Effect and mechanism of Poria cocos polysaccharides on myocardial cell apoptosis in rats with myocardial ischemia-reperfusion injury by regulating Rho-ROCK signaling pathway / 中国中药杂志

This study aimed to investigate the effect and underlying mechanism of Poria cocos polysaccharides(PCP) on myocardial cell apoptosis in the rat model of myocardial ischemia-reperfusion injury(MI/RI). Male SPF-grade SD rats were randomly divided into a sham group(saline), a model group(saline), low-and high-dose PCP groups(100 and 200 mg·kg~(-1)), and a fasudil group(10 mg·kg~(-1)), with 16 rats in each group. Except for the sham group, the other four groups underwent left anterior descending coronary artery ligation for 30 min followed by reperfusion for 2 h to establish the MI/RI model. The myocardial infarct area was assessed by TTC staining. Histological changes were observed through HE staining. Myocardial cell apoptosis was evaluated using TUNEL staining. Serum lactate dehydrogenase(LDH), creatine kinase MB(CK-MB), interleukin-1β(IL-1β) and IL-18 levels, myocardial superoxide dismutase(SOD) activity and malondialdehyde(MDA) levels were detected by ELISA. Protein expression of B-cell lymphoma 2(Bcl-2), Bcl-2 associated X protein(Bax), cleaved caspase-3, Ras homolog gene A(RhoA), myosin phosphatase target subunit 1(MYPT-1), phosphorylated MYPT-1(p-MYPT-1), and Rho-associated coiled-coil forming kinase 1(ROCK 1) were measured by Western blot. Pathological staining of myocardial tissue revealed that in the model group, there was focal necrosis of myocardial tissue, myocardial cell swelling, unclear boundaries, and neutrophil infiltration. These pathological changes were alleviated in the low-and high-dose PCP groups and the fasudil group. Compared with the model group, the low-and high-dose PCP groups and the fasudil group showed significantly reduced myocardial infarct area and myocardial cell apoptosis rate. Compared with the sham group, the model group exhibited elevated serum LDH, CK-MB, IL-1β and IL-18 levels, increased MDA levels, relative protein expression of Bax, cleaved caspase-3, RhoA, ROCK1 and p-MYPT-1, and decreased myocardial SOD levels and Bcl-2 protein expression. Compared with the model group, the PCP groups and the fasudil group showed lowered serum LDH, CK-MB, IL-1β and IL-18 levels, decreased MDA levels, relative protein expression of Bax, cleaved caspase-3, RhoA, ROCK1 and p-MYPT-1, and increased myocardial SOD levels and Bcl-2 protein expression. PCP exhibited a certain preventive effect on myocardial tissue pathological damage and myocardial cell apoptosis in MI/RI rats, possibly related to the inhibition of the Rho-ROCK signaling pathway activation, thereby reducing oxidative stress and inflammatory responses.

Huoxin Pill Reduces Myocardial Ischemia Reperfusion Injury in Rats via TLR4/NFκB/NLRP3 Signaling Pathway / 中国结合医学杂志

OBJECTIVE@#To explore the protective effect of Huoxin Pill (HXP) on acute myocardial ischemia-reperfusion (MIRI) injury in rats.@*METHODS@#Seventy-five adult SD rats were divided into the sham-operated group, model group, positive drug group (diltiazem hydrochloride, DH), high dose group (24 mg/kg, HXP-H) and low dose group (12 mg/kg, HXP-L) of Huoxin Pill (n=15 for every group) according to the complete randomization method. After 1 week of intragastric administration, the left anterior descending coronary artery of the rat's heart was ligated for 45 min and reperfused for 3 h. Serum was separated and the levels of creatine kinase (CK), creatine kinase isoenzyme (CK-MB) and lactate dehydrogenase (LDH), superoxide dismutase (SOD), and malondialdehyde (MDA), hypersensitive C-reactive protein (hs-CRP) and interleukin-1β (IL-1β) were measured. Myocardial ischemia rate, myocardial infarction rate and myocardial no-reflow rate were determined by staining with Evans blue and 2,3,5-triphenyltetrazolium chloride (TTC). Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) and Bioinformatics Analysis Tool for Molecular mechANism of Traditional Chinese Medicine (BATMAN) databases were used to screen for possible active compounds of HXP and their potential therapeutic targets; the results of anti-inflammatory genes associated with MIRI were obtained from GeneCards, Drugbank, Online Mendelian Inheritance in Man (OMIM), and Therapeutic Target Datebase (TTD) databases was performed; Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment were used to analyze the intersected targets; molecular docking was performed using AutoDock Tools. Western blot was used to detect the protein expression of Toll-like receptor 4 (TLR4)/nuclear factor kappa-B (NFκB)/NOD-like receptor protein 3 (NLRP3).@*RESULTS@#Compared with the model group, all doses of HXP significantly reduced the levels of LDH, CK and CK-MB (P<0.05, P<0.01); HXP significantly increased serum activity of SOD (P<0.05, P<0.01); all doses of HXP significantly reduced the levels of hs-CRP and IL-1β (P<0.05, P<0.01) and the myocardial infarction rate and myocardial no-reflow rate (P<0.01). GO enrichment analysis mainly involved positive regulation of gene expression, extracellular space and identical protein binding, KEGG pathway enrichment mainly involved PI3K-Akt signaling pathway and lipid and atherosclerosis. Molecular docking results showed that kaempferol and luteolin had a better affinity with TLR4, NFκB and NLRP3 molecules. The protein expressions of TLR4, NFκB and NLRP3 were reduced in the HXP group (P<0.01).@*CONCLUSIONS@#HXP has a significant protective effect on myocardial ischemia-reperfusion injury in rats, and its effect may be related to the inhibition of redox response and reduction of the inflammatory response by inhibiting the TLR4NFκB/NLRP3 signaling pathway.

