Myocardial Ischemia-reperfusion Damage Research Articles

Activating Hypoxia-Inducible Factor-1α Reduces Myocardial Ischemia-Reperfusion Injury in Mice Through Hexokinase II

To verify that HIF-1α/HKII pathway is the key mechanism to alleviate myocardial ischemiareperfusion (IR) injury in aged mice through HIF-1α and HKII inhibitors. A mouse I/R model was carried out in young and old C57BL/6 mice for 60 min and reperfusion for 120 min. Mice were injected intraperitoneally with AAV-9 virus to introduce HIF-1α 24 h before ischemia. After 2 h of reperfusion, the mitochondrial ultrastructure, ATP content, membrane potential, and protein expression of HIF-1α, LC3, Bax, Bcl-2, Caspese-9, Caspase-3, and Cyt-3 were detected. After 24 h, the myocardial infarction area and cardiac ability were evaluated. Young mice and old mice have different protective effects after acute ischemia/reperfusion injury. After the introduction of HIF-1α by AAV-9 virus, the expression of the downstream target gene HKII can be up-regulated. At the same time, it reduces the expression of key proteins LC3, Bax, Caspese-9, and Caspase-3. Stabilize the membrane potential, ultimately reduce the area of myocardial infarction and improve heart function. Young mice and old mice have different protective effects after acute ischemia/reperfusion injury. The I/R in the older is caused by HIF-1α. HIF-1α inhibits the opening of mPTP by up-regulating HKII, stabilizes mitochondrial membrane potential, protects mitochondrial integrity, and reduces myocardial ischemia-reperfusion damage to the myocardium of elderly mice.

Phenols and terpenoids: natural products as inhibitors of NLRP3 inflammasome in cardiovascular diseases.

Inflammatory infiltration has been implicated in the pathogenesis of cardiovascular diseases (CVDs). The NLRP3 inflammasome is involved in the development of several types of CVDs, including myocardial infarction, myocardial ischemia-reperfusion damage, heart failure, atrial fibrillation, and hypertension. Inhibiting the activity of NLRP3 inflammasome can inhibit the progress of CVDs. However, there is no NLRP3 inflammasome inhibitor in clinic, and it is very important to find a safe and effective NLRP3 inhibitor. Phenols and terpenoids are naturally natural products that have many anti-inflammatory effects in CVDs by modulating the NLRP3 inflammatory pathway. Thus, 20 natural products from phenols and terpenoids for the treatment of cardiovascular disease based on the inhibition of NLRP3 inflammasome were summarized and screened. Docking results showed salvianolic acid B and ellagic acid in phenols, and oridonin and triptolide in terpenoids had a better binding activity with NLRP3, which can provide theoretical support for finding novel NLRP3 inflammasome inhibitors or lead compounds in the future.

Exosomal miR-455-3p from BMMSCs prevents cardiac ischemia-reperfusion injury

Bone marrow mesenchymal stem cells (BMMSCs) exert protective effects against myocardial infarction (MI). Here, we focused on the function and mechanism of miR-455-3p from BMMSCs-derived exosomes (BMMSCs-Exo) in myocardial infarction. BMMSCs were isolated from rat bone marrow, and the exosomes from the culture medium of BMMSCs were separated, and administered to H9C2 cells under hypoxia-reperfusion (H/R) stimulation. MTT and TUNEL staining analyzed cell viability and apoptosis, respectively. RT-qPCR determined miR-455-3p expression. Apoptosis-related proteins, autophagy-associated proteins, and the MEKK1-MKK4-JNK signaling pathway were detected. The interaction between miR-455-3p and MEKK1 was confirmed through dual luciferase activity and RIP assay. An in vivo ischemia reperfusion (I/R) model was established in rats. 2, 3, 5 triphenyltetrazolium chloride (TTC) staining, hematoxylin-eosin (H&E) staining, Masson staining, and TUNEL staining evaluated the infarct volume and histopathological changes. miR-455-3p's expression was down-regulated in BMMSCs-derived exosomes, I/R myocardial tissues, and H/R myocardial cells. miR-455-3p enriched by BMMSC exosomes reduced H/R-mediated cardiomyocyte damage and death-related autophagy. miR-455-3p upregulation suppressed MEKK1-MKK4-JNK. MEKK1 overexpression notably mitigated cell apoptosis, cramped cell viability, suppressed autophagy expansion, and attenuated Exo-miR-455-3p's protection on H/R myocardial cells. In-vivo trials reflected that BMMSC exosomes enriched with miR-455-3p repressed ischemia reperfusion-induced myocardial damage and myocardial cell function. miR-455-3p, shuttled by exosomes from MSCs, targets the MEKK1-MKK4-JNK signaling pathway to guard against myocardial ischemia-reperfusion damage.

