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1、重金屬暴露可以改變秀麗線蟲(chóng)的脂肪積累【摘要】目的:研究重金屬暴露對(duì)秀麗線蟲(chóng)脂肪積累的急性中毒效應(yīng),鑒定重金屬暴露秀麗線蟲(chóng)中發(fā)生表達(dá)模式改變的與脂肪酸代謝相關(guān)的基因。方法:銀、鎘、鉻、銅、汞、錳和鋅作為測(cè)試重金屬,NileRed熒光探針染色分析重金屬暴露線蟲(chóng)腸道的脂肪酸含量變化,并測(cè)定重金屬暴露線蟲(chóng)中脂肪酸代謝相關(guān)基因的轉(zhuǎn)錄水平變化。結(jié)果:在所分析重金屬中,高濃度銀、鉻和銅暴露可以顯著誘導(dǎo)線蟲(chóng)腸道脂肪積累的增加,而高濃度鎘暴露可以顯著降低線蟲(chóng)腸道脂肪的積累。而且,與對(duì)照組相比,高濃度的銀和鉻暴露不明顯影響基因pod2、gei7、lbp8、gpd3、odelorgan
2、ismsareapoentalcluescriticaltohumanhealthanddisease.Sofar,ithasbeenproventhattheC.elegansisanexcellentmodeltoanalyzethemechanismsoffatstorage[5].n),andzinc(Zn)etalsbothbecauseoftheavailabilityoftoxicologicaldataintheliteratureregardingnematodesandotherorganismsandbecauseoftheirabundancein
3、theenvironment.Theaimsofthepresentstudyetalexposureonfatstorageofnematodes,andtoidentifythesensitivegenesrequiredforthemetabolismoffattyacidinmetalexposednematodes. 1Materialsandmethods 1.1Chemicals Themetalconcentrationsusedinthisstudyol·L-1,respectively.AllthechemicalsSigmaAldrich(S
4、t.Louis,MO,USA). 1.2Strains AllnematodesusedtheCaenorhabditisGeicsCenter(CGC).Theyaintainedonnematodegroedium(NGM)platesseededatodesixture(0.45mol·L-1NaOH,2%HOCl).AgesynchronouspopulationsofN2(L4larvaestage)atodesodifiedKmediumonce(50mmol·L-1NaCl,30mmol·L-1KCl,10mmol·L-1NaOAc,pH5.5)[16
5、].Exposuresedin12ethodedasdescribed[17].NileRed(5Hbenzo[α]phenoxazine5one,9diethylamino)pol-1,dilutedinphosphatebufferedsaline(PBS)andaddedontopofNGMplatesalreadyseededl-1.Nematodesinechannel).TheirstainingphenotypesonitoredandevaluatedusingImageJSoftage)bydeterminingaverageintensity
6、ineachanimalsintestine.Morethan30nematodesetalexposednematodes Poly(A)+RNAofnematodesatodespellets1literofmixedstageliquidcultures.Atotalof1μgofpoly(A)+RNAperlaneol·L-1formaldehydeand0.02mol·L-1MOPS,andtransferredontoanylonmembranebyamildalkalinetransfermethod.Northernhybridizationsoftr
7、ansferredpoly(A)+RNAedusingPCRproductprobeslabeledersinetheequalloadingforeachsample,andprimersspecificforact1plificationofallDNAfragmentsforRTPCRedfor30cyclesinaPerkinElmer480thermalcycler(PerkinElmer,,USA)usinga55℃annealingtemperatureanda1minextension. 1.5St