The gut microbial-derived metabolites of ellagitannins and ellagic acid,urolithins(Uros)are well-established anti-cancer metabolites according to preclinical studies.However,their efficacy is limited in systemic tissu...The gut microbial-derived metabolites of ellagitannins and ellagic acid,urolithins(Uros)are well-established anti-cancer metabolites according to preclinical studies.However,their efficacy is limited in systemic tissues,including the brain,by phase-II metabolism.Exosomes(EXOs),extracellular vesicles involved in cell signaling with the ability to cross the blood-brain barrier(BBB),could protect polyphenols from metabolism.Therefore,we loaded milk EXOs with Uro-A,Uro-B or IsoUro-A to evaluate their brain delivery and anticancer effects compared to non-encapsulated Uros.In Sprague-Dawley rats,perfused brain tissue analyses by UPLC-ESI-QTOF-MS showed higher Uro levels(~3-4-fold)at 5 min following intravenous administration of EXO-Uros compared to non-encapsulated Uros,except for Uro-B,using similar Uro concentrations(17-30μM).Experiments on neuro-blastoma SH-SY5Y cells revealed dose-dependent antiproliferative effects for all EXO-Uros(0.3-1.2μM),but not with non-encapsulated Uros(10μM).Flow cytometry analyses indicated that only EXO-Uros arrested the cell cycle and induced apoptosis.Finally,using fluorescent-labeled EXOs and selective inhibitors,the primary endocytic pathway was revealed to be clathrin-dependent.Overall,encapsulating Uros into EXOs is promising for enhancing brain delivery and anticancer activity.展开更多
基金supported by the by the grants TED 2021-130962B-C21 and TED 2021-130962B-C22 funded byMICIU/AEI/10.13039/501100011033 and by the“European Union"or by the"Eu-ropean Union NextGenerationEU/PRTR",PID 2022-136419OB-100,and PID 2022-1431000B-I00funded byMICIU/AEI/10.13039/501100011033 and"ERDF A way of making Europe",the grant 22030/PI/22 funded by the Programa Regional de Fomento de la Investigación Cientifica y Técnica(Plan de Actuación 2022)de la Fundación SénecaAgencia de Cienciay Tecnología de la Región de Murcia(Murcia,Spain),and the AGROALNEXT program supported by MCIN with funding from the European Union NextGenerationEU(PRTR-C17.I1)and Fundación Séneca.M.C.L.-H.is a recipient of a Juan de la Cierva Grant IJC 2020-044353-/MCIN/AEI/10.13039/501100011033/EU/PRTR.A.S.-L.is supported by a predoctoral fellowship(N°2021-01-PhD GRANT)from the International Olive Council.CIBEROBN(CB22/03/00068)is an initiative of the Instituto de Salud Carlos Ⅲ,Spain.
文摘The gut microbial-derived metabolites of ellagitannins and ellagic acid,urolithins(Uros)are well-established anti-cancer metabolites according to preclinical studies.However,their efficacy is limited in systemic tissues,including the brain,by phase-II metabolism.Exosomes(EXOs),extracellular vesicles involved in cell signaling with the ability to cross the blood-brain barrier(BBB),could protect polyphenols from metabolism.Therefore,we loaded milk EXOs with Uro-A,Uro-B or IsoUro-A to evaluate their brain delivery and anticancer effects compared to non-encapsulated Uros.In Sprague-Dawley rats,perfused brain tissue analyses by UPLC-ESI-QTOF-MS showed higher Uro levels(~3-4-fold)at 5 min following intravenous administration of EXO-Uros compared to non-encapsulated Uros,except for Uro-B,using similar Uro concentrations(17-30μM).Experiments on neuro-blastoma SH-SY5Y cells revealed dose-dependent antiproliferative effects for all EXO-Uros(0.3-1.2μM),but not with non-encapsulated Uros(10μM).Flow cytometry analyses indicated that only EXO-Uros arrested the cell cycle and induced apoptosis.Finally,using fluorescent-labeled EXOs and selective inhibitors,the primary endocytic pathway was revealed to be clathrin-dependent.Overall,encapsulating Uros into EXOs is promising for enhancing brain delivery and anticancer activity.