Tanshinone IIA ameliorates myocardial ischemia/reperfusion injury in rats by regulation of NLRP3 inflammasome activation and Th17 cells differentiation

Purpose: Tanshinone IIA is a well-known lipophilic active constituent refined from traditional Chinese medicines, danshen. It has been previously demonstrated to possess various biological properties, including anti-inflammatory, antioxidant, promoting angiogenesis effect and so on. However, the mechanism of tanshinone IIA on myocardial ischemia-reperfusion injury (MI/RI) remains unclear. In this study, we investigated the effect of tanshinone IIA on MI/RI. Methods: MI/RI rat models were set up. Echocardiographic evaluation and hematoxylin and eosin staining were performed to analyze the cardiac function and morphology of MI/RI. Western blot was conducted for the detection of protein expression of pyrin domain containing 3 (NLRP3) and caspase-1 in heart tissues. Flow cytometry and real-time polymerase chain reaction were used for the detection of proinflammatory cytokines and Th17 cells differentiation. Results: We found that tanshinone IIA alleviated the myocardial damage of MI/RI, ameliorated the overall and local inflammatory reaction, and produced a cardioprotective effect by inhibiting of NLRP3 inflammasome activation and Th17/Treg cells differentiation. Conclusions: Our results highlighted the cardio-protective effect of tanshinone IIA on MI/RI and uncovered its underlying mechanism related to the NLRP3 inflammasome inhibition and the modulation of Th17/Treg cells differentiation.

Desfechos clínicos de pacientes após uso de terapia fibrinolítica pré-hospitalar: revisão sistemática / Resultados clínicos de pacientes después del uso de terapia fibrinolítica prehospitalaria: revisión sistemática / Clinical outcomes of patients after using prehospital fibrinolytic therapy: a systematic review

Resumo Objetivo: Comparar desfechos clínicos de óbito, reinfarto e Acidente Vascular Encefálico (AVE) em estudos primários que avaliaram o uso da Terapia Fibrinolítica (TF) em relação à Intervenção Coronariana Percutânea Primária (ICPP) para reperfusão miocárdica em pacientes com Infarto Agudo do Miocárdio com supradesnivelamento do segmento ST (IAMCST) no atendimento pré-hospitalar. Método: Revisão sistemática de literatura com busca realizada nas bases de dados CINAHL, MEDLINE, PUBMED, Science Direct, SCOPUS e Web of Science no período de outubro a dezembro de 2017. Foram incluídos Ensaios Clínicos Randomizados, disponíveis na íntegra, em qualquer idioma, sem recorte temporal. A avaliação da elegibilidade foi realizada em duas etapas e aplicada a Escala de Jadad para avaliação metodológica dos estudos encontrados. Resultados: Foram incluídos cinco Ensaios Clínicos Randomizados. A TF pré-hospitalar apresentou taxas de mortalidade em 30 dias após a intervenção semelhantes à ICPP, enquanto que em cinco anos foram encontrados valores menores para a TF. O tratamento instituído em um período menor que duas horas dos sintomas iniciais apresentou associação com a diminuição da mortalidade quando foi utilizada a TF. O reinfarto não-fatal, acidente vascular encefálico e a hemorragia intracraniana foram maiores quando utilizada a TF, enquanto que o choque cardiogênico apresentou menor frequência. Conclusão: A TF foi mais eficaz no tratamento pré-hospitalar para a redução dos óbitos após cinco anos, entretanto, o reinfarto e o AVE ocorreram de forma semelhante na amostra analisada. O fator tempo reduziu os desfechos clínicos, principalmente quando a terapia implementada ocorreu em até duas horas após a ocorrência do IAMCST. Assim, apesar das intervenções terem apresentado desfechos semelhantes, entretanto, a TF pode representar um tratamento viável em locais onde a ICPP não pode ser alcançada em tempo hábil.

Resumen Objetivo: Comparar resultados clínicos de fallecimiento, reinfarto y accidente vascular encefálico (AVE) en estudios primarios que analizaron el uso de la terapia fibrinolítica (TF) respecto a la intervención coronaria percutánea primaria (ICPP) para reperfusión miocárdica en pacientes con infarto agudo de miocardio con supradesnivel del segmento ST (IAMCST) en la atención prehospitalaria. Método: Revisión sistemática de literatura con búsqueda realizada en las bases de datos CINAHL, MEDLINE, PUBMED, Science Direct, SCOPUS y Web of Science en el período de octubre a diciembre de 2017. Se incluyeron ensayos clínicos aleatorizados, con texto completo disponible, en cualquier idioma, sin recorte temporal. El análisis de elegibilidad se realizó en dos etapas y se aplicó la escala de Jadad para una evaluación metodológica de los estudios encontrados. Resultados: Se incluyeron cinco ensayos clínicos aleatorizados. La TF prehospitalaria presentó índices de mortalidad 30 días después de la intervención semejantes a la ICPP, mientras que en cinco años se encontraron valores menores en la TF. El tratamiento aplicado en un período menor a dos horas desde los síntomas iniciales presentó una relación con la reducción de la mortalidad cuando se utilizó la TF. Los reinfartos no fatales, los accidentes vasculares encefálicos y las hemorragias intracerebrales fueron mayores cuando se utilizó la TF, mientras que los choques cardiogénicos presentaron menor frecuencia. Conclusión: La TF fue más eficaz en el tratamiento prehospitalario para reducir los fallecimientos después de cinco años, sin embargo, los reinfartos y los AVE ocurrieron de forma semejante en la muestra analizada. El factor tiempo redujo los resultados clínicos, principalmente cuando la terapia implementada ocurrió hasta dos horas después del episodio del IAMCST. De esta forma, a pesar de que las intervenciones presentaron resultados semejantes, la TF puede representar un tratamiento viable en lugares donde la ICPP no puede realizarse a tiempo.

