简介：AIM:ToinvestigatetheeffectofY-27632onthesurvivalandneuriteoutgrowthoftheculturedretinalneurocytes.METHODS:Afterthepostnatalday2-3,Sprague-Dawleyretinalneurocyteswereculturedfor48hours,theculturemediawasreplacedwithserum-freemedia(controlgroup)andserum-freemediacontained30μmol/LY-27632(Y-27632group),andthecellswerecontinuallyculturedanother48hours.Theculturedretinalneurocyteswereidentifiedwithanti-neuronspecificenolase(NSE)immunocytochemistry.ThesurvivalstateofthosecellswasestimatedbyMTTassay,andtheneuriteoutgrowthofthosecellswasevaluatedbythecomputerizedimage-analysissystem.RESULTS:Comparedwiththecontrolgroup,theabsorbancevaluesofcellssurvivalinY-27632groupincreased12.90%and33.33%respectivelyafter72and96hoursculture.Y-27632hadnosignificanteffectonthediameterofculturedretinalneurocytes.Comparedwiththecontrolgroup,Y-27632inducedastableimprovementofneuriteoutgrowthofretinalneurocytesafter72and96hoursculture(P=0.001).CONCLUSION:Y-27632couldpromotethesurvivalandneuriteoutgrowthoftheearlypostnatalculturedretinalneurocytes.

简介：AIM:ToinvestigatetheinterferingeffectofY-27632,aROCK-Iselectiveinhibitor,onthesignaltransductionpathwayoftransforminggrowthfactor-β1(TGF-β1)inocularTenoncapsulefibroblasts(OTFS)invitro.METHODS:AfterOTFSfrompassages4to6invitrowereinducedbyTGF-β1andthentreatedbyY-27632,thechangesoftheOTFScellcycleswereanalyzedviaflowcytometry,andtheproteinsexpressionoftheα-smoothmuscularactin(α-SMA),connectivetissuegrowthfactor(CTGF),collagenIwerecalculatedbyWesternblot.AfterOTFStreatedbythedifferentconcentrationsofY-27632,theexpressionlevelsoftheα-SMA,CTGFandcollagenImRNAwereassayedbyRT-PCR.RESULTS:Y-27632hadnomarkedlyeffectontheOTFScellcycles.AftertreatedbyTGF-β1,OTFSinG1periodsignificantlyincreased.ThecellcyclesdistributionbybothTGF-β1andY-27632hadnoremarkabledifferencefromthatincontrolgroup.Y-27632significantlyinhibitedtheproteinsexpressionsofbothα-SMAandCTGF,whiletosomeextentinhibitedthatofcollagenI.TGF-β1significantlypromotedtheproteinsexpressionsofα-SMA,CTGFandcollagenI.AfterOTFStreatedbybothTGF-β1andY-27632,ofα-SMA,theproteinexpressionwassimilarwiththatincontrolgroup(P=0.066>0.05),buttheproteinexpressionofCTGForcollagenI,respectively,wassignificantlydifferentfromthatincontrolgroup(P=0.000<0.01).Thedifferencesofexpressionsoftheα-SMA,CTGFandcollagenImRNAin30,150,750μmol/LY-27632groupwerestatisticallysignificant,comparedwiththoseincontrolgroup,respectively(α-SMA,P=0.002,0.000,0.000;CTGF,P=0.014,0.002,0.001;collagenI,P=0.003,0.002,0.000).CONCLUSION:BlockingtheRho/ROCKsignalingpathwaybyusingofY-27632couldinhibitthecellularproliferationandtheexpressionofbothCTGFandα-SMAwhateverOTFSinducedbyTGF-β1ornot.Y-27632suppressedtheexpressionofcollagenImRNAwithoutinduction.