Rottlerin Stimulates Exosome/Microvesicle Release Via the Increase of Ceramide Levels Mediated by Ampk in an In Vitro Model of Intracellular Lipid Accumulation
Exosomes/microvesicles originate from multivesicular bodies that allow the secretion of endolysosome components out of the cell. In the present work, we investigated the effects of rottlerin, a polyphenol, on exosome/microvesicle secretion in a model of intracellular lipid trafficking impairment, an...
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MDPI AG
2022-06-01
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| Online Access: | https://www.mdpi.com/2227-9059/10/6/1316 |
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| author | Yessenia L. Molina David García-Seisdedos Bohdan Babiy Milagros Lerma Javier Martínez-Botas María J. Casarejos María T. Vallejo Diego Gómez-Coronado Miguel A. Lasunción Óscar Pastor Rebeca Busto |
| author_facet | Yessenia L. Molina David García-Seisdedos Bohdan Babiy Milagros Lerma Javier Martínez-Botas María J. Casarejos María T. Vallejo Diego Gómez-Coronado Miguel A. Lasunción Óscar Pastor Rebeca Busto |
| author_sort | Yessenia L. Molina |
| collection | DOAJ |
| description | Exosomes/microvesicles originate from multivesicular bodies that allow the secretion of endolysosome components out of the cell. In the present work, we investigated the effects of rottlerin, a polyphenol, on exosome/microvesicle secretion in a model of intracellular lipid trafficking impairment, and elucidated the mechanism of action. In a model of lipid trafficking impairment in C6 glia cells, rottlerin increased ceramide levels, while decreasing hexosylceramide content. This was accompanied by increased exosome/microvesicle secretion, thereby reducing the concentration of lipids in the endolysosomal compartment. The reduction of hexosylceramide levels by rottlerin was attributed to the increase of β-glucosidase (glucosylceramidase) activity, and the effects of rottlerin were abrogated by β-glucosidase inhibitors such as isofagomine D-tartrate and AMP-deoxynojirimycin. Moreover, treatment with ML-266, a potent activator of the β-glucosidase enzyme, recapitulated the effects of rottlerin on the sphingolipid profile and exosome/microvesicle secretion. Finally, inhibition of AMPK (AMP-activated protein kinase) using compound C prevented both exosome/microvesicle secretion and the elimination of endolysosome lipids, which were promoted by rottlerin. The results showed that the decrease in intracellular lipid deposition induced by rottlerin was mediated by β-glucosidase activation and exosome/microvesicle release via the AMPK pathway. Rottlerin consumption could represent an additional health benefit in lysosomal deposition diseases. |
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| institution | Directory Open Access Journal |
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| language | English |
| last_indexed | 2024-03-10T00:21:11Z |
| publishDate | 2022-06-01 |
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| series | Biomedicines |
| spelling | doaj.art-c4a8ae005cca432399f9442792dac2a42023-11-23T15:42:40ZengMDPI AGBiomedicines2227-90592022-06-01106131610.3390/biomedicines10061316Rottlerin Stimulates Exosome/Microvesicle Release Via the Increase of Ceramide Levels Mediated by Ampk in an In Vitro Model of Intracellular Lipid AccumulationYessenia L. Molina0David García-Seisdedos1Bohdan Babiy2Milagros Lerma3Javier Martínez-Botas4María J. Casarejos5María T. Vallejo6Diego Gómez-Coronado7Miguel A. Lasunción8Óscar Pastor9Rebeca Busto10Servicio de Bioquímica-Investigación, Hospital Universitario Ramón y Cajal, IRYCIS, 28034 Madrid, SpainServicio de Bioquímica-Investigación, Hospital Universitario Ramón y Cajal, IRYCIS, 28034 Madrid, SpainServicio de Bioquímica-Clínica, Hospital Universitario Ramón y Cajal, IRYCIS, 28034 Madrid, SpainServicio de Bioquímica-Investigación, Hospital Universitario Ramón y Cajal, IRYCIS, 28034 Madrid, SpainServicio de Bioquímica-Investigación, Hospital Universitario Ramón y Cajal, IRYCIS, 28034 Madrid, SpainServicio