Abstract Objective: To compare clinical outcomes of death, reinfarction, and stroke in primary studies assessing Fibrinolytic Therapy (FT) use in relation to Primary Percutaneous Coronary Intervention (PPCI) for myocardial reperfusion in patients with ST-Elevation Myocardial Infarction (STEMI) in prehospital care. Method: A systematic literature review conducted in the CINAHL, MEDLINE, PUBMED, Science Direct, SCOPUS, and Web of Science databases from October to December 2017. Randomized Clinical Trials, available in full, in any language, without temporal clipping were included. The eligibility assessment was carried out in two stages and applied to the Jadad Scale for methodological assessment of the studies found. Results: Five Randomized Clinical Trials were included. Prehospital FT presented mortality rates at 30 days after the intervention similar to PPCI, while in five years lower values were found for FT. The treatment instituted in a period of less than two hours of the initial symptoms was associated with the decrease in mortality when FT was used. Non-fatal reinfarction, stroke and intracranial hemorrhage were higher when FT was used, while cardiogenic shock showed lower frequency. Conclusion: FT was more effective in prehospital treatment to reduce deaths after five years, however, reinfarction and stroke occurred similarly in the sample analyzed. The time factor reduced clinical outcomes, especially when the implemented therapy occurred within two hours after the occurrence of STEMI. Thus, although the interventions presented similar outcomes. However, FT may represent a viable treatment in places where PPCI cannot be achieved in a timely manner.

Cardioprotective effects of pharmacological blockade of the mitochondrial calcium uniporter on myocardial ischemia-reperfusion injury

Abstract Purpose To evaluate whether the attenuation of mitochondrial Ca2+ overload produced by pharmacological blockade of mitochondrial Ca2+ uniporter (MCU) protects the myocardium against injuries caused by cardiac ischemia and reperfusion (CIR). Methods CIR was induced in adult male Wistar rats (300-350 g) by occlusion of the left anterior descendent coronary artery (10 min), followed by reperfusion (120 min). Rats were treated with different doses of MCU blocker ruthenium red (RuR), administered 5 min before ischemia or reperfusion. Results In untreated rats, the incidences of ventricular arrhythmias (VA), atrioventricular block (AVB) and the lethality (LET) induced by CIR were 85%, 79% and 70%, respectively. In rats treated with RuR before ischemia, the incidences of VA, AVB and LET were significantly reduced to 62%, 25% and 25%, respectively. In rats treated with RuR after ischemia, the incidences of VA, AVB and LET were significantly reduced to 50%, 25% and 25%, respectively. Conclusion The significant reduction of the incidence of CIR-induced VA, AVB and LET produced by the treatment with RuR indicates that the attenuation of mitochondrial Ca2+ overload produced by pharmacological blockade of MCU can protect the myocardium against injuries caused by CIR.

GDF11 ameliorated myocardial ischemia reperfusion injury by antioxidant stress and up-regulating autophagy in STZ-induced type 1 diabetic rats

Abstract Purpose: To investigate whether GDF11 ameliorates myocardial ischemia reperfusion (MIR) injury in diabetic rats and explore the underlying mechanisms. Methods: Diabetic and non-diabetic rats subjected to MIR (30 min of coronary artery occlusion followed by 120 min of reperfusion) with/without GDF11 pretreatment. Cardiac function, myocardial infarct size, creatine kinase-MB (CK-MB), lactate dehydrogenase (LDH), superoxide dismutase (SOD) 15-F2tisoprostane, autophagosome, LC3II/I ratio and Belcin-1 level were determined to reflect myocardial injury, oxidative stress and autophagy, respectively. In in vitro study, H9c2 cells cultured in high glucose (HG, 30mM) suffered hypoxia reoxygenation (HR) with/without GDF11, hydrogen peroxide (H2O2) and autophagy inhibitor 3-methyladenine (3-MA) treatment, cell injury; oxidative stress and autophagy were assessed. Results: Pretreatment with GDF11 significantly improved cardiac morphology and function in diabetes, concomitant with decreased arrhythmia severity, infarct size, CK-MB, LDH and 15-F2tisoprostane release, increased SOD activity and autophagy level. In addition, GDF11 notably reduced HR injury in H9c2 cells with HG exposure, accompanied by oxidative stress reduction and autophagy up-regulation. However, those effects were completely reversed by H2O2 and 3-MA. Conclusion: GDF11 can provide protection against MIR injury in diabetic rats, and is implicated in antioxidant stress and autophagy up-regulation.

