ghrelin对体外牛卵丘细胞增殖和卵母细胞成熟影响的分析-effect of ghrelin on proliferation and oocyte maturation of bovine cumulus cells in vitro.docx

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2018-06-04 发布于上海
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ghrelin对体外牛卵丘细胞增殖和卵母细胞成熟影响的分析-effect of ghrelin on proliferation and oocyte maturation of bovine cumulus cells in vitro.docx

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ghrelin对体外牛卵丘细胞增殖和卵母细胞成熟影响的分析-effect of ghrelin on proliferation and oocyte maturation of bovine cumulus cells in vitro

Effects ofGhrelinonBovineCumulus ProliferationandOocyte MaturationinvitroAbstractGhrelinistheorexigenicguthormonethathasbeenputforwardtoregulatethe balancebetweenenergyandreproductioninmammals,andistheendogenousligand ofgrowthhormonesecretionreceptor(GHS-R),distributioninendocrinecellsof gastrictissue.Inrecentyears,Ghrelinhasbeendemonstratedthatitplaysessential rolesinreproductivesystembecauseofthehighexpressionofGhrelininthe reproductivegonads.Oocytematurationanddevelopmentisaprerequisitefor fertilizationandthesubsequentembryodevelopmentinmammals,however,the processofoocytematurationisaffectedbymultiplefactors.Ghrelinisanewtypeof hormoneaffectedthereproductivesystem.Therefore,ithasanimportanttheoretical andpracticalsignificanceforpromotingfertilizationinvitroandembryoengineering technologydevelopmentandapplicationtoexplorethepotentialroleofghrelinin oocytematurationprocess.IthasbeendemonstratedthatGhrelinandGHS-Rmainlyexpressedintheluteal cellsandinterstitialcellswithovariansteroidsecretionactivity,andGhrelinand ghrelinreceptorexpressedinoocyteaswell.Moreover,Ghrelinstimulatedgranulosa cellproliferationandinhibitedapoptosisinvitro,anddelayedoocytematuration. WhetherGhrelinwouldberegulatedbovineoocytematurationandcumuluscell proliferationhasnotyetbeenreported.Inthisstudy,thetechnologiesof immunofluorescencestaining,real-timePCR,proliferationassayandWestern BlottingwereadoptedtoresearchtheexpressionofGhrelinandGHS-R1ainbovineoocytesandcumuluscellandtheeffectsofGhrelinand[D-Lys3]-GHRP-6onbovineoocytematurationandcumulus cellproliferationinvitro.The results areas follows:Ithasbeenconfirmedthat Ghrelinproteinexpressedinthecytoplasmof bovineoocytes,GHS-Rproteinspecificallylocatedintheoocytecellsurfaceand GhrelinandGHS-R proteinalsoexpressedincumuluscell.Inthecourseofmaturationinvitro,GhrelinandGHS-R1amRNArelative expressionlevelinoocytedecreased significantlywithmatureprocess;ghrelinmRNA expressionlevelsincumuluscellsshowedprominentlyincreasedat8handdecreased at16h,while