de Neurobiología, Hospital Universitario Ramón y Cajal, IRYCIS, 28034 Madrid, SpainMolecular Imaging and Immunohistochemistry Unit, Hospital Universitario la Paz, IdiPAZ, 28046 Madrid, SpainServicio de Bioquímica-Investigación, Hospital Universitario Ramón y Cajal, IRYCIS, 28034 Madrid, SpainServicio de Bioquímica-Investigación, Hospital Universitario Ramón y Cajal, IRYCIS, 28034 Madrid, SpainServicio de Bioquímica-Clínica, Hospital Universitario Ramón y Cajal, IRYCIS, 28034 Madrid, SpainServicio de Bioquímica-Investigación, Hospital Universitario Ramón y Cajal, IRYCIS, 28034 Madrid, SpainExosomes/microvesicles originate from multivesicular bodies that allow the secretion of endolysosome components out of the cell. In the present work, we investigated the effects of rottlerin, a polyphenol, on exosome/microvesicle secretion in a model of intracellular lipid trafficking impairment, and elucidated the mechanism of action. In a model of lipid trafficking impairment in C6 glia cells, rottlerin increased ceramide levels, while decreasing hexosylceramide content. This was accompanied by increased exosome/microvesicle secretion, thereby reducing the concentration of lipids in the endolysosomal compartment. The reduction of hexosylceramide levels by rottlerin was attributed to the increase of β-glucosidase (glucosylceramidase) activity, and the effects of rottlerin were abrogated by β-glucosidase inhibitors such as isofagomine D-tartrate and AMP-deoxynojirimycin. Moreover, treatment with ML-266, a potent activator of the β-glucosidase enzyme, recapitulated the effects of rottlerin on the sphingolipid profile and exosome/microvesicle secretion. Finally, inhibition of AMPK (AMP-activated protein kinase) using compound C prevented both exosome/microvesicle secretion and the elimination of endolysosome lipids, which were promoted by rottlerin. The results showed that the decrease in intracellular lipid deposition induced by rottlerin was mediated by β-glucosidase activation and exosome/microvesicle release via the AMPK pathway. Rottlerin consumption could represent an additional health benefit in lysosomal deposition diseases.https://www.mdpi.com/2227-9059/10/6/1316rottlerinexosome/microvesicleintracellular lipid traffickingceramidesphingolipidlysosomal storage disease |
| spellingShingle | Yessenia L. Molina David García-Seisdedos Bohdan Babiy Milagros Lerma Javier Martínez-Botas María J. Casarejos María T. Vallejo Diego Gómez-Coronado Miguel A. Lasunción Óscar Pastor Rebeca Busto Rottlerin Stimulates Exosome/Microvesicle Release Via the Increase of Ceramide Levels Mediated by Ampk in an In Vitro Model of Intracellular Lipid Accumulation Biomedicines rottlerin exosome/microvesicle intracellular lipid trafficking ceramide sphingolipid lysosomal storage disease |
| title | Rottlerin Stimulates Exosome/Microvesicle Release Via the Increase of Ceramide Levels Mediated by Ampk in an In Vitro Model of Intracellular Lipid Accumulation |
| title_full | Rottlerin Stimulates Exosome/Microvesicle Release Via the Increase of Ceramide Levels Mediated by Ampk in an In Vitro Model of Intracellular Lipid Accumulation |
| title_fullStr | Rottlerin Stimulates Exosome/Microvesicle Release Via the Increase of Ceramide Levels Mediated by Ampk in an In Vitro Model of Intracellular Lipid Accumulation |
| title_full_unstemmed | Rottlerin Stimulates Exosome/Microvesicle Release Via the Increase of Ceramide Levels Mediated by Ampk in an In Vitro Model of Intracellular Lipid Accumulation |
| title_short | Rottlerin Stimulates Exosome/Microvesicle Release Via the Increase of Ceramide Levels Mediated by Ampk in an In Vitro Model of Intracellular Lipid Accumulation |
| title_sort | rottlerin stimulates exosome microvesicle release via the increase of ceramide levels mediated by ampk in an in vitro model of intracellular lipid accumulation |
| topic | rottlerin exosome/microvesicle intracellular lipid trafficking ceramide sphingolipid lysosomal storage disease |
| url | https://www.mdpi.com/2227-9059/10/6/1316 |
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