LncRNA PVT1 Knockdown Ameliorates Myocardial Ischemia Reperfusion Damage via Suppressing Gasdermin D-Mediated Pyroptosis in Cardiomyocytes.

Objective: Myocardial ischemia reperfusion (I/R) damage is a life-threatening vascular emergency after myocardial infarction. Here, we observed the cardioprotective effect of long non-coding RNA (lncRNA) PVT1 knockdown against myocardial I/R damage.Methods: This study constructed a myocardial I/R-induced mouse model and a hypoxia/reoxygenation (H/R)-treated H9C2 cells. PVT1 expression was examined via RT-qPCR. After silencing PVT1 via shRNA against PVT1, H&E, and Masson staining was performed to observe myocardial I/R damage. Indicators of myocardial injury including cTnI, LDH, BNP, and CK-MB were examined by ELISA. Inflammatory factors (TNF-α, IL-1β, and IL-6), Gasdermin D (GSDMD), and Caspase1 were detected via RT-qPCR, western blot, immunohistochemistry, or immunofluorescence. Furthermore, CCK-8 and flow cytometry were presented for detecting cell viability and apoptosis.Results: LncRNA PVT1 was markedly up-regulated in myocardial I/R tissue specimens as well as H/R-induced H9C2 cells. Silencing PVT1 significantly lowered serum levels of cTnI, LDH, BNP, and CK-MB in myocardial I/R mice. H&E and Masson staining showed that silencing PVT1 alleviated myocardial I/R injury. PVT1 knockdown significantly lowered the production and release of inflammatory factors as well as inhibited the expression of GSDMD-N and Caspase1 in myocardial I/R tissue specimens as well as H/R-induced H9C2 cells. Moreover, silencing PVT1 facilitated cell viability and induced apoptosis of H/R-treated H9C2 cells.Conclusion: Our findings demonstrated that silencing PVT1 could alleviate myocardial I/R damage through suppressing GSDMD-mediated pyroptosis in vivo and in vitro. Thus, PVT1 knockdown may offer an alternative therapeutic strategy against myocardial I/R damage.

Rosuvastatin protects isolated hearts against ischemia-reperfusion injury: role of Akt-GSK-3β, metabolic environment, and mitochondrial permeability transition pore

The cardioprotective activity of rosuvastatin (R) is yet to be known. The objective of this study was to research whether R perfusion before global ischemia can mitigate myocardial ischemia-reperfusion damage, considering the metabolic condition in which these effects occur, and to contemplate potential mitochondrial benefits. Protein kinase B (Akt)/glycogen synthase kinase-3β (GSK-3β) and mitochondrial permeability transition pore (MPTP) are key elements in myocardial injury produced by ischemia-reperfusion. Isolated rat hearts were subjected to 25-min ischemia and 1-h reperfusion in the presence or absence of R, with or without Wortmannin (W), a phosphatidylinositol 3-kinase (PI3K)/Akt inhibitor. Akt and GSK-3β were measured by Western blot analysis; lactate, glycogen, and G6PDH were determined; and Ca2+-induced MPTP opening was evaluated using a spectrophotometric method. Contractility was assessed by left ventricular developed pressure (LVDP), and rate-pressure product (RPP), peak rate of contraction and peak rate of relaxation (± dP/dt), and left ventricular end-diastolic pressure (LVEDP) were determined. Tissue samples were extracted to evaluate mitochondrial damage by electron microscopy and to assess infarct size. Statistical analysis employed ANOVA (n = 6/per group). Myocardial infarct size was significantly reduced by R, which also improved cardiac function. MPTP opening was delayed to 300μM CaCl2, while use of W resulted in MPTP opening at 200μM CaCl2. Electron microscopy showed better mitochondrial preservation with R, which reduced lactic acid production, increased glycogen consumption and G6PDH activity, as well as phosphorylation of Akt and GSK-3β. R before ischemia is cardioprotective against ischemic and reperfusion damage, activating Akt and regulating GSK-3β negatively and attenuating the MPTP opening.

