cannabinoides/uso terapéuticoRESUMEN
Ischemiareperfusion is a common cause of myocardial injury, and there are currently no clinically approved drugs for its treatment. Oxidative stress caused by mitochondrial ROS is the main cause of cell damage in ischemiareperfusion (I/R). Therefore, exploring methods to reduce mitochondrial ROS during I/R is essential. Here, we investigated the effects of a low concentration of the uncoupler FCCP on hypoxia/reoxygenation (H/R) injury in myocardial cells. An in vitro myocardial I/R model was constructed by using sodium sulfite (Na2SO3) to induce hypoxia and then reoxygenation. We found that after hypoxia, treatment with 5â¯nM FCCP induced the expression of UCP1, which uncouples mitochondria, thereby decreasing ATP production, ROS levels, and mitophagy, ultimately reducing myocardial injury. In vivo experiments further revealed that 1â¯mg/kg body weight FCCP has a protective effect against myocardial I/R injury. These data indicated that mildly uncoupling mitochondria via a low concentration of FCCP attenuated I/R-triggered cardiac injury.
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Carbonil Cianuro p-Trifluorometoxifenil Hidrazona , Mitocondrias Cardíacas , Daño por Reperfusión Miocárdica , Miocitos Cardíacos , Desacopladores , Animales , Daño por Reperfusión Miocárdica/metabolismo , Daño por Reperfusión Miocárdica/patología , Daño por Reperfusión Miocárdica/prevención & control , Especies Reactivas de Oxígeno/metabolismo , Miocitos Cardíacos/efectos de los fármacos , Miocitos Cardíacos/metabolismo , Miocitos Cardíacos/patología , Masculino , Mitocondrias Cardíacas/efectos de los fármacos , Mitocondrias Cardíacas/metabolismo , Mitocondrias Cardíacas/patología , Carbonil Cianuro p-Trifluorometoxifenil Hidrazona/farmacología , Hipoxia de la Célula/efectos de los fármacos , Ratones , Desacopladores/farmacología , Estrés Oxidativo/efectos de los fármacos , Proteína Desacopladora 1/metabolismo , Mitofagia/efectos de los fármacos , Adenosina Trifosfato/metabolismo , Ratones Endogámicos C57BLRESUMEN
OBJECTIVES: Myocardial protection is crucial for recovery after cardiac surgery. Despite older patients with more comorbidities, most surgeons still use potassium-induced depolarized arrest with cardioplegic solutions from more than 50 years ago. However, this can cause calcium overload, mitochondrial injury and energy depletion. We tested a new polarizing cardioplegia (STH-pol) versus traditional depolarized arrest (STH-control) in an ischaemia/reperfusion model of hearts with reduced ejection fraction. METHODS: Myocardial infarction (MI) was induced by permanent ligation of the left anterior descending artery in male Sprague-Dawley rats. Six weeks post-MI, the hearts were isolated and perfused on an erythrocyte-perfused working heart system. Cold ischaemic arrest (4°C, 60 min) was induced by STH-pol (n = 10) or STH-control (n = 8), followed by reperfusion (37°C, 45 min). Haemodynamic variables were recorded throughout reperfusion, and tissue samples were taken at the end of reperfusion to analyse high-energy phosphates (HEP). RESULTS: Administration of STH-pol resulted in comparable haemodynamic recovery after 60 min ischaemia as STH-control induced cardioplegic arrest (left ventricular systolic pressure: 109.1 ± 3.4 vs 103.3 ± 3.6). Similarly, HEP levels did not differ between the two groups (ATP: 2.5 ± 0.4 nmol/mg protein and 2.2 ± 0.9 nmol/mg). CONCLUSIONS: Polarizing cardioplegic arrest did not lead to a statistically significant difference in myocardial protection as compared to a clinically relevant, standard cold blood cardioplegia in rat hearts with chronic MI.
