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Poster De Conférence Année : 2022

Hypoxia-reoxygenation induces neutral lipid accumulation in cardiomyocytes printemp

Résumé

Introduction Circulating lipids are well established cardiovascular risk factors but, independently from dyslipidemia, their direct physiopathological role on cardiomyocytes is poorly known. We previously demonstrated that reperfusion of an ischemic myocardium induced accumulation of both cholesterol and oxysterols in mitochondria. Pharmacological inhibition of this phenomenon with the use of 4'chlorodiazepam protected mitochondrial function and reduced infarct size. Nevertheless, the distribution and the trafficking of lipids in cardiomyocytes, especially cholesterol, during cardiac ischemia-reperfusion is still ill known. Objective The goal of this study is to investigate the dynamic of lipids during hypoxia-reoxygenation in a cellular model. Method AC16 human cardiomyocytes were submitted to 14h hypoxia (1% O2) followed by 2h reoxygenation (21% O2). Mortality was assessed with crystal violet assay. Cytosolic lipid droplet formation and plasma membrane cholesterol were monitored by fluorescent microscopy using Bodipy and D4-mCherry probes, respectively. Whole cell neutral lipids were quantified by thin layer chromatography. Results 14h hypoxia followed by 2h reoxygenation induced 41±3% of cell mortality compared to normoxia, (n=5, p<0.05). Hypoxiareoxygenation also induced lipid droplet formation (+2368±878% vs. normoxia, n=4, p<0.05) and increased cholesterol-rich domains in plasma membrane (258±29% vs. normoxia, n=5, p<0.05). These results were confirmed by thin layer chromatography, showing an increase in triglycerides, cholesterol esters and free fatty acid in hypoxia-reoxygenation compared to normoxia (+291±30%, +657±306% and +315±81%, respectively; n=5, p<0.05). Conclusion Hypoxia-reoxygenation induces cytosolic neutral lipid and plasma membrane cholesterol accumulation in cardiomyocytes. Whether this change of lipid distribution is responsible for mitochondrial cholesterol accumulation previously described at reperfusion needs to be determined.
2022 GRRC support final Juliette Bréhat.pdf (5.58 Mo) Télécharger le fichier
2022 GRRC Abstract final Juliette Bréhat.pdf (43.91 Ko) Télécharger le fichier
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Dates et versions

hal-03828113 , version 1 (13-04-2023)

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  • HAL Id : hal-03828113 , version 1

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Juliette Bréhat, Alexis Nguyen, Leeyah Issop, Nario Tomishige, Toshihide Kobayashi, et al.. Hypoxia-reoxygenation induces neutral lipid accumulation in cardiomyocytes printemp. Printemps de la cardiologie, Jun 2022, Tours, France. ⟨hal-03828113⟩
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