HIBISCUS-STEMI Biobank: High quality samples for powerful research on STEMI biomarkers

The HIBISCUS-STEMI BIOBANK (coHort of patients to Identify Biological and Imaging markerS of CardiovascUlar outcomes in ST elevation myocardial infarction) is based on a prospective cohort of 500 patients with ST elevation myocardial infarction (STEMI), supported by the RHU MARVELOUS (ANR-16-RHUS-0009). Its primary objective is to identify new biomarkers of clinical outcomes with specific attention to heart failure. All patients with suspicion of STEMI have given written consent and clinical, laboratory and imaging information has been recorded in the database of the Lyon Clinical Investigation Center (CIC). Blood sampling (taken 24/7) (24 mL) was repeated 5 times: before PCI (Percutaneous Coronary Intervention), at 4-24-48 hours and 1 month after PCI. Blood was immediately transferred to the local biobank to be processed in serum, plasma or peripheral blood mononuclear cells (PBMC), and cryopreserved at −80 °C or in liquid nitrogen. Total blood was also stored for future genetic studies. The biobank has been certified for NF S96-900 French norm and assure traceability and quality of samples and storage. More than 36,000 high quality biological samples associated with clinical and MRI (magnetic resonance imaging) data are already available and several projects are ongoing with new biomarkers being identified, e.g. to explore the role of inflammation in myocardial ischemia-reperfusion damage and development of LV remodeling and heart failure within 1 year after AMI. The HIBISCUS-STEMI Biobank is a powerful tool for biomarkers research, available for academic or industrial collaboration. This will help to better understand the mechanisms of AMI and, to improve our assessment of individual prognosis after acute myocardial infarction.

Sulforaphane protects from myocardial ischemia-reperfusion damage through the balanced activation of Nrf2/AhR

The activation of the transcription factor Nrf2 and the consequent increment in the antioxidant response might be a powerful strategy to contend against reperfusion damage. In this study we compared the effectiveness between sulforaphane (SFN), a well known activator of Nrf2 and the mechanical maneuver of post-conditioning (PostC) to confer cardioprotection in an in vivo cardiac ischemia-reperfusion model. We also evaluated if additional mechanisms, besides Nrf2 activation contribute to cardioprotection. Our results showed that SFN exerts an enhanced protective response as compared to PostC. Bot, strategies preserved cardiac function, decreased infarct size, oxidative stress and inflammation, through common protective pathways; however, the aryl hydrocarbon receptor (AhR) also participated in the protection conferred by SFN. Our data suggest that SFN-mediated cardioprotection involves transient Nrf2 activation, followed by phase I enzymes upregulation at the end of reperfusion, as a long-term protection mechanism.

Research progress on the effect of anesthesia on anthracycline cardiotoxicity

Anthracycline-based antitumor drugs are one of the highly effective broad-spectrum anti-cancer drugs that are commonly used clinically. However, anthracyclines have the serious side effect of cumulative dose-related cardiotoxicity. Clinically, intake of myocardial protective drugs is one of the most important strategies for the safe use of anthracyclines in chemotherapy. Numerous cancer patients un- dergoing chemotherapy also require surgery, during which general anesthesia is commonly used. Various studies have shown that propofol and sevoflurane, the most commonly used general anesthetics, present a significant protective effect in myocardial ischemia-reperfusion damage. Therefore, for patients who have just undergone chemotherapy with anthracyclines, anthracycline-induced myocardial injury may occur at the acute or chronic phase of surgery. Consequently, if the effect of anesthetics on the cardiotoxicity of anthracyclines is similar to the protective effect of these drugs for myocardial ischemia-reperfusion damage, this protective effect on cardiotoxicity will have significant impact on the recovery of cardiopulmonary function of patients after surgery. Thus, further studies are needed to establish the effect of anesthetics on the cardiotoxicity of anthracyclines.

Vasonatrin peptide attenuates myocardial ischemia-reperfusion injury in diabetic rats and underlying mechanisms.

