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1、摘要综合以上研究结果，我们认为miR-224正是通过靶向于不同的靶基因实现TGF-β1对卵泡发育调节的介导作用。而对miRNA参与卵泡发育过程的进一步研究将会为人工干预生殖过程提供研究基础，如治疗生殖紊乱等。关键词：microRNA-224小鼠卵泡发育细胞增殖COC扩展IIAbstractABSTRACTManymembersoftheTGF-βsuperfamilyareindicatedtoplayimportantrolesinmammalianreproduction,suchasaffectingtheearlyfolliculogenesis,granulosac
2、ellproliferationanddifferentiation,oocytematurationandembryoimplantationandthemaintanenceofpregnancy.TheperturbationoftheTGF-βsignalingtransductioncouldresultinfemaleinfertility.MicroRNAs(miRNAs),assmallnon-codingRNAs,wererecentlyfoundtoregulategeneexpressionatthepost-transcriptionallevels,
3、eitherinhibitorpromote.However,littleisknownabouttheroleofmiRNAsinTGF-β-mediatedgranulosacellproliferation，hormonereleaseandCOCexpansion.Inthisstudy,themiRNAexpressionprofilingwasidentifiedfromTGF-β1-stimulatedmousepreantralgranulosacells(GCs),and3miRNAswerefoundtobesignificantlyup-regulate
4、dand13miRNAsweresignificantlydown-regulated.Amongtheup-regulatedmiRNAs,miR-224wasthesecondmostsignificantlyelevatedmiRNA.Thisup-regulationwasattenuatedbytreatmentofGCswithSB431542(aninhibitorofTGFβsuperfamilytypeIreceptors,thusblockingthephosphorylationofthedownstreameffectorsSmad2/3),indic
5、atingthatmiR-224expressionwasregulatedbythecanonicalTGF-β1/Smadspathway.Byaseriesstudiesofluciferaseanalysisandwesternblot,weidendifiedthatsmad4isoneofthetargetofmiR-224.TheectopicexpressionofmiR-224bytransfectingwithmiR-224mimicscanenhanceTGF-β1-inducedGCproliferationthroughtargetingsmad4.
6、InhibitionofendogenousmiR-224partiallysuppressedGCproliferationinducedbyTGF-β1.Inaddition,bothmiR-224andTGF-β1canpromoteestradiolrelease,oneofthemostsignificantrolesofGCinreproduction.Byusingthereal-timePCRanalysis,weproposethatthefunctionsofestradiolreleasemaypartlythroughincreasingthecyp1
7、9a1mRNAlevels.ThisisthefirstdemonstrationthatmiRNAscancontrolreproductivefunctionsbypromotingTGF-β1-inducedGCproliferationandovarianestrogenrelease.ByanalyzingthemRNAexpressionprofileinGCstreatedbyTGF-β1,wefoundtheremanygeneswerede-regulated,ofwhich463mR