Astragaloside IV alleviates myocardial ischemia-reperfusion injury in rats through regulating PI3K/AKT/GSK-3β signaling pathways

Abstract Purpose: To investigate the effect of astragaloside IV (As-IV) on myocardial ischemia-reperfusion (I/R) injury in rats and reltaed mechanisms. Methods: Sixty rats were randomly divided into sham-operated, control I/R and 2.5, 5 and 10 mg/kg As-IV groups, 12 rats in each group. The later three groups were intragastrically administered with As-IV for 7 days, with a dose of 2.5, 5 and 10 mg/kg, respectively. The myocardial I/R injury model was constructed in later four groups. At the end of reperfusion, the cardiac function indexes, serum lactate dehydrogenase (LDH) and creatine kinase (CK) levels, heart weight (HW)/body weight (BW) ratio and infarct size, and expressions of phosphatidylinositol-3 kinase/serine-threonine protein kinase (PI3K/AKT) and glycogen synthase kinase-3β (GSK-3β) proteins and the phosphorylated forms (p-AKT, p-GSK-3β) were determined. Results: Compared with control I/R group, in 5 and 10 mg/kg As-IV groups the left ventricular systolic pressure, fractional shortening and ejection fraction were increased, the left ventricular end-diastolic pressure was decreased, the serum LDH and CK levels were decreased, the HW/BW ratio and myocardial infarct size were decreased, and the p-Akt/Akt ratio and p-GSK-3β/GSK-3β ratio were increased (all P < 0.05). Conclusion: As-IV can alleviate the myocardial I/R injury in rats through regulating PI3K/AKT/GSK-3β signaling pathways.

Effect of miR-134 against myocardial hypoxia/reoxygenation injury by directly targeting NOS3 and regulating PI3K/Akt pathway

Abstract Purpose To reveal the function of miR-134 in myocardial ischemia. Methods Real-time PCR and western blotting were performed to measure the expression of miR-134, nitric oxide synthase 3 (NOS3) and apoptotic-associated proteins. Lactic dehydrogenase (LDH) assay, cell counting kit-8 (CCK-8), Hoechst 33342/PI double staining and flow cytometry assay were implemented in H9c2 cells, respectively. MiR-134 mimic/inhibitor was used to regulate miR-134 expression. Bioinformatic analysis and luciferase reporter assay were utilized to identify the interrelation between miR-134 and NOS3. Rescue experiments exhibited the role of NOS3. The involvement of PI3K/AKT was assessed by western blot analysis. Results MiR-134 was high regulated in the myocardial ischemia model, and miR-134 mimic/inhibitor transfection accelerated/impaired the speed of cell apoptosis and attenuated/exerted the cell proliferative prosperity induced by H/R regulating active status of PI3K/AKT signaling. LDH activity was also changed due to the different treatments. Moreover, miR-134 could target NOS3 directly and simultaneously attenuated the expression of NOS3. Co-transfection miR-134 inhibitor and pcDNA3.1-NOS3 highlighted the inhibitory effects of miR-134 on myocardial H/R injury. Conclusion This present work puts insights into the crucial effects of the miR-134/NOS3 axis in myocardial H/R injury, delivering a potential therapeutic technology in future.

Pharmacological modulation of b-adrenoceptors as a new cardioprotective strategy for therapy of myocardial dysfunction induced by ischemia and reperfusion

Abstract Purpose: To evaluate the cardioprotective response of the pharmacological modulation of β-adrenergic receptors (β-AR) in animal model of cardiac ischemia and reperfusion (CIR), in spontaneously hypertensive (SHR) and normotensive (NWR) rats. Methods: CIR was induced by the occlusion of left anterior descendent coronary artery (10 min) and reperfusion (75 min). The SHR was treated with β-AR antagonist atenolol (AT, 10 mg/kg, IV) 5 min before CIR, and NWR were treated with β-AR agonist isoproterenol (ISO, 0.5 mg/kg, IV) 5 min before CIR. Results: The treatment with AT increased the incidence of VA, AVB and LET in SHR, suggesting that spontaneous cardioprotection in hypertensive animals was abolished by blockade of β-AR. In contrast, the treatment with ISO significantly reduced the incidence of ventricular arrhythmia, atrioventricular blockade and lethality in NWR (30%, 20% and 20%, respectively), suggesting that the activation of β-AR stimulate cardioprotection in normotensive animals. Serum CK-MB were higher in SHR/CIR and NWR/CIR compared to respective SHAM group (not altered by treatment with AT or ISO). Conclusion: The pharmacological modulation of β-AR could be a new cardioprotective strategy for the therapy of myocardial dysfunctions induced by CIR related to cardiac surgery and cardiovascular diseases.

Tramadol Alleviates Myocardial Injury Induced by Acute Hindlimb Ischemia Reperfusion in Rats / Tramadol Atenua Lesões Miocárdicas Induzidas pela Reperfusão Pós- Isquêmica Aguda de Membros Posteriores em Ratos

AbstractBackground:Organ injury occurs not only during periods of ischemia but also during reperfusion. It is known that ischemia reperfusion (IR) causes both remote organ and local injuries.Objective:This study evaluated the effects of tramadol on the heart as a remote organ after acute hindlimb IR.Methods:Thirty healthy mature male Wistar rats were allocated randomly into three groups: Group I (sham), Group II (IR), and Group III (IR + tramadol). Ischemia was induced in anesthetized rats by left femoral artery clamping for 3 h, followed by 3 h of reperfusion. Tramadol (20 mg/kg, intravenous) was administered immediately prior to reperfusion. At the end of the reperfusion, animals were euthanized, and hearts were harvested for histological and biochemical examination.Results:The levels of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) were higher in Groups I and III than those in Group II (p < 0.05). In comparison with other groups, tissue malondialdehyde (MDA) levels in Group II were significantly increased (p < 0.05), and this increase was prevented by tramadol. Histopathological changes, including microscopic bleeding, edema, neutrophil infiltration, and necrosis, were scored. The total injuryscore in Group III was significantly decreased (p < 0.05) compared with Group II.Conclusion:From the histological and biochemical perspectives, treatment with tramadol alleviated the myocardial injuries induced by skeletal muscle IR in this experimental model.