Diabetes mellitus increases morbidity/mortality of ischemic heart disease. Although atrial natriuretic peptide and C-type natriuretic peptide reduce the myocardial ischemia-reperfusion damage in nondiabetic rats, whether vasonatrin peptide (VNP), the artificial synthetic chimera of atrial natriuretic peptide and C-type natriuretic peptide, confers cardioprotective effects against ischemia-reperfusion injury, especially in diabetic patients, is still unclear. This study was designed to investigate the effects of VNP on ischemia-reperfusion injury in diabetic rats and to further elucidate its mechanisms. The high-fat diet-fed streptozotocin-induced diabetic Sprague-Dawley rats were subjected to ischemia-reperfusion operation. VNP treatment (100 μg/kg iv, 10 min before reperfusion) significantly improved the instantaneous first derivation of left ventricle pressure (±LV dP/dtmax) and LV systolic pressure and reduced LV end-diastolic pressure, apoptosis index, caspase-3 activity, plasma creatine kinase (CK), and lactate dehydrogenase (LDH) activities. Moreover, VNP inhibited endoplasmic reticulum (ER) stress by suppressing glucose-regulated protein 78 (GRP78) and C/EBP homologous protein (CHOP). These effects were mimicked by 8-bromine-cyclic guanosinemonophosphate (8-Br-cGMP), a cGMP analog, whereas they were inhibited by KT-5823, the selective inhibitor of PKG. In addition, pretreatment with tauroursodeoxycholic acid (TUDCA), a specific inhibitor of ER stress, could not further promote the VNP's cardioprotective effect in diabetic rats. In vitro H9c2 cardiomyocytes were subjected to hypoxia/reoxygenation and incubated with or without VNP (10(-8) mol/l). Gene knockdown of PKG1α with siRNA blunted VNP inhibition of ER stress and apoptosis, while overexpression of PKG1α resulted in significant decreased ER stress and apoptosis. VNP protects the diabetic heart against ischemia-reperfusion injury by inhibiting ER stress via the cGMP-PKG signaling pathway. These results suggest that VNP may have potential therapeutic value for the diabetic patients with ischemic heart disease.

The Effect of Xenon on Isoflurane Protection Against Experimental Myocardial Infarction

To investigate if the protective effects of xenon and isoflurane against myocardial ischemia-reperfusion damage would be additive. A prospective, randomized laboratory investigation. An animal laboratory of a university hospital. Thirty-six pigs (female German landrace). In an open-chest preparation with thiopental anesthesia, the left anterior descending artery was occluded to produce ischemia for 60 minutes. One hour previously, ischemic preconditioning, isoflurane (0.55 minimum alveolar concentration [MAC]) alone, or isoflurane together with xenon (0.55 MAC each) were started in the respective groups. A fourth (control) group received no protective intervention. Myocardial ischemia was followed by 2 hours of reperfusion. Hearts were excised and stained (Evans Blue/TTC) to measure infarct size as related to the area at risk. Myocardial infarct size was reduced (means +/- standard deviation) from 64% +/- 9% of the area at risk in the control group to 19% +/- 12% with ischemic preconditioning to 46% +/- 12% with isoflurane and to 39% +/- 13% with isoflurane and xenon. All intervention groups were significantly different from the control (p < 0.05), and both anesthetic groups were significantly different from ischemic preconditioning (p < 0.05). Combined isoflurane/xenon anesthesia reduced infarct size but not more than isoflurane alone. Ischemic preconditioning was more effective than the anesthetics.

In vivo protein transduction: delivery of PEP-1-SOD1 fusion protein into myocardium efficiently protects against ischemic insult.

Myocardial ischemia-reperfusion injury is a medical problem occurring as damage to the myocardium following blood flow restoration after a critical period of coronary occlusion. Oxygen free radicals (OFR) are implicated in reperfusion injury after myocardial ischemia. The antioxidant enzyme, Cu, Zn-superoxide dismutase (Cu, Zn-SOD, also called SOD1) is one of the major means by which cells counteract the deleterious effects of OFR after ischemia. Recently, we reported that a PEP-1-SOD1 fusion protein was efficiently delivered into cultured cells and isolated rat hearts with ischemia-reperfusion injury. In the present study, we investigated the protective effects of the PEP-1-SOD1 fusion protein after ischemic insult. Immunofluorescecnce analysis revealed that the expressed and purified PEP-1-SOD1 fusion protein injected into rat tail veins was efficiently transduced into the myocardium with its native protein structure intact. When injected into Sprague-Dawley rat tail veins, the PEP-1- SOD1 fusion protein significantly attenuated myocardial ischemia-reperfusion damage; characterized by improving cardiac function of the left ventricle, decreasing infarct size, reducing the level of malondialdehyde (MDA), decreasing the release of creatine kinase (CK) and lactate dehydrogenase (LDH), and relieving cardiomyocyte apoptosis. These results suggest that the biologically active intact forms of PEP-1-SOD1 fusion protein will provide an efficient strategy for therapeutic delivery in various diseases related to SOD1 or to OFR.

Reduction of nitrite to nitric oxide during ischemia protects against myocardial ischemia-reperfusion damage.

