Effect of RONS-Induced Intracellular Redox Homeostasis in 6-NBDG/Glucose Uptake in C2C12 Myotubes and Single Isolated Skeletal Muscle Fibres
The glucose uptake in skeletal muscle is essential to produce energy through ATP, which is needed by this organ to maintain vital functions. The impairment of glucose uptake compromises the metabolism and function of skeletal muscle and other organs and is a feature of diabetes, obesity, and ageing....
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-04-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/24/9/8082 |
| _version_ | 1797602482607620096 |
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| author | Escarlata Fernández-Puente Eva Martín-Prieto Carlos Manuel Márquez Jesús Palomero |
| author_facet | Escarlata Fernández-Puente Eva Martín-Prieto Carlos Manuel Márquez Jesús Palomero |
| author_sort | Escarlata Fernández-Puente |
| collection | DOAJ |
| description | The glucose uptake in skeletal muscle is essential to produce energy through ATP, which is needed by this organ to maintain vital functions. The impairment of glucose uptake compromises the metabolism and function of skeletal muscle and other organs and is a feature of diabetes, obesity, and ageing. There is a need for research to uncover the mechanisms involved in the impairment of glucose uptake in skeletal muscle. In this study, we adapted, developed, optimised, and validated a methodology based on the fluorescence glucose analogue 6-NBDG, combined with a quantitative fluorescence microscopy image analysis, to determine the glucose uptake in two models of skeletal muscle cells: C2C12 myotubes and single fibres isolated from muscle. It was proposed that reactive oxygen and nitrogen species (RONS) and redox homeostasis play an important role in the modulation of intracellular redox signalling pathways associated with glucose uptake. In this study, we prove that the prooxidative intracellular redox environment under oxidative eustress produced by RONS such as hydrogen peroxide and nitric oxide improves glucose uptake in skeletal muscle cells. However, when oxidation is excessive, oxidative distress occurs, and cellular viability is compromised, although there might be an increase in the glucose uptake. Based on the results of this study, the determination of 6-NBDG/glucose uptake in myotubes and skeletal muscle cells is feasible, validated, and will contribute to improve future research. |
| first_indexed | 2024-03-11T04:17:38Z |
| format | Article |
| id | doaj.art-ec7074bb5d854c99925d3cba5f698a68 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-11T04:17:38Z |
| publishDate | 2023-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-ec7074bb5d854c99925d3cba5f698a682023-11-17T23:04:40ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-04-01249808210.3390/ijms24098082Effect of RONS-Induced Intracellular Redox Homeostasis in 6-NBDG/Glucose Uptake in C2C12 Myotubes and Single Isolated Skeletal Muscle FibresEscarlata Fernández-Puente0Eva Martín-Prieto1Carlos Manuel Márquez2Jesús Palomero3Department of Physiology and Pharmacology, University of Salamanca, 37007 Salamanca, SpainDepartment of Physiology and Pharmacology, University of Salamanca, 37007 Salamanca, SpainDepartment of Physiology and Pharmacology, University of Salamanca, 37007 Salamanca, SpainDepartment of Physiology and Pharmacology, University of Salamanca, 37007 Salamanca, SpainThe glucose uptake in skeletal muscle is essential to produce energy through ATP, which is needed by this organ to maintain vital functions. The impairment of glucose uptake compromises the metabolism and function of skeletal muscle and other organs and is a feature of diabetes, obesity, and ageing. There is a need for research to uncover the mechanisms involved in the impairment of glucose uptake in skeletal muscle. In this study, we adapted, developed, optimised, and validated a methodology based on the fluorescence glucose analogue 6-NBDG, combined with a quantitative fluorescence microscopy image analysis, to determine the glucose uptake in two models of skeletal muscle cells: C2C12 myotubes and single fibres isolated from muscle. It was proposed that reactive oxygen and nitrogen species (RONS) and redox homeostasis play an important role in the modulation of intracellular redox signalling pathways associated with glucose uptake. In this study, we prove that the prooxidative intracellular redox environment under oxidative eustress produced by RONS such as hydrogen peroxide and nitric oxide improves glucose uptake in skeletal muscle cells. However, when oxidation is excessive, oxidative distress occurs, and cellular viability is compromised, although there might be an increase in the glucose uptake. Based on the results of this study, the determination of 6-NBDG/glucose uptake in myotubes and skeletal muscle cells is feasible, validated, and will contribute to improve future research.https://www.mdpi.com/1422-0067/24/9/8082glucose uptake6-NBDGinsulin resistanceC2C12 myotubesskeletal muscle fibresROS |
| spellingShingle | Escarlata Fernández-Puente Eva Martín-Prieto Carlos Manuel Márquez Jesús Palomero Effect of RONS-Induced Intracellular Redox Homeostasis in 6-NBDG/Glucose Uptake in C2C12 Myotubes and Single Isolated Skeletal Muscle Fibres International Journal of Molecular Sciences glucose uptake 6-NBDG insulin resistance C2C12 myotubes skeletal muscle fibres ROS |
| title | Effect of RONS-Induced Intracellular Redox Homeostasis in 6-NBDG/Glucose Uptake in C2C12 Myotubes and Single Isolated Skeletal Muscle Fibres |
| title_full | Effect of RONS-Induced Intracellular Redox Homeostasis in 6-NBDG/Glucose Uptake in C2C12 Myotubes and Single Isolated Skeletal Muscle Fibres |
| title_fullStr | Effect of RONS-Induced Intracellular Redox Homeostasis in 6-NBDG/Glucose Uptake in C2C12 Myotubes and Single Isolated Skeletal Muscle Fibres |
| title_full_unstemmed | Effect of RONS-Induced Intracellular Redox Homeostasis in 6-NBDG/Glucose Uptake in C2C12 Myotubes and Single Isolated Skeletal Muscle Fibres |
| title_short | Effect of RONS-Induced Intracellular Redox Homeostasis in 6-NBDG/Glucose Uptake in C2C12 Myotubes and Single Isolated Skeletal Muscle Fibres |
| title_sort | effect of rons induced intracellular redox homeostasis in 6 nbdg glucose uptake in c2c12 myotubes and single isolated skeletal muscle fibres |
| topic | glucose uptake 6-NBDG insulin resistance C2C12 myotubes skeletal muscle fibres ROS |
| url | https://www.mdpi.com/1422-0067/24/9/8082 |
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