Differences in the Endocannabinoid System of Sperm from Fertile and Infertile Men 英文参考文献.docVIP

DifferencesintheEndocannabinoidSystemofSperm fromFertileandInfertileMen SheenaE.M.Lewis1.,CinziaRapino2.,MoniaDiTommaso2,MariangelaPucci2,NataliaBattista2, RitaParo1,LukeSimon1,DeborahLutton3,MauroMaccarrone2,4* 1SchoolofMedicine,CentreforPublicHealth,Queen’sUniversityBelfast,InstituteofClinicalScience,Belfast,UnitedKingdom, 2DepartmentofBiomedicalSciences, UniversityofTeramo,Teramo,Italy,3RegionalFertilityCentre,RoyalJubileeMaternityServices,Belfast,UnitedKingdom,4EuropeanCenterforBrainResearch(CERC)/ IRCCSSantaLuciaFoundation,Rome,Italy Abstract Male infertility is a major cause of problems for many couples in conceiving a child. Recently, lifestyle pastimes such as alcohol, tobacco and marijuana have been shown to have further negative effects on male reproduction. The endocannabinoid system (ECS), mainly through the action of anandamide (AEA) and 2-arachidonoylglycerol (2-AG) at cannabinoid(CB1,CB2)andvanilloid(TRPV1)receptors,playsacrucialroleincontrollingfunctionalityofsperm,withaclear impactonmalereproductivepotential.Here,spermfromfertileandinfertilemenwereusedtoinvestigatecontent(through LC-ESI-MS), mRNA (through quantitative RT-PCR), protein (through Western Blotting and ELISA) expression, and functionality (through activity and binding assays) of the main metabolic enzymes of AEA and 2-AG (NAPE-PLD and FAAH,forAEA;DAGLandMAGLfor2-AG),aswellasoftheirbindingreceptorsCB1,CB2andTRPV1.Ourfindingsshowa markedreductionofAEAand2-AGcontentininfertileseminalplasma,paralleledbyincreaseddegradation:biosynthesis ratiosofbothsubstancesinspermfrominfertileversusfertilemen.Inaddition,TRPV1bindingwasdetectedinfertilesperm butwasundetectableininfertile sperm,whereas thatofCB1 andCB2 receptors wasnotstatisticallydifferentinthetwo groups.Inconclusion,thisstudyidentifiedunprecedentedalterationsoftheECSininfertilesperm,thatmightimpacton capacitationandacrosomereaction,andhencefertilizationoutcomes.Thesealterationsmightalsopointtonewbiomarkers todeterminemalerep