ResumoFundamento:Lesões a órgãos ocorrem não apenas durante períodos de isquemia, mas paradoxalmente, também durante a reperfusão. Sabe-se que a reperfusão pós-isquêmica (RPI) causa lesões tanto remotas quanto locais no órgão afetado.Objetivo:Este estudo avaliou os efeitos do tramadol no coração como órgão remoto, após RPI aguda dos membros posteriores.Métodos:Trinta ratos Wistar, machos, adultos e saudáveis, foram distribuídos aleatoriamente em três grupos: Grupo I (controle), Grupo II (RPI) e Grupo III (RPI + tramadol). Isquemia foi induzida em ratos anestesiados através do pinçamento da artéria femoral esquerda por 3 horas, seguidas de 3 horas de reperfusão. Tramadol foi administrado (20 mg/kg, IV) imediatamente antes da reperfusão. Ao final da reperfusão, os animais foram sacrificados e seus corações coletados para exames histológicos e bioquímicos.Resultados:Os níveis de superóxido-dismutase (SOD), catalase (CAT) e glutationa-peroxidase (GPx) foram maiores nos grupos I e III que no grupo II (p < 0.05). Em comparação aos outros grupos, os níveis tissulares de malondialdeído (MDA) estavam significativamente mais elevados no grupo II (p < 0.05), o que foi evitado pelo uso de tramadol. Foram pontuadas as alterações histopatológicas, incluindo micro-hemorragia, edema, infiltração por neutrófilos e necrose. A pontuação total das lesões do grupo III foi significativamente menor (p < 0.05) em comparação ao grupo II.Conclusão:Do ponto de vista histológico e bioquímico, o tratamento com tramadol diminuiu as lesões miocárdicas induzidas pela RPI da musculatura esquelética neste modelo experimental.

Inhibition of Janus activated kinase-3 protects against myocardial ischemia and reperfusion injury in mice

Recent studies have documented that Janus-activated kinase (JAK)-signal transducer and activator of transcription (STAT) pathway can modulate the apoptotic program in a myocardial ischemia/reperfusion (I/R) model. To date, however, limited studies have examined the role of JAK3 on myocardial I/R injury. Here, we investigated the potential effects of pharmacological JAK3 inhibition with JANEX-1 in a myocardial I/R model. Mice were subjected to 45 min of ischemia followed by varying periods of reperfusion. JANEX-1 was injected 1 h before ischemia by intraperitoneal injection. Treatment with JANEX-1 significantly decreased plasma creatine kinase and lactate dehydrogenase activities, reduced infarct size, reversed I/R-induced functional deterioration of the myocardium and reduced myocardial apoptosis. Histological analysis revealed an increase in neutrophil and macrophage infiltration within the infarcted area, which was markedly reduced by JANEX-1 treatment. In parallel, in in vitro studies where neutrophils and macrophages were treated with JANEX-1 or isolated from JAK3 knockout mice, there was an impairment in the migration potential toward interleukin-8 (IL-8) and monocyte chemoattractant protein-1 (MCP-1), respectively. Of note, however, JANEX-1 did not affect the expression of IL-8 and MCP-1 in the myocardium. The pharmacological inhibition of JAK3 might represent an effective approach to reduce inflammation-mediated apoptotic damage initiated by myocardial I/R injury.

CPU0213, a novel endothelin type A and type B receptor antagonist, protects against myocardial ischemia/reperfusion injury in rats

The efficacy of endothelin receptor antagonists in protecting against myocardial ischemia/reperfusion (I/R) injury is controversial, and the mechanisms remain unclear. The aim of this study was to investigate the effects of CPU0123, a novel endothelin type A and type B receptor antagonist, on myocardial I/R injury and to explore the mechanisms involved. Male Sprague-Dawley rats weighing 200-250 g were randomized to three groups (6-7 per group): group 1, Sham; group 2, I/R + vehicle. Rats were subjected to in vivo myocardial I/R injury by ligation of the left anterior descending coronary artery and 0.5 percent sodium carboxymethyl cellulose (1 mL/kg) was injected intraperitoneally immediately prior to coronary occlusion. Group 3, I/R + CPU0213. Rats were subjected to identical surgical procedures and CPU0213 (30 mg/kg) was injected intraperitoneally immediately prior to coronary occlusion. Infarct size, cardiac function and biochemical changes were measured. CPU0213 pretreatment reduced infarct size as a percentage of the ischemic area by 44.5 percent (I/R + vehicle: 61.3 ± 3.2 vs I/R + CPU0213: 34.0 ± 5.5 percent, P < 0.05) and improved ejection fraction by 17.2 percent (I/R + vehicle: 58.4 ± 2.8 vs I/R + CPU0213: 68.5 ± 2.2 percent, P < 0.05) compared to vehicle-treated animals. This protection was associated with inhibition of myocardial inflammation and oxidative stress. Moreover, reduction in Akt (protein kinase B) and endothelial nitric oxide synthase (eNOS) phosphorylation induced by myocardial I/R injury was limited by CPU0213 (P < 0.05). These data suggest that CPU0123, a non-selective antagonist, has protective effects against myocardial I/R injury in rats, which may be related to the Akt/eNOS pathway.

