Bioreactor-based 3D human myocardial ischemia/reperfusion in vitro model: a novel tool to unveil key paracrine factors upon acute myocardial infarction

Cardiovascular disease is the leading cause of death all over the world: for no other reason as many people die every year as from the cardiovascular disease. Taking into account the high medical and social significance of the problem of treating patients with coronary heart disease and acute myocardial infarction, the search for new effective methods of preventing or weakening ischemic myocardial damage and revealing the mechanisms of their implementation is an urgent task of modern experimental and clinical medicine. The aim of the study was to experimentally test the cardioprotective efficiency of pharmacological postconditioning with lactic acid in myocardial ischemia-reperfusion in rats with transient hypercholesterolemia.In the course of the study, it was found that remote ischemic postconditioning (RIPostC) during myocardial ischemia-reperfusion leads to an increase in the level of lactate in the blood and has an infarction-limiting effect in rats. It was revealed that lactate, after injection into the bloodstream of animals at a dose of 10 mg/kg 25 minutes after the start of reperfusion, like RIPostC, leads to a decrease in the necrosis area in the left ventricular of the myocardium. Pharmacological postconditioning with lactate is ineffective in the limiting necrosis area in the myocardium of the left ventricle of rats with transient hypercholesterolemia. The presence of such a risk factor for cardiovascular diseases as hypercholesterolemia can serve as a criterion for excluding the use of pharmacological postconditioning with lactate as a way to reduce ischemic and reperfusion damage to the myocardium in patients with acute myocardial infarction.

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iPLA2β Contributes to ER Stress-Induced Apoptosis during Myocardial Ischemia/Reperfusion Injury

Cells ◽

10.3390/cells10061446 ◽

2021 ◽

Vol 10 (6) ◽

pp. 1446

Author(s):

Tingting Jin ◽

Jun Lin ◽

Yingchao Gong ◽

Xukun Bi ◽

Shasha Hu ◽

...

Keyword(s):

Cell Death ◽

Heart Disease ◽

Myocardial Ischemia ◽

Ischemic Heart Disease ◽

Er Stress ◽

Ischemia Reperfusion ◽

Myocardial Ischemia Reperfusion ◽

Induced Apoptosis

Both calcium-independent phospholipase A2 beta (iPLA2β) and endoplasmic reticulum (ER) stress regulate important pathophysiological processes including inflammation, calcium homeostasis and apoptosis. However, their roles in ischemic heart disease are poorly understood. Here, we show that the expression of iPLA2β is increased during myocardial ischemia/reperfusion (I/R) injury, concomitant with the induction of ER stress and the upregulation of cell death. We further show that the levels of iPLA2β in serum collected from acute myocardial infarction (AMI) patients and in samples collected from both in vivo and in vitro I/R injury models are significantly elevated. Further, iPLA2β knockout mice and siRNA mediated iPLA2β knockdown are employed to evaluate the ER stress and cell apoptosis during I/R injury. Additionally, cell surface protein biotinylation and immunofluorescence assays are used to trace and locate iPLA2β. Our data demonstrate the increase of iPLA2β augments ER stress and enhances cardiomyocyte apoptosis during I/R injury in vitro and in vivo. Inhibition of iPLA2β ameliorates ER stress and decreases cell death. Mechanistically, iPLA2β promotes ER stress and apoptosis by translocating to ER upon myocardial I/R injury. Together, our study suggests iPLA2β contributes to ER stress-induced apoptosis during myocardial I/R injury, which may serve as a potential therapeutic target against ischemic heart disease.

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Minocycline induces protective autophagy in vascular endothelial cells exposed to an in vitro model of ischemia/reperfusion-induced injury

Biomedical Reports ◽

10.3892/br.2015.554 ◽

2015 ◽

Vol 4 (2) ◽

pp. 173-177 ◽

Cited By ~ 13

Author(s):

WENBIN DONG ◽

SHIGENG XIAO ◽

MIN CHENG ◽

XIAODI YE ◽

GAOLI ZHENG

Keyword(s):

Endothelial Cells ◽

In Vitro Model ◽

Vascular Endothelial Cells ◽

Ischemia Reperfusion ◽

Vascular Endothelial ◽

Vitro Model

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Abstract 423: Proteasome Priming by Protein Kinase G Protects Against Myocardial Ischemia-reperfusion Injury

Circulation Research ◽

10.1161/res.117.suppl_1.423 ◽

2015 ◽

Vol 117 (suppl_1) ◽

Author(s):

Xuejun Wang ◽

Erin J Terpstra ◽

Eduardo Callegari ◽

Chengjun Hu ◽

Hanming Zhang ◽

...

