The Effects of Different Fluorescent Indicators in Observing the Changes of the Mitochondrial Membrane Potential during Oxidative Stress-Induced Mitochondrial Injury of Cardiac H9c2 Cells

Sun, Yahan; Zhou, Kang; He, Mali; Gao, Ying; Zhang, Danjie; Bai, Yanwen; Lai, Yuezhao; Liu, Mengying; Han, Xuechao; Xu, Sen; Tian, Wei; Xu, Jingman

doi:10.1007/s10895-020-02623-x

Cited by 11 publications

(4 citation statements)

References 19 publications

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“…Rhodamine 123, a lipophilic cationic fluorescent substance, can enter the cell and reach the cell matrix depending on the transmembrane potential (Sun et al, 2020). Depolarization of cell membrane will result in the leakage of Rhodamine 123 and the rapid decrease of mean fluorescence intensity.…”

Section: T-atpase Activitiesmentioning

confidence: 99%

Antimicrobial activity and mechanism of limonene against Staphylococcus aureus

Han

Chen

Sun

2021

Journal of Food Safety

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In this study, inhibitory effect of limonene on Staphylococcus aureus (S. aureus) and its antibacterial mechanism was explored, respectively. The results showed that limonene effectively inhibited bacterial growth at a minimum inhibitory concentration (MIC) of 20 ml/L. Scanning electron microscopy (SEM), the reduction of AKP activity and fluorescence microscope observation confirmed that limonene caused the destruction of the cell morphology and cell wall integrity of S. aureus. The reduction of MFI in fluorescein diacetate staining experiment and leakage of biological macromolecules (nucleic acids and proteins) indicated that limonene damaged the cell membrane and increased membrane permeability. Besides, the reduction of membrane potential (MP) further confirmed that the damage to membrane and the reduction of respiratory metabolic activity. Furthermore, the respiratory depression test confirmed that the pathway of limonene affecting the respiratory metabolism of S. aureus is TCA pathway, followed by EMP. In addition, the disorder of succinate dehydrogenase (SDH), malate dehydrogenase (MDH), pyruvate kinase (PK), ATPase (T-ATPase, Na + -K + -ATPase, Ca 2+ -Mg 2+ -ATPase), and ATP concentration demonstrated that limonene could result in a slowdown of metabolism and metabolic dysfunction, thus inhibiting ATP synthesis. | INTRODUCTIONFood-borne pathogens are the leading cause of food-borne illness and one of the common pathogens causing food safety risks, causing great harm to human health (Qi et al., 2020;Tomao, Pirolo, Agnoletti, Pantosti, & Visca, 2020). Staphylococcus aureus (S. aureus), a Gram-positive bacterium with a arranged in spherical clusters shape under the microscope (Muniz et al., ), widely distributed in meat products (Rb et al., 2020), frozen food, and seafood, (Saklani, Lekshmi, Nayak, & Kumar, 2020). Staphylococcal enterotoxins are the causative toxins produced by S. aureus that can cause cutaneous lesions, pneumonia,

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Section: T-atpase Activitiesmentioning

confidence: 99%

Antimicrobial activity and mechanism of limonene against Staphylococcus aureus

Han

Chen

Sun

2021

Journal of Food Safety

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“…Here, TUNEL and JC-1 staining revealed that the protective effect of HSYA against MI/RI is related to the inhibition of apoptosis. JC-1 staining reflects the mitochondrial membrane potential, which also suggests that HSYA has a regulatory effect on mitochondrial function [37]. Cardiomyocytes are responsible for the main cardiac task of excitationcontraction coupling [38].…”

Section: Discussionmentioning

confidence: 99%

Hydroxysafflor Yellow A Ameliorates Myocardial Ischemia/Reperfusion Injury by Suppressing Calcium Overload and Apoptosis

