Mitochondrial proteomics alterations in rat hearts following ischemia/reperfusion and diazoxide post‑conditioning

Pan, Yunchao; Wang, Yuan; Shi, Wen‐Yan; Liu, Yun; Cao, Song; Yu, Tian

doi:10.3892/mmr.2020.11800

Cited by 8 publications

(7 citation statements)

References 31 publications

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“…The results showed that diazoxide post-conditioning significantly elevates the protein expression levels of HIF-1α, VEGF, and bcl-2, leading to improvements in the myocardial ultrastructure, mitochondrial Flameng score, and membrane potential. In addition, our previous research has shown that inhibition of the HIF-1/HRE pathway by the HIF-1α subunit blocker, 2ME2, can eliminate the myocardial protective effects of diazoxide post-conditioning in isolated rat hearts ( 25 , 26 ). On this basis, we have confirmed that the myocardial protective effects of diazoxide post-conditioning can be attributed to upregulation of the HIF-1/HRE pathway.…”

Section: Discussionmentioning

confidence: 99%

Diazoxide Post-conditioning Activates the HIF-1/HRE Pathway to Induce Myocardial Protection in Hypoxic/Reoxygenated Cardiomyocytes

Chen

Wang

Cheng

et al. 2021

Front. Cardiovasc. Med.

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Background: Previous studies have shown that diazoxide can protect against myocardial ischemia-reperfusion injury (MIRI). The intranuclear hypoxia-inducible factor-1 (HIF-1)/hypoxia-response element (HRE) pathway has been shown to withstand cellular damage caused by MIRI. It remains unclear whether diazoxide post-conditioning is correlated with the HIF-1/HRE pathway in protective effect on cardiomyocytes.Methods: An isolated cardiomyocyte model of hypoxia-reoxygenation injury was established. Prior to reoxygenation, cardiomyocytes underwent post-conditioning treatment by diazoxide, and 5-hydroxydecanoate (5-HD), N-(2-mercaptopropionyl)-glycine (MPG), or dimethyloxallyl glycine (DMOG) followed by diazoxide. At the end of reoxygenation, ultrastructural morphology; mitochondrial membrane potential; interleukin-6 (IL-6), tumor necrosis factor alpha (TNF-α), reactive oxygen species (ROS), and HIF-1α levels; and downstream gene mRNA and protein levels were analyzed to elucidate the protective mechanism of diazoxide post-conditioning.Results: Diazoxide post-conditioning enabled activation of the HIF-1/HRE pathway to induce myocardial protection. When the mitoKATP channel was inhibited and ROS cleared, the diazoxide effect was eliminated. DMOG was able to reverse the effect of ROS absence to restore the diazoxide effect. MitoKATP and ROS in the early reoxygenation phase were key to activation of the HIF-1/HRE pathway.Conclusion: Diazoxide post-conditioning promotes opening of the mitoKATP channel to generate a moderate ROS level that activates the HIF-1/HRE pathway and subsequently induces myocardial protection.

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Section: Discussionmentioning

confidence: 99%

Diazoxide Post-conditioning Activates the HIF-1/HRE Pathway to Induce Myocardial Protection in Hypoxic/Reoxygenated Cardiomyocytes

Chen

Wang

Cheng

et al. 2021

Front. Cardiovasc. Med.

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“…To assess the biological implications of the identified substrates, we compared our data to the literature on hypoxia-reoxygenation or ischemia-reperfusion injury in the heart or cardiomyocytes. − We then compared substrates identified here to those cleaved by MMP-2 in vitro. − In addition, we took into account our accompanying paper, describing the degradome changes that occur during cardiac ischemia-reperfusion injury in rat hearts (Table S2, Supporting Information).…”

Section: Resultsmentioning

confidence: 99%

N-Terminomic Identification of Intracellular MMP-2 Substrates in Cardiac Tissue

Hartley,

Bassiouni,

Roczkowsky

et al. 2024

J. Proteome Res.

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Proteases are enzymes that induce irreversible posttranslational modifications by hydrolyzing amide bonds in proteins. One of these proteases is matrix metalloproteinase-2 (MMP-2), which has been shown to modulate extracellular matrix remodeling and intracellular proteolysis during myocardial injury. However, the substrates of MMP-2 in heart tissue are limited, and lesser known are the cleavage sites. Here, we used degradomics to investigate the substrates of intracellular MMP-2 in rat ventricular extracts. First, we designed a novel, constitutively active MMP-2 fusion protein (MMP-2-Fc) that we expressed and purified from mammalian cells. Using this protease, we proteolyzed ventricular extracts and used subtiligase-mediated N-terminomic labeling which identified 95 putative MMP-2-Fc proteolytic cleavage sites using mass spectrometry. The intracellular MMP-2 cleavage sites identified in heart tissue extracts were enriched for proteins primarily involved in metabolism, as well as the breakdown of fatty acids and amino acids. We further characterized the cleavage of three of these MMP-2-Fc substrates based on the gene ontology analysis. We first characterized the cleavage of sarco/endoplasmic reticulum calcium ATPase (SERCA2a), a known MMP-2 substrate in myocardial injury. We then characterized the cleavage of malate dehydrogenase (MDHM) and phosphoglycerate kinase 1 (PGK1), representing new cardiac tissue substrates. Our findings provide insights into the intracellular substrates of MMP-2 in cardiac cells, suggesting that MMP-2 activation plays a role in cardiac metabolism.

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“…In turn, mitochondrial dysfunctions triggered by ROS and the accumulation of calcium in the mitochondrial matrix favor cell death and inflammation [ 17 ]. The activation of mitoK-ATP under stress conditions has been proposed to cause slight mitochondrial uncoupling, which prevents ROS generation through the reverse electron transport mechanism and limits calcium overload [ 6 , 7 , 21 ]. Therefore, we propose that T3 replacement favors the antioxidant activity of mitok-ATP by upregulating the subunits of the channel under stress conditions.…”

Section: Discussionmentioning

confidence: 99%

“…The opening of the ATP-sensitive mitochondrial potassium channel (mitoK-ATP) plays a pivotal antioxidant and protective role against post-IR mitochondrial dysfunction and cell loss [ 6 , 7 ]. In line with this notion, mitoK-ATP is the final effector of several cardioprotective approaches in both acute and chronic heart disease in several animal models [ 8 , 9 , 10 ].…”

Section: Introductionmentioning

confidence: 99%

Role of miR-133/Dio3 Axis in the T3-Dependent Modulation of Cardiac mitoK-ATP Expression

Canale

Nicolini

Pitto

et al. 2022

IJMS

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The opening of the ATP-sensitive mitochondrial potassium channel (mitok-ATP) is a common goal of cardioprotective strategies in the setting of acute and chronic myocardial disease. The biologically active thyroid hormone (TH), 3-5-3-triiodothyronine (T3), has been indicated as a potential activator of mitoK-ATP but the underlying mechanisms are still elusive. Here we describe a novel role of T3 in the transcriptional regulation of mitoK and mitoSur, the recently identified molecular constituents of the channel. To mimic human ischemic heart damage, we used a rat model of a low T3 state as the outcome of a myocardial ischemia/reperfusion event, and neonatal rat cardiomyocytes (NRCM) challenged with hypoxia or H2O2. Either in the in vivo or in vitro models, T3 administration to recover the physiological concentrations was able to restore the expression level of both the channel subunits, which were found to be downregulated under the stress conditions. Furthermore, the T3-mediated transcriptional activation of mitoK-ATP in the myocardium and NRCM was associated with the repression of the TH-inactivating enzyme, deiodinase 3 (Dio3), and an up-regulation of the T3-responsive miR-133a-3p. Mechanistically, the loss and gain of function experiments and reporter gene assays performed in NRCM, have revealed a new regulatory axis whereby the silencing of Dio3 under the control of miR-133a-3p drives the T3-dependent modulation of cardiac mitoK and mitoSur transcription.

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Mitochondrial proteomics alterations in rat hearts following ischemia/reperfusion and diazoxide post‑conditioning

Cited by 8 publications

References 31 publications

Diazoxide Post-conditioning Activates the HIF-1/HRE Pathway to Induce Myocardial Protection in Hypoxic/Reoxygenated Cardiomyocytes

Diazoxide Post-conditioning Activates the HIF-1/HRE Pathway to Induce Myocardial Protection in Hypoxic/Reoxygenated Cardiomyocytes

N-Terminomic Identification of Intracellular MMP-2 Substrates in Cardiac Tissue

Role of miR-133/Dio3 Axis in the T3-Dependent Modulation of Cardiac mitoK-ATP Expression

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