EffectofvaspinonhighglucoseandlipidinducedoxidativestressinINS-1cellsviatheNrf2/AREsignalingpathway

Abstract:

Objective：ToinvestigatetheeffectofvaspinonhighglucoseandlipidinducedoxidativestressinINS-1cells.Methods：INS-1cellswereculturedanddividedintonormalcontrolgroup，highglucose（HG）+palmiticacid（PA）group，HG+PA+vaspin（80ng/mL）group，HG+PA+vaspin（160ng/mL）group，HG+PA+vaspin（320ng/mL）group，HG+PA+vaspin（640ng/mL）group.Colorimetryandenzyme-linkedimmunosorbentassaywereusedtomeasureoxidativestress-relatedindicators；meanwhile，DCFH-DAfluorescentprobestaining，glucose-stimulatedinsulinsecretiontest（3.3and16.7mmol/L），flowcytometry，andWesternblotwereusedtodeterminethelevelsofintracellularreactiveoxygenspecies（ROS），insulinsecretion，apoptosis，andproteinexpressionofnuclearfactorerythroid2-relatedfactor2（Nrf2），respectively.Results：ComparedwiththeHG+PAgroup，theHG+PA+vaspin（640ng/mL）grouphadasignificantlyincreasedinsulinsecretionlevelafterstimulationbybothlow-levelandhigh-levelglucose（bothP<0.05）aswellassignificantlyincreasedtotalantioxidantcapacityandlevelsofsuperoxidedismutaseandglutathioneperoxidase（allP<0.05），buthadsignificantlydecreasedlevelsofROS，malondialdehyde，and8-hydroxydeox-yguanosine（allP<0.05）.Withtheincreasingvaspinlevels，thevaspin-treatedgroupshadsignificantlyreducedapoptosisintheINS-1cellsafterinterventionbyhigh-levelglucoseandlipidscomparedwiththeHG+PAgroup，asrevealedbyflowcytometry（allP<0.05），andshowedanincreasingtrendinintracytoplasmicNrf2expressionasrevealedbyWesternblot，whiletheintranuclearNrf2expressionwassignificantlyhigheronlyintheHG+PA+vaspin（640ng/mL）groupcomparedwiththeHG+PAgroup（0.798±0.080vs.0.579±0.065，P=0.039）.Conclusion：VaspincansuppresshighglucoseandpalmiticacidinducedoxidativestressinjuryinINS-1cells，reduceapoptosis，andimproveinsulinsecretionlevelbyactivatingtheNrf2/AREsignalingpathway.