Rottlerin Stimulates Exosome/Microvesicle Release Via the Increase of Ceramide Levels Mediated by Ampk in an In Vitro Model of Intracellular Lipid Accumulation

Biomedicines

María T. Vallejo¹  Bohdan Babiy²  Óscar Pastor²  Diego Gómez-Coronado³  David García-Seisdedos³  Milagros Lerma³  Yessenia L. Molina³  Javier Martínez-Botas³  Miguel A. Lasunción³  Rebeca Busto³  María J. Casarejos⁴ 

[1] Molecular Imaging and Immunohistochemistry Unit, Hospital Universitario la Paz, IdiPAZ, 28046 Madrid, Spain;Servicio de Bioquímica-Clínica, Hospital Universitario Ramón y Cajal, IRYCIS, 28034 Madrid, Spain;Servicio de Bioquímica-Investigación, Hospital Universitario Ramón y Cajal, IRYCIS, 28034 Madrid, Spain;Servicio de Neurobiología, Hospital Universitario Ramón y Cajal, IRYCIS, 28034 Madrid, Spain;

关键词: rottlerin;    exosome/microvesicle;    intracellular lipid trafficking;    ceramide;    sphingolipid;    lysosomal storage disease;   

DOI  :  10.3390/biomedicines10061316

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown