二氧化硅刺激A549细胞水通道蛋白-1表达：体外研究

Abstract:［Objective］Tostudytheexpressionandsignificanceofaquaporin-1(AQP-1)inhumanalveolarepitheliallineA549cellsstimulatedbysilica.［Methods］A549cellsweredividedintocontrol,silica-stimulated,andinhibitorgroups.Thecellsofthesilica-stimulatedgroupandtheinhibitorgroupwerestimulatedby50mg/LsilicondioxidedispersedsuspensiontodetecttheirmRNAexpressionsafter0.5,1,2,4,and8handproteinexpressionsafter3,6,12,and24h.Thecellsoftheinhibitorgroupwerepretreatedwithmercuricchloride(specificchannelinhibitorofAQP-1)for3minutesandthenstimulatedbysilicondioxidefor2htodetectmRNAexpressionorfor6htodetectproteinexpression.TheexpressionofAQP-1proteinwasdetectedbyWesternblotandimmunocytochemistryandtheexpressionofAQP-1mRNAofthecellswasdetectedbyreal-timepolymerasechainreaction.［Results］Comparedwiththecontrolgroup,theexpressionlevelofAQP-1mRNAinsilica-stimulatedcellswas2.78,3.52,3.85,and1.98timesafter0.5,1,2,and4hofsilica-stimulationrespectively(P<0.01),andreturnedtothecontrollevelafter8h;theexpressionleveloftheinhibitorgroupwas65%ofthecorrespondingsilica-stimulatedgroup.AccordingtoWesternblotassay,theexpressionlevelofAQP-1proteinofthesilica-stimulatedcellswas1.44,2.56,1.93,and1.35timesofthecontrolgroupafter3,6,12,and24hofsilica-stimulationrespectively(P<0.05orP<0.01);theleveloftheinhibitorgroupwas70%ofthecorrespondingsilica-stimulatedgroup(P<0.01).Accordingtoimmunocytochemistry,theexpressionlevelofAQP-1proteinwas1.17,1.49,1.29,and1.07timesofthecontrolgroupaftersilicaadministrationfor3,6,12,and24hrespectively(P<0.05orP<0.01);thatoftheinhibitorgroupwas71%ofthecorrespondingsilica-stimulatedgroup(P<0.01).［Conclusion］ElevatedexpressionlevelsofAQP-1mRNAandproteinareinducedbysilica,andthespecificinhibitormercuricchloridecouldinhibittheincreasedexpression,suggestingthatAQP-1participatesthedevelopmentofsilicosis.