Some mycotoxins such as beauvericin (BEA), ochratoxin A (OTA), and zearalenone (ZEA) can cross the blood brain barrier, which is why we tested the anti-inflammatory action of a pumpkin carotenoid extract (from the pulp) against these mycotoxins and their combinations (OTA+ZEA and OTA+ZEA+BEA) on a blood brain barrier model with co-cultured ECV304 and C6 cells using an untargeted metabolomic approach. The cells were added with mycotoxins at a concentration of 100 nmol/L per mycotoxin and pumpkin carotenoid extract at 500 nmol/L. For control we used only vehicle solvent (cell control) or vehicle solvent with pumpkin extract (extract control). After two hours of exposure, samples were analysed with HPLC-ESI-QTOF-MS. Metabolites were identified against the Metlin database. The proinflammatory arachidonic acid metabolite eoxin (14,15-LTE4) showed lower abundance in ZEA and BEA+OTA+ZEA-treated cultures that also received the pumpkin extract than in cultures that were not treated with the extract. Another marker of inflammation, prostaglandin D2-glycerol ester, was only found in cultures treated with OTA+ZEA and BEA+OTA+ZEA but not in the ones that were also treated with the pumpkin extract. Furthermore, the concentration of the pumpkin extract metabolite dihydromorelloflavone significantly decreased in the presence of mycotoxins. In conclusion, the pumpkin extract showed protective activity against cellular inflammation triggered by mycotoxins thanks to the properties pertinent to flavonoids contained in the pulp.
Pojedini mikotoksini poput bovericina (BEA), okratoksina A (OTA) i zearalenona (ZEA) prelaze krvno-moždanu barijeru, a to je i razlog zbog kojega smo istražili djelovanje ekstrakta karotenoida iz mesa bundeve protiv upalnih procesa izazvanih ovim mikotoksinima i njihovim kombinacijama (OTA+ZEA i OTA+ZEA+BEA) na modelu krvno-moždane barijere koji se sastojao od kultura stanica ECV304 i C6, oslanjajući se pritom na neciljani metabolomički pristup. Stanice su tretirane mikotoksinima u koncentraciji od 100 nmol/L po mikotoksinu odnosno ekstraktom karotenoida u koncentraciji od 500 nmol/L. Za kontrolu smo upotrijebili samo otapalo (stanična kontrola) odnosno otapalo s bundevinim ekstraktom (ekstraktna kontrola). Nakon dva sata tretmana uzorci su analizirani metodom tekućinske kromatografije / masene spektrometrije (HPLC-ESI-QTOF-MS), a dobiveni metaboliti identificirani su usporedbom s bazom podataka Metlin. Primjena ekstrakta značajno je smanjila količinu metabolita proupalne arahidonske kiseline eoksina (14,15-LTE4) u kulturama tretiranima samo zearalenonom odnosno kombinacijom BEA+OTA+ZEA. Drugi upalni biljeg, prostaglandin D2-glicerol ester, otkriven je samo u kulturama tretiranima kombinacijama OTA+ZEA odnosno BEA+OTA+ZEA, ali ne i u onima koje su usto tretirane bundevinim ekstraktom. Osim toga, u prisutnosti mikotoksina značajno je porasla koncentracija metabolita dihidromoreloflavona iz bundevina ekstrakta. Time je taj ekstrakt iskazao zaštitno djelovanje protiv stanične upale uzrokovane mikotoksinima zahvaljujući svojstvima flavonoida koji se nalaze u njezinu mesu.
Keywords: ECV304; beauvericin; bovericin; metabolomics; metabolomika; ochratoxin A; okratoksin A; zearalenon; zearalenone.
© 2021 Manuel Alonso-Garrido, Noelia Pallarés, Guillermina Font, Paola Tedeschi, Lara Manyes, and Manuel Lozano, published by Sciendo.