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Soluciones Cardiopléjicas , Paro Cardíaco Inducido , Infarto del Miocardio , Daño por Reperfusión Miocárdica , Volumen Sistólico , Función Ventricular Izquierda , Animales , Paro Cardíaco Inducido/métodos , Paro Cardíaco Inducido/efectos adversos , Masculino , Ratas Sprague-Dawley , Soluciones Cardiopléjicas/farmacología , Infarto del Miocardio/fisiopatología , Infarto del Miocardio/metabolismo , Daño por Reperfusión Miocárdica/prevención & control , Daño por Reperfusión Miocárdica/fisiopatología , Daño por Reperfusión Miocárdica/metabolismo , Daño por Reperfusión Miocárdica/etiología , Modelos Animales de Enfermedad , Ratas , Función Ventricular Izquierda/efectos de los fármacos , Miocardio/metabolismo , Miocardio/patología , Hemodinámica/efectos de los fármacos , Cloruro de Potasio , Bicarbonatos , Cloruro de Calcio , Cloruro de Sodio , MagnesioRESUMEN
Evidence suggests that autophagy plays a crucial role in myocardial ischemia/reperfusion (I/R) injury. Renalase (RNLS) plays a protective role in cardiovascular diseases, including myocardial ischemia/reperfusion injury (MI/RI). Therefore, we used both in vivo and in vitro I/R models to monitor autophagy in the cardiomyocytes by exposing H9c2 cells to oxygen and glucose deprivation/reoxygenation (OGD/R) and subjecting rats to I/R, respectively. Wistar rats (male, 8 weeks, n = 27) were randomly divided into a Sham group, I/R + Vehicle group, and I/R + recombinant renalase protein (Re-RNLS) group. An MI/RI model was induced by left anterior descending artery ligation/release, TTC-Evans blue double staining was utilized to evaluate the area of myocardial infarction and ischemic area 24 hours after the operation, and plasma lactate dehydrogenase (LDH) and creatine kinase (CK) activities, plasma RNLS, epinephrine, and dopamine concentrations were detected by ELISA. H9c2 cells were treated with OGD/R serum and pretreated with Re-RNLS, chloroquine (CQ), and MK2206. Western blotting detected protein expression changes in LC3 autophagosomes were monitored by immunofluorescence, and cell death was determined by flow cytometry. The level of RNLS in serum was significantly increased during MI/RI in rats, and Re-RNLS significantly reduced myocardial infarct size after I/R surgery (24.6 ± 4.1% versus 11.2 ± 1.2%). RNLS deficiency aggravated the OGD/R injury in H9c2 cells, and Re-RNLS inhibited the expression of LC3 and Beclin-1 in H9c2 cells subjected to OGD/R, thereby reducing autophagosome formation. Re-RNLS activated the Akt/mTOR pathway of H9c2 cells to play a cytoprotective role. RNLS protects against MI/RI injury by inhibiting autophagy through activating the Akt/mTOR pathway.
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Autofagia , Daño por Reperfusión Miocárdica , Monoéster Fosfórico Hidrolasas , Proteínas Proto-Oncogénicas c-akt , Serina-Treonina Quinasas TOR , Animales , Daño por Reperfusión Miocárdica/metabolismo , Daño por Reperfusión Miocárdica/patología , Daño por Reperfusión Miocárdica/prevención & control , Daño por Reperfusión Miocárdica/tratamiento farmacológico , Autofagia/efectos de los fármacos , Masculino , Ratas , Serina-Treonina Quinasas TOR/metabolismo , Ratas Wistar , Miocitos Cardíacos/metabolismo , Miocitos Cardíacos/efectos de los fármacos , Miocitos Cardíacos/patología , Proteínas Proto-Oncogénicas c-akt/metabolismo , Transducción de Señal/efectos de los fármacos , Modelos Animales de Enfermedad , Monoéster Fosfórico Hidrolasas/farmacologíaRESUMEN
Tea polyphenols have been shown to prevent cardiovascular disease through antioxidant and anti-inflammatory mechanisms. Theogallin, a phenolic acid derived from gallic and quinic acids, is a unique polyphenolic compound found in teas. Its preventive effect on myocardial ischemia-reperfusion (I/R) injury remains unclear. This study utilized an ex vivo model of I/R injury in isolated rat hearts through Langendorff perfusion and an in vivo myocardial injury model through left anterior descending coronary artery (LAD) ligation. In isolated hearts, theogallin pretreatment at both 100 and 200 ng/mL significantly improved hemodynamic parameters following I/R injury. Additionally, in vivo, pretreatment with 20 mg/kg theogallin in LAD ligation rats for 7 days significantly enhanced myocardial contractile function, reduced the release of myocardial enzymes, and decreased infarct size and fibrosis in LAD ligation rats. These protective effects may be attributed to theogallin's ability to inhibit the expression of interleukin-17 (IL-17) receptor A, transcription factor Jun-B (JUNB), Fos-related antigen 1 (FRA1), and matrix metalloproteinase 9 (MMP9) in the IL-17 signaling pathway. These findings suggest that theogallin exerts cardioprotective effects by downregulating the IL-17 signaling pathway in rats with myocardial I/R injury.