Trimetazidina na injúria de isquemia e reperfusão em cirurgia de revascularização do miocárdio / Trimetazidine on ischemic injury and reperfusion in coronary artery bypass grafting

FUNDAMENTO: A injúria de isquemia e reperfusão constitui um mecanismo fisiopatológico frequente e de difícil controle durante a Cirurgia de Revascularização do Miocárdio (CRVM) com circulação extracorpórea, sendo o momento crítico o término da cirurgia, quando ocorre o desclampeamento da aorta e a liberação dos radicais hiperóxidos causadores da injúria. OBJETIVO: Avaliar, em estudo prospectivo, duplo-cego randomizado, controlado com placebo, os efeitos da Trimetazidina (Tmz) sobre a injúria de isquemia e reperfusão miocárdica, identificando a variação dos marcadores plasmáticos de agressão miocárdica (troponina T e Cpk-Mb), e as alterações ecocardiográficas da função ventricular. MÉTODOS: Foram estudados 60 pacientes, divididos em dois grupos (Placebo e Tmz) com, no máximo, disfunção ventricular leve, estratificados por ecocardiografia e recebendo medicação/placebo na dose - no pré-operatório sem medicação, 12 a 15 dias de medicação/placebo colhida cinco minutos após o desclampeamento aórtico, e nas 12, 24 e 48 horas seguintes. RESULTADOS: Tanto a troponina T como a CpK-Mb atingiram valores altamente significativos (p = 0,0001) no grupo tratado em relação ao grupo controle nos quatro momentos analisados − 5 min, 12 h, 24 h e 48 h. As variáveis ecocardiográficas não evidenciaram mudanças evolutivas em cada grupo isoladamente e quando comparados em conjunto. CONCLUSÃO: A trimetazidina mostrou-se eficaz na redução da injúria de isquemia e reperfusão, não interferiu na função ventricular esquerda, e não foram observados efeitos colaterais.

BACKGROUND: The ischemia and reperfusion ischemia is a common physiopathological mechanisms, which has difficult control during Coronary Artery Bypass Grafting (CABG) with cardiopulmonary bypass, the critical moment of which happening by the end of surgery, when there is declamping of aorta and release of hyperoxic radicals causing the injury. OBJECTIVE: Evaluate, in a randomized double-blind prospective study, controlled with placebo, the effects of Trimetazidine (Tmz) on ischemic injury and myocardial reperfusion, identifying the change in plasma markers of a myocardial aggression (troponin T and CPK-MB), and echocardiographic changes of ventricular function. METHODS: We studied 60 patients divided in two groups (placebo and Tmz) with mild ventricular dysfunction at the most, stratified by echocardiography and receiving medication/placebo at a dose of 20 mg/3x/day, starting from 12 to 15 days after pre-operative period up to 5 to 8 days after post-operative period. Troponin T and Cpk-Mb were measured preoperatively without medication, 12 to 15 days of medication/placebo taken five minutes after aortic declamping, and at subsequent 12, 24 and 48 hours. RESULTS: Both Troponin T and Cpk-Mb reached highly significant values (p = 0.0001) in the treated group compared to the control group at the four moments analyzed - 5 min, 12h, 24h and 48h. The echocardiographic variables did not show evolutive changes in each group severally considered and when compared among themselves. CONCLUSION: Trimetazidine was effective in reducing ischemic injury and reperfusion, had no effect on left ventricular function, and no side effects were observed.

Proteção da recuperação funcional do miocárdio pelo omeprazol após isquemia-reperfusão em corações isolados de ratos / Myocardium functional recovery protection by omeprazole after ischemia-reperfusion in isolated rat hearts

OBJETIVO: Analisar efeitos do omeprazol na proteção da recuperação funcional de corações isolados de ratos submetidos à lesão de isquemia-reperfusão. MÉTODOS: Foram estudados 12 ratos Wistar, peso corpóreo médio de 280g. Após anestesia com injeção intra-abdominal de 10mg de cetamina e 2mg de xilazina, os corações foram removidos e mantidos em perfusão com solução Krebs-Henseleit (95 por centoO2 e 5 por cento CO2, 37ºC, 110-120mmHg de pressão de perfusão e pressão diastólica de 8 mmHg) em sistema Langendorff, modificado, descartável, modelo FCSFA-ServCor (Comex Ltda.). Os seis corações do Grupo I (GI) e os seis do Grupo II (GII) foram submetidos a 20 minutos de isquemia e 30 minutos de reperfusão. Nos corações do Grupo II, imediatamente antes da isquemia, foram administrados via perfusão coronária 200mcg de omeprazol. Foram controlados frequência cardíaca (FC), fluxo coronário (FCo), pressão sistólica (PS), +dP/dt e -dP/dt, após estabilização (t0) e no final da reperfusão (t30). Empregou-se método não paramétrico de Kruskal-Wallis (P<0,05) para análise estatística dos dados. RESULTADOS: Não ocorreram diferenças (P>0,05) entre os valores de FCo e FC nos dois grupos. No final do período de reperfusão (t30), foram significantes (P<0,05) as variações da PS reduzida para 28,0±3,6 por cento do valor inicial (t0) no Grupo I e para 79.0±5,9 por cento no Grupo II; a +dP/dtmax declinou para 31,0±5,6 por cento no GI, mantendo-se em 99,4±11,2 por cento (P<0,05) no GII e a -dP/dtmax declinou para 26,0±7,3 por cento no GI, mantendo-se em 82,0±2, 2 por cento no GII (P<0,05). CONCLUSÃO: A administração do omeprazol antes da indução da isquemia coronária protegeu significantemente a recuperação funcional do miocárdio.