Wang

et al. 2021

Oxidative Medicine and Cellular Longevity

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Myocardial ischemia/reperfusion injury (MI/RI) is an urgent problem with a great impact on health globally. However, its pathological mechanisms have not been fully elucidated. Hydroxysafflor yellow A (HSYA) has a protective effect against MI/RI. This study is aimed at further clarifying the relationship between HSYA cardioprotection and calcium overload as well as the underlying mechanisms. We verified the protective effect of HSYA on neonatal rat primary cardiomyocytes (NPCMs) and human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) from hypoxia-reoxygenation (HR) injury. To explore the cardioprotective mechanism of HSYA, we employed calcium fluorescence, TUNEL assay, JC-1 staining, and western blotting. Finally, cardio-ECR and patch-clamp experiments were used to explain the regulation of L-type calcium channels (LTCC) in cardioprotection mediated by HSYA. The results showed that HSYA reduced the levels of myocardial enzymes and protected NPCMs from HR injury. HSYA also restored the contractile function of hiPSC-CMs and field potential signal abnormalities caused by HR and exerted a protective effect on cardiac function. Further, we demonstrated that HSYA protects cardiomyocytes from HR injury by decreasing mitochondrial membrane potential and inhibiting apoptosis and calcium overload. Patch-clamp results revealed that MI/RI caused a sharp increase in calcium currents, which was inhibited by pretreatment with HSYA. Furthermore, we found that HSYA restored contraction amplitude, beat rate, and field potential duration of hiPSC-CMs, which were disrupted by the LTCC agonist Bay-K8644. Patch-clamp experiments also showed that HSYA inhibits Bay-K8644-induced calcium current, with an effect similar to that of the LTCC inhibitor nisoldipine. Therefore, our data suggest that HSYA targets LTCC to inhibit calcium overload and apoptosis of cardiomyocytes, thereby exerting a cardioprotective effect and reducing MI/RI injury.

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“…Cells were cultured and treated for 4 days according to the protocol for cell growth measurements. On day 4, mitochondrial membrane potential was analyzed using the cationic dye tetramethylrhodamine ethyl ester perchlorate (TMRE, Cayman Chemical, Ann Arbour, Michigan, USA) as previously described 63 . Briefly, a TMRE stock solution was prepared at a concentration of 20 µM in sterile dimethyl sulfoxide and stored at − 20 °C.…”

Section: Methodsmentioning

confidence: 99%

N-oleoylethanolamide treatment of lymphoblasts deficient in Tafazzin improves cell growth and mitochondrial morphology and dynamics

Chan

Fernandes

Steckel

et al. 2022

Sci Rep

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Barth syndrome (BTHS) is caused by mutations in the TAZ gene encoding the cardiolipin remodeling enzyme, Tafazzin. The study objective was to quantitatively examine growth characteristics and mitochondrial morphology of transformed lymphoblast cell lines derived from five patients with BTHS relative to five healthy controls, as well as the therapeutic potential of oleoylethanolamide (OEA) and linoleoylethanolamide (LEA). These bioactive lipids both activate PPARα, which may be therapeutic. BTHS lymphoblasts grew more slowly than controls, suggesting lymphopenia merits clinical investigation. Treatment of BTHS lymphoblasts with OEA, but not LEA, significantly restored mitochondrial membrane potential, as well as colony growth in all BTHS lymphoblast lines, although a full growth rescue was not achieved. Quantification analysis of electron micrographs from three BTHS and healthy lymphoblast donors indicated similar numbers of mitochondria per cell, but lower average cristae length per mitochondrion, and higher mitochondrial density. Additionally, BTHS lymphoblasts had larger mitochondria, and a higher percentage of abnormally large mitochondria (> 1 μm2) than healthy controls. Notably, OEA treatment significantly restored mitochondrial size, without affecting density or cristae lengths. Cardiolipin total content, relative linoleic acid content and monolysocardiolipin:cardiolipin ratios were not improved by OEA, indicating that effects on growth, and mitochondrial morphology and function, occurred without resolving this deficit. However, immunoblotting showed higher levels of OPA1, a biomarker for mitochondrial fusion, in BTHS lymphoblasts, which was attenuated by OEA treatment, implicating altered mitochondrial dynamics in the pathology and treatment of BTHS.

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The Effects of Different Fluorescent Indicators in Observing the Changes of the Mitochondrial Membrane Potential during Oxidative Stress-Induced Mitochondrial Injury of Cardiac H9c2 Cells

Cited by 11 publications

References 19 publications

Antimicrobial activity and mechanism of limonene against Staphylococcus aureus

Antimicrobial activity and mechanism of limonene against Staphylococcus aureus

Hydroxysafflor Yellow A Ameliorates Myocardial Ischemia/Reperfusion Injury by Suppressing Calcium Overload and Apoptosis

N-oleoylethanolamide treatment of lymphoblasts deficient in Tafazzin improves cell growth and mitochondrial morphology and dynamics

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