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Cardiotónicos , Ácido Gálico , Interleucina-17 , Daño por Reperfusión Miocárdica , Transducción de Señal , Animales , Daño por Reperfusión Miocárdica/genética , Daño por Reperfusión Miocárdica/metabolismo , Daño por Reperfusión Miocárdica/prevención & control , Daño por Reperfusión Miocárdica/tratamiento farmacológico , Masculino , Interleucina-17/genética , Interleucina-17/metabolismo , Transducción de Señal/efectos de los fármacos , Ratas , Ratas Sprague-Dawley , Humanos , Cardiotónicos/administración & dosificación , Ácido Gálico/administración & dosificación , Ácido Gálico/análogos & derivados , Metaloproteinasa 9 de la Matriz/genética , Metaloproteinasa 9 de la Matriz/metabolismo , Camellia sinensis/químicaRESUMEN
BACKGROUND: Myocardial ischemia/reperfusion (I/R) injury is a significant complication following acute myocardial infarction (AMI) and lacks effective therapies. The involvement of gut microbiota in regulating ferroptosis during myocardial I/R injury has not been thoroughly explored. This study aimed to investigate the effect of Lactobacillus on myocardial I/R injury and explore its potential mechanisms. METHODS: One hundred fifty eight patients with ST-elevation myocardial infarction (STEMI) were enrolled in our prospective observational study. The correlations between Lactobacillus levels and myocardial injury markers, inflammatory factors, oxidative stress, and ferroptosis were evaluated. Furthermore, 30 rats were treated with Lactobacillus or vehicle control for 4 weeks, followed by myocardial I/R surgery. The protective effects of Lactobacillus against I/R injury were assessed by quantifying myocardial apoptosis, inflammation, oxidative stress, and ferroptosis. In addition, the above results were verified in vitro. The signaling pathways were investigated through the knockdown and overexpression of sirtuin 1 (Sirt1) and nuclear factor erythroid 2-related factor 2 (Nrf2). RESULTS: In clinical study, Lactobacillus levels were significantly negatively correlated with myocardial injury markers, inflammatory factors, and malondialdehyde (MDA), but positively correlated with glutathione (GSH). In rats, Lactobacillus decreased the levels of myocardial injury markers, reduced the size of the myocardial infarction area, ameliorated the disordered myocardial cell arrangement, and improved cardiac function. In both in vivo and in vitro studies, Lactobacillus inhibited cardiomyocyte apoptosis by upregulated B-cell lymphoma-2 (Bcl-2), downregulated Bcl-2 associated X (Bax), and caspase-3. Furthermore, Lactobacillus decreased inflammatory factors, MDA, reactive oxygen species (ROS) levels, and increased superoxide dismutase (SOD) activity. For ferroptosis, Lactobacillus upregulated the expression of glutathione peroxidase 4 (GPX4) and downregulated the expressions of acyl-CoA synthetase long-chain family member 4 (ACSL4) and transferrin receptor protein 1 (TfR1). Finally, knockdown and overexpression of Sirt1 and Nrf2 in vitro demonstrated that Lactobacillus exerted the effect by upregulating the Sirt1/Nrf2/HO-1 pathway. CONCLUSIONS: Our findings reveals that Lactobacillus protects against myocardial I/R injury by inhibiting apoptosis, inflammation, oxidative stress, and ferroptosis through the Sirt1/Nrf2/HO-1 signaling axis, suggesting a novel probiotic-based therapeutic potential for I/R injury.