OBJECTIVE: To evaluate the myocardium contractility alterations of isolated hearts of rats, submitted to ischemia and reperfusion with and without administration of the omeprazole. METHODS: Twelve Wistar breed rats with 270g mean body weight was studied. After anesthesia by intraperitoneal injection of ketamine 10mg and xylazine 2mg, their hearts were removed and perfused with Krebs-Henseleit solution (95 percent of O2 and 5 percent of CO2, 37ºC, 110-120 mmHg perfusion pressure, 8 mmHg ventricular diastolic pressure) in the São Francisco de Assis disposable Langendorff system model Comex Ltda, MG. The six hearts of Group I (GI) and of the Group II (GII) were submitted to 20 min ischemia and 30 min reperfusion. In GII hearts, intracoronary injection of omeprazole 200 mcg was done immediately before the ischemia period induction. The following parameters were registered after the stabilization period (t0), and after the reperfusion period (t30): heart rate (HR), coronary flow (CoF), systolic pressure (SP), +dP/dt and -dP/dt. The Kruskal-Wallis test (P<0.05) was applied to statistical analysis. RESULTS: There were no significant differences (P>0.05) between groups among HR and CoF values. Differences (P<0.05) occurred between groups, I e II after the reperfusion period (t30) regarding systolic pressure reduced for 28.0±3.6 percent in the control group GI and for 79.0±5.9 percent in GII; The +dP/dtmax declined to be only 31.0±5.6 percent in GI, preserving 99.4±11.2 percent values in GII (P<0.05). The t30 -dP/dtmax values were GI 26.0±7.3 percent and GII 82.0±2.2 percent (P<0.05). CONCLUSION: The omeprazole administration before ischemia induction significantly protected the myocardium function recovery.

effect of aminophylline versus milrinone on ischemia-reperfusion myocardial injury during open heart surgery

Cardioplegic arrest during cardiopulmonary bypass [CPB] is essential for the majority of cardiac surgical procedures; Cardioplegia protects the myocardium by providing continuous or intermittent oxygen while simultaneously reducing cardiomyocyte oxygen demand, but it does not inherently increase the ischemic-reperfusion injury tolerance of the cardiomyocytes. Aminophylline and milrinone by their phosphodiesterase inhibitor and anti-inflammatory activity may decrease this type of injury. This study has been designed to compare between the protective effect of aminophylline and milrinone during open heart surgery for valve replacement with CPB. Sixty adult patients undergoing elective single valve replacement were randomized to receive aminophylline 5 mg/kg [n=20], milrinone 50 -microg/kg [n=20], or normal saline as control group [n=20] through intravenous infusion 10 minutes before the aortic cross-clamping. The cardiac troponin I, inotrope score, duration of mechanical ventilation, and length of ICU stay and other hemodynamic variables were measured and recorded. There were no differences between the three groups with regard to clinical variables. Cardiac troponin I raised significantly after declamping in the three groups, however it was significantly lower in aminophylline and milrinone group compared to control group immediately after CBP and after 8 hours with no significant differences between aminophylline and milrinone group, inotrope score duration of mechanical ventilation and length of ICU stay showed no significant differences between the three groups. Administration of aminophylline or milrinone reduces the subclinical myocardial injury with no difference between both agents and with no effect on the hemodynamic parameters or short term clinical outcome in patients undergoing single valve replacement with CPB

effect of repeated diazepam administration on myocardial function in the ischemia-reperfused isolated rat heart

To evaluate whether repeated diazepam administration affects the heart in ischemia- reperfusion. This study was performed at the Medical Biology Research Center, Kermanshah, Iran, from March to September 2008. Four groups of rats were subjected to a daily injection of diazepam [group 1 [0.5 mg/kg for 21 days], group II [2.5 mg/kg for 5 days], and group III [5 mg/kg for 5 days] intraperitoneally], and saline solution [21 days] in the control groups. Isolated, perfused hearts were subjected to 40 minutes global ischemia, and 45 minutes reperfusion. The left ventricular developed pressure [LVDP], heart rate, and coronary flow were measured. Rate pressure product [RPP] was calculated. In reperfusion, released lactate dehydrogenase [LDH] enzyme in effluent was measured. It was observed that the recovery of the RPP and LVDP in reperfusion significantly decreased in the test group III [n=9] in comparison to the control [n=8]. During the reperfusion period, the released LDH significantly increased in test group II [n=8] and group III in comparison with the control. The results show that repeated administration of diazepam [5 mg/kg for 5 days] reduced the cardiac performance in reperfusion, and significantly exacerbated the ischemia-reperfusion injury. It is probably mediated by the changing of cardiac susceptibility in ischemia due to repeated administration of diazepam

