简介：AIM:Toinvestigatewhether15-Lipoxygenase-1(15-LOX-1)playsanimportantroleintheregulationofangiogenesis,inhibitinghypoxia-inducedproliferationofretinalmicrovascularendothelialcells(RMVECs)andtheunderlyingmechanism.METHODS:PrimaryRMVECswereisolatedfromtheretinasofC57/BL6JmiceandidentifiedbyanevaluationforFITC-markedCD31.ThehypoxiamodelswereestablishedwiththeBio-bagandevaluatedwithablood-gasanalyzer.ExperimentswereperformedusingRMVECstreatedwithandwithouttransferAd-15-LOX-1orAd-vectorbothunderhypoxiaandnormoxiaconditionat12,24,48,72hours.Theefficacyofthegenetransferwasassessedbyimmunofluorescencestaining.CellsproliferationwasevaluatedbytheCCK-8method.RNAandproteinexpressionsof15-LOX-1,VEGF-A,VEGFR-2,eNOsandPPAR-rwereanalyzedbyreal-timereversetranscriptionpolymerasechainreaction(RT-PCR)andWesternblot.RESULTS:RoutineevaluationforFITC-markedCD31showedthatcellswerepure.Theresultsofblood-gasanalysisshowedthatwhenthecultureswereexposedtohypoxiaformorethan2hours,thePo2was4.5to5.4Kpa.WeverifiedRMVECscouldbeinfectedwithAd-15-LOX-1orAd-vectorviaFluorescencemicroscopy.CCK-8analysisrevealedthattheproliferativecapacitiesofRMVECsinhypoxicgroupweresignificantlyhigherateachtimepointthantheywereinnormoxicgroup(P<0.05).Inahypoxiccondition,theproliferativecapacitiesofRMVECsin15-LOX-1groupweresignificantlyinhibited(P<0.05).Real-timeRT-PCRanalysisrevealedthattheexpressionsofVEGF-A,VEGF-R2andeNOsmRNAincreasedinhypoxiagroupcomparedwithnormoxiagroup(P<0.01).However,theexpressionsof15-LOX-1,PPAR-rmRNAdecreasedinhypoxiagroupcomparedwithnormoxiagroup(P<0.01).Italsoshowedthatinahypoxiccondition,theexpressionsofVEGF-A,VEGF-R2andeNOsmRNAdecreasedsignificantlyin15-LOX-1groupcomparedwithhypoxiagroup(P<0.01).However,15-LOX-1andPPAR-rmRNAincreasedsigni

简介：AIM:ToinvestigatetheeffectofintravitrealinjectionofDL-alpha-aminoadipicacid(DL-α-AAA)onocularrefractivestateandretinaldopamine,transforminggrowthfactor-β2(TGFβ2),vasoactiveintestinalpolypeptide(VIP)inguineapigform-deprivedmyopia.METHODS:Four-week-oldpigmentedguineapigswererandomlyassignedto4groups:normalcontrol,deprivation,deprivationplusDL-α-AAA,deprivationplussaline.Formdeprivationwasinducedwiththeself-madetranslucenteyeshields,andlastedfor14days.8μgDL-α-AAAwasinjectedintothevitreouschamberofdeprivedeyes.Thecornealradiusofcurvature,refractionandaxiallengthweremeasured.Retinaldopaminecontentwasevaluatedbythehigh-performanceliquidchromatographywithelectrochemicaldetection,andTGFβ2andVIPproteinweredetectedbyWesternblotting.RESULTS:Fourteendaysofeyeocclusioncausedtheaxiallengthtoelongateandbecomemyopicintheform-deprivedeyes,withthedecreaseofretinaldopamineandtheincreaseofTGFβ2andvasoactiveintestinalpolypeptide(VIP)protein.IntravitrealinjectionofDL-α-AAAcouldinhibitthemyopicshiftfrom(-3.65±1.06)Dto(-1.48±0.63)D,P<0.01duetogogglesoccludingandcausethedecreaseofretinalTGFβ2proteininthedeprivedeyes.However,intravitrealinjectionofDL-α-AAAhadnosignificanteffectonretinaldopamineandVIPproteinindeprivedeyes.RetinalTGFβ2proteincorrelatedhighlywiththeocularrefraction(y=-3.34+0.31/x,F=74.75,P<0.001)andaxiallength(y=8.39-0.02/x,F=48.32,P<0.001)indifferenttreatmentgroups.·CONCLUSION:IntravitrealinjectionofDL-α-AAAiseffectivelyabletosuppressthedevelopmentofformdeprivationmyopia,whichmaybeassociatedwithretinalTGFβ2proteininguineapigs.

简介：AIM:ToinvestigatetheexpressionsoftypeIcollagen,α2integrinandβ1integrinintheposteriorscleraofguineapigswithdefocusmyopiaandwhetherbasicfibroblastgrowthfactor(bFGF)injectioninhibitstheformationanddevelopmentofmyopiabyupregulatingtheexpressionoftypeIcollagen,α2integrinandβ1integrin.METHODS:After14daysoftreatment,therefractivestateandaxiallengthweremeasuredandthelevelsoftypeIcollagen,α2integrinandβ1integrinwereassayedintheposteriorscleraeofgroupsofguineapigsthatworeamonocular-7Dpolymethylmethacrylate(PMMA)lensorhad-7DlenswearfollowedbytheperibulbarinjectionofPhosphateBufferSolution(PBS)orbFGF.Theuntreatedfelloweyeservedasacontrol.Guineapigswithnotreatmentservedasnormalgroup.·RESULTS:Theresultsshowedthat14daysofmonoculardefocusincreasedaxialeyelengthandrefraction,whilebFGFdeliveryinhibitedthemmarkedly.Further,itwasalsofoundthatthemonocular-7DlenscoulddecreasethelevelsoftypeIcollagen,α2integrinandβ1integrinexpressions,while,unlikePBS,bFGFincreasedthemsignificantlyincomparisontocontralateralcontroleyesandnormaleyes.CONCLUSION:bFGFcanpreventtheformationanddevelopmentofdefocusmyopiabyupregulatingtheexpressionsoftypeIcollagen,α2integrinandβ1integrin.Takentogether,ourresultsdemonstratethatbFGFpromotesscleraremodelingtopreventmyopiainguineapigs.