Role of the Endogenous Antioxidant System in the Protection of Schistosoma mansoni Primary Sporocysts against Exogenous Oxidative Stress 英文参考文献.docVIP

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Role of the Endogenous Antioxidant System in the Protection of Schistosoma mansoni Primary Sporocysts against Exogenous Oxidative Stress 英文参考文献.doc

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Role of the Endogenous Antioxidant System in the Protection of Schistosoma mansoni Primary Sporocysts against Exogenous Oxidative Stress 英文参考文献

RoleoftheEndogenousAntioxidantSysteminthe ProtectionofSchistosomamansoniPrimarySporocysts againstExogenousOxidativeStress MarinadeMoraesMoura?o1.,NathalieDinguirard2.,Glo′riaR.Franco1,TimothyP.Yoshino2* 1Departamento de Bioqu?′mica e Imunologia, Instituto de Cie?ncias Biolo′gicas, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brasil, 2Department of PathobiologicalSciences,UniversityofWisconsin,Madison,Wisconsin,UnitedStatesofAmerica Abstract Antioxidants produced by the parasite Schistosoma mansoni are believed to be involved in the maintenance of cellular redoxbalance,thuscontributingtolarvalsurvivalintheirintermediatesnailhost,Biomphalariaglabrata.Here,wefocused on specific antioxidant enzymes, including glutathione-S-transferases 26 and 28 (GST26 and 28), glutathione peroxidase (GPx),peroxiredoxin1and2(Prx1and2)andCu/Znsuperoxidedismutase(SOD),knowntobeinvolvedincellularredox reactions,inanattempttoevaluatetheirendogenousantioxidantfunctionintheearly-developingprimarysporocyststage ofS.mansoni.PreviouslywedemonstratedaspecificandconsistentRNAinterference(RNAi)-mediatedknockdownofGST26 and28,Prx1and2,andGPxtranscripts,andanunexpectedelevationofSODtranscriptsinsporocyststreatedwithgene- specificdouble-stranded(ds)RNA.Inthepresentfollowupstudy,invitrotransformingsporocystswereexposedtodsRNAs forGST26and28,combinedPrx1/2,GPx,SODorgreen-fluorescentprotein(GFP,control)for7daysinculture,followedby assessment of the effects of specific dsRNA treatments on protein levels using semi-quantitative Western blot analysis (GST26,Prx1/2only),andlarvalsusceptibilitytoexogenousoxidativestressininvitrokillingassays.Significantdecreases (80%and50%)inimmunoreactiveGST26andPrx1/2,respectively,wereobservedinsporocyststreatedwithspecificdsRNA, comparedtocontrollarvaetreatedwithGFPdsRNA.SporocystsculturedwithdsRNAsforGST26,GST28,Prx1/2andGPx, butnotSODdsRNA,weresignificantlyincreasedintheirsusceptibilitytoH O oxidativestress(60–80%mortalitiesat48hr) 2 2 compared to GFP