Keyword(s):

Protein Kinase ◽

Myocardial Ischemia ◽

Ischemia Reperfusion Injury ◽

Ischemia Reperfusion ◽

Underlying Mechanism ◽

26S Proteasome ◽

Transgenic Mouse Line ◽

Pde5 Inhibition ◽

Myocardial Ischemia Reperfusion

Cardiac proteasome functional insufficiency is implicated in a large subset of heart disease and has been experimentally demonstrated to play an essential role in cardiac proteotoxicity, including desmin-related cardiomyopathy and myocardial ischemia-reperfusion (I-R) injury. Pharmacological inhibition of phosphodiesterase 5 (PDE5) via sildenafil for example, which can stabilize cGMP and thereby increase cGMP-dependent protein kinase (PKG) activity, is consistently reported to protect against I-R injury; however, the underlying mechanism is not fully understood. We have recently discovered that PKG activation enhances proteasomal degradation of misfolded proteins (Ranek, et al. Circulation 2013), prompting us to hypothesize that proteasome-priming may contribute to cardioprotection-induced by PDE5 inhibition. Here we used a cardiomyocyte-restricted proteasome inhibition transgenic mouse line (Tg) and non-Tg (Ntg) littermates to interrogate the action of sildenafil on I-R injury created by left anterior descending artery (LAD) ligation (30 min) and release (24 hr). Sildenafil was administered 30 min before LAD ligation. Results showed that (1) the 26S proteasome activity of the Ntg I-R hearts was significantly elevated by sildenafil but this elevation was blocked in the Tg line; (2) the infarct size reduction by sildenafil treatment in Ntg mice was completely abolished in the Tg mice with the same treatment; and (3) systolic and diastolic function impairment after I/R was markedly attenuated in sildenafil-treated Ntg mice, but not in the sildenafil-treated Tg mice. Additionally, immunoprecipitation assays show that PKG interacted with the proteasome in cultured cardiomyocytes, and this interaction appeared to be augmented by sildenafil treatment. Moreover, in vitro incubation of active PKG with purified human 26S proteasomes increased proteasome peptidase activities and the phosphorylation at specific serine residues of a 19S proteasome subunit as revealed by “gel-free” nano-LC-MS/MS. We conclude that active PKG directly interacts with, phosphorylates, and increases the activities of, the proteasome and that proteasome priming mediates to cardioprotection of PDE5 inhibition against I-R injury.

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Trop2 Guarantees Cardioprotective Effects of Cortical Bone-Derived Stem Cells on Myocardial Ischemia/Reperfusion Injury

Cell Transplantation ◽

10.1177/0963689718786882 ◽

2018 ◽

Vol 27 (8) ◽

pp. 1256-1268 ◽

Cited By ~ 1

Author(s):

Tianyu Li ◽

Yunshu Su ◽

Xiongli Yu ◽

Durgahee S.A. Mouniir ◽

Jackson Ferdinand Masau ◽

...

Keyword(s):

Myocardial Infarction ◽

Stem Cells ◽

Myocardial Ischemia ◽

Cortical Bone ◽

Heart Function ◽

Heart Diseases ◽

Ischemia Reperfusion ◽

Promising Therapeutic Approach ◽

Cardioprotective Effects ◽

Myocardial Ischemia Reperfusion

Stem cell transplantation represents a promising therapeutic approach for myocardial ischemia/reperfusion (I/R) injury, where cortical bone-derived stem cells (CBSCs) stand out and hold superior cardioprotective effects on myocardial infarction than other types of stem cells. However, the molecular mechanism underlying CBSCs function on myocardial I/R injury is poorly understood. In a previous study, we reported that Trop2 (trophoblast cell-surface antigen 2) is expressed exclusively on the CBSCs membrane, and is involved in regulation of proliferation and differentiation of CBSCs. In this study, we found that the Trop2 is essential for the ameliorative effects of CBSCs on myocardial I/R-induced heart damage via promoting angiogenesis and inhibiting cardiomyocytes apoptosis in a paracrine manner. Trop2 is required for the colonization of CBSCs in recipient hearts. When Trop2 was knocked out, CBSCs largely lost their functions in lowering myocardial infarction size, improving heart function, enhancing capillary density, and suppressing myocardial cell death. Mechanistically, activating the AKT/GSK3β/β-Catenin signaling axis contributes to the essential role of Trop2 in CBSCs-rendered cardioprotective effects on myocardial I/R injury. In conclusion, maintaining the expression and/or activation of Trop2 in CBSCs might be a promising strategy for treating myocardial infarction, I/R injury, and other related heart diseases.

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Effects of Ganoderma Lucidum on Contractile Dysfunction of the Rabbit Urinary Bladder using an In-Vitro Model of Ischemia/Reperfusion

The Natural Products Journal ◽

10.2174/2210315507666171004160713 ◽

2018 ◽

Vol 8 (1) ◽

pp. 77-83

Author(s):

Robert M. Levin ◽

Li-Xia ◽

Wu Wei ◽

Catherine Schuler ◽

Robert E. Leggett ◽

...

Keyword(s):

Urinary Bladder ◽

Ganoderma Lucidum ◽

In Vitro Model ◽

Ischemia Reperfusion ◽

Contractile Dysfunction ◽

Vitro Model ◽

Rabbit Urinary Bladder

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

Frontiers in Cardiovascular Medicine ◽

10.3389/fcvm.2021.747802 ◽

2021 ◽

Vol 8 ◽

Author(s):

Cuizhi Li ◽

Huafeng Song ◽

Chunlin Chen ◽

Shaoxian Chen ◽

Qiyu Zhang ◽

...

Keyword(s):

Myocardial Ischemia ◽

Cell Viability ◽

Ischemia Reperfusion ◽

Inflammatory Factors ◽

H9c2 Cells ◽

Myocardial Ischemia Reperfusion ◽

Tissue Specimens ◽

Masson Staining

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.

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