The Effect of High Dose Melatonin on Cardiac Ischemia-reperfusion Injury

PURPOSE: Melatonin, the most potent scavenger of toxic free radicals, has been found to be effective in protecting against pathological states due to the release of reactive oxygen species. This study was performed to establish the effect of high dose melatonin on protection against ischemia- reperfusion (I/R) injury in rat hearts. MATERIALS AND METHODS: Forty male Sprague-Dawley rats were used in this study. They were separated into four groups of ten rats each. A left coronary artery occlusion was induced in the rats by ligating the artery for 20 minutes and then releasing the ligation (reperfusion) afterwards. The control group was Group A. Group B was subjected to myocardial ischemia-reperfusion without any treatment, while Group C underwent myocardial ischemia-reperfusion with a melatonin treatment before the ischemia. Group D was subjected to myocardial ischemia-reperfusion with a melatonin treatment before the reperfusion. After 20 minutes of reperfusion, blood samples were obtained from each group for biochemical studies, and the animals were sacrificed for histological and, immunohistochemical examinations of the myocardial tissue. RESULTS: We found that the cardiac troponin T(cTn-T) levels were significantly increased in Group B when all groups were compared. In the Group C rats treated with melatonin, the cTn-T values were significantly lower than those in Groups B and D. In addition, malondialdehyde (MDA) and antioxidant enzymes including, superoxide dismutase (SOD) and myeloperoxidase (MPO) were lower than those in Group B in the melatonin treated groups. The differences were statistically significant (p < 0.05). Histopathologic and immunohistopathologic studies also supported the effectiveness of melatonin. CONCLUSION: Our study suggests that high dose melatonin, appears to offer protection against cardiac ischemia-reperfusion injuries in rats by scavenging the free radicals and could have a potential clinical use in the management of myocardial ischemia.

Utilidad de la trimetazidina en la lesión por isquemia-reperfusión: Estudio experimental en un modelo canino de aturdimiento miocárdico / Usefulness of trimetazidine in ischemia-reperfusion lesion: experimental study in myocardial stunning model

Objetivo: Evaluar la acción de la trimetazidina en el deterioro de la función sistólica que se produce en el miocardio tras una isquemia única y prolongada. Métodos: Se analizaron 13 perros mestizos, de uno u otro sexo, asignados al azar a tratamiento oral con trimetazidina (6 perros) o placebo (7 perros) durante 7 días. Se realizó un protocolo de isquemia bajo anestesia consistente en una obstrucción completa de la arteria coronaria descendente anterior de 15 minutos de duración, seguida de 60 minutos de reperfusión. Las variables analizadas durante la obstrucción y la reperfusión fueron: Frecuencia cardíaca (FC), presión ventricular izquierda (PVI), dP/dt y las curvas de función regional de la zona isquémica y de una zona testigo (longitud telediastólica, telesistólica y fracción de acortamiento). Resultados: Las variables hemodinámicas (FC, PVI y dP/dt), no presentaron diferencias significativas entre ambos grupos, con poca variabilidad de sus valores respecto a los basales durante la isquemia-reperfusión. La fracción de acortamiento de la zona isquémica experimentó una disminución estadísticamente significativa durante la obstrucción coronaria en ambas series, alcanzando valores de discinesia, con persistencia de la disfunción contráctil tras 60 minutos de reperfusión, y sin diferencias entre ambas series (50% serie Placebo; 41% serie Trimetazidina). Conclusiones: La recuperación de la contractilidad miocárdica tras una isquemia completa en la serie tratada con TMZ no mostró diferencias significativas respecto a la serie Placebo, a diferencia de lo que ocurre con períodos de oclusión más cortos y repetidos.

Objective: The aim of this study is to evaluate the effect of trimetazidine (TMZ) on myocardial systolic dysfunction resulting from an isolated episode of induced coronary ischemia. Methods: In a double-blinded randomized design we studied 13 mongrel anesthetized dogs of either sex (6 of them treated with oral TMZ previously). The anterior descending coronary artery was totally occluded during 15 minutes followed by 60 minutes of reperfusion. Global and regional cardiac variables were recorded in control and ischemic areas. Results: There were no significant differences between TMZ and placebo series with respect to global cardiac function variables. Both series showed no significant variations in global variables during the ischemia-reperfusion process. The shortening fraction in the ischemic area fell significantly during the ischemic period in both TMZ and placebo series reaching dyskinetic values. Myocardial contractility dysfunction persisted after 60 minutes of reperfusion in both series with no significant differences (41% vs 50% placebo). Conclusions: Contrary to shorter and repeated occlusion periods, myocardial contractility recovery after a complete episode of ischemia did not show significant differences between TMZ-treated and placebo series.

Solución GIK en el infarto agudo al miocardio: el resurgimiento de una antigua terapia / Glucose-insulin-K solution for acute myocardial infarction: rebirth of an old form of therapy

La terapia con solución de glucosa insulina y potasio en el infarto o solución GIK fue inicialmente utilizada por Sodi-Pallares. Desde entonces muchos trabajos con esta solución han sido publicados con resultados disímiles. Sin embargo el resultado de un meta-análisis reciente, que incluye sólo trabajos randomizados con dosis adecuadas de GIK, parece confirmar la disminución de la mortalidad asociada a solución GIK. Para comprender mejor los fundamentos y posibles mecanismos de beneficio con el empleo de la solución GIK en el infarto del miocardio, revisaremos primero el metabolismo miocárdico normal y en condiciones de isquemia, luego el daño por reperfusión post infarto y los efectos de la solución GIK en el miocardio. Por último, analizaremos las experiencias clínicas publicadas con esta terapia