Nitric oxide (NO.) is thought to protect against the damaging effects of myocardial ischemia-reperfusion injury, whereas xanthine oxidoreductase (XOR) normally causes damage through the generation of reactive oxygen species. In the heart, inorganic nitrite (NO(2)(-)) has the potential to act as an endogenous store of NO., liberated specifically during ischemia. Using a detection method that we developed, we report that under ischemic conditions both rat and human homogenized myocardium and the isolated perfused rat heart (Langendorff preparation) generate NO. from NO(2)(-) in a reaction that depends on XOR activity. Functional studies of rat hearts in the Langendorff apparatus showed that nitrite (10 and 100 microM) reduced infarct size from 47.3 +/- 2.8% (mean percent of control +/- SEM) to 17.9 +/- 4.2% and 17.4 +/- 1.0%, respectively (P < 0.001), and was associated with comparable improvements in recovery of left ventricular function. This protective effect was completely blocked by the NO. scavenger 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazole-1-oxyl 3-oxide (carboxy-PTIO). In summary, the generation of NO. from NO(2)(-), by XOR, protects the myocardium from ischemia-reperfusion injury. Hence, if XOR is presented with NO(2)(-) as an alternative substrate, the resultant effects of its activity may be protective, by means of its production of NO. , rather than damaging.

Myocardial ischemia-reperfusion damage after pacing-induced tachycardia in patients with cardiac syndrome X

The presence of myocardial ischemia in syndrome X (chest pain, "ischemia-like" electrocardiogram changes, and normal coronary angiograms) is uncertain possibly because, when focally distributed, it may not cause contractile dysfunction or lactate production. We measured lipid hydroperoxides (ROOHs) and conjugated dienes (CDs), two sensitive, independent markers of ischemia-reperfusion oxidative stress, in paired aortic and great cardiac vein blood samples before and after pacing-induced tachycardia in nine patients with syndrome X. Diagnostic ischemic S-T segment changes during pacing were followed by a consistent increase in ROOH and CD levels in the great cardiac vein (from 4.83 +/- 1.18 micromol/l at baseline to 7.88 +/- 1.12 micromol/l and from 0.038 +/- 0.002 to 0.051 +/- 0.003 arbitrary units, respectively, P < 0.01). In controls, ROOH and CD levels did not change after pacing. The large postpacing cardiac release of lipid peroxidation products, consistently observed in all patients and similar to that previously observed after ischemia caused by percutaneous transluminal coronary angioplasty, is consistent with an ischemic origin of syndrome X.

Ischemic heart disease and antioxidants: mechanistic aspects of oxidative injury and its prevention.

The disease state of myocardial ischemia results from a hypoperfusion-induced insufficiency of heart-muscle oxidative metabolism due to inadequate coronary circulation. Myocardial ischemia is an important, lifespan-limiting medical problem and a major economic health-care concern. Reperfusion, although avidly pursued in the clinic as essential to the ultimate survival of acutely ischemic heart muscle, may itself carry an injury component. Cardiac reperfusion injury appears to reflect, at least in part, an oxidant burden established upon reoxygenation of ischemic myocardium. Laboratory evidence demonstrates that oxidative stress to the heart-muscle cell (cardiomyocyte) can elicit the three known types of ischemia-reperfusion injury that directly affect the myocardium: arrhythmia, stunning, and infarction. The limited clinical occurrence of serious reperfusion arrhythmias has restricted the importance of antioxidants as antiarrhythmic agents against this form of myocardial ischemia-reperfusion damage. Despite the utmost clinical significance of lethal cardiomyocyte injury as a negative prognostic indicator for the ischemic heart-disease patient, inconsistent results of antioxidant interventions in reducing infarct size have somewhat tempered interest in antioxidant infarct trials. By contrast, the negative clinical consequences of stunning may indeed be preventable by utilizing antioxidants to help restore postischemic cardiac pump function. Several as yet unanswered questions remain regarding oxidative stress in the reperfused heart, its significance to cardiomyocyte damage, and its ability to elicit specific postischemic myocardial derangements. Targeted mechanistic studies are required to address these questions and to define the pathogenic role of oxidative stress (and, hence, the therapeutic potential of antioxidant intervention) in myocardial ischemia-reperfusion injury. The overall aim of current research in this area is to enable the cardiac surgeon/cardiologist to advance beyond the largely palliative drugs now available for management of the coronary heart-disease patient and attack directly the pathogenic determinants of heart-muscle ischemia-reperfusion injury. Optimal use of antioxidants may help address this important medical need.