简介:目的:探讨胎牛血清对小鼠角膜上皮细胞生长和分化的影响。方法:分别在无血清低钙培养基(KSFM)和含100mL/L胎牛血清的KSFM中培养小鼠角膜上皮细胞。比较两种培养基中角膜上皮细胞的群体倍增(PD)。通过RT-PCR和Westernblotting方法检测p63、角蛋白19以及involucrin的表达。结果:在KSFM中,小鼠角膜上皮细胞可稳定传代超过25代,但在含胎牛血清的培养基中,细胞仅能存活3代。在KSFM中,培养细胞呈典型铺路石样外观,RT-PCR和Westernblotting结果显示细胞表达p63、角蛋白19以及involucrin。当培养基中加入胎牛血清后,细胞形态明显增大,呈扁平状;involucrin表达显著增强。结论:胎牛血清可抑制小鼠角膜上皮细胞增生、诱导其分化。
简介:AIM:ToInvestigatetheeffectsoftransforminggrowthfactorβ2(TGF-β2)andconnectivetissuegrowthfactor(CTGF)ontransdifferentiationofhumanlensepithelialcells(HLECs)culturedinvitroandsynthesisofextracellularmatrix(ECM).METHODS:HLECsweretreatedwithTGF-β2(0,0.5,1.0,5,10μg/L)andCTGF(0,15,30,60,100μg/L)fordifferenttimes(0,24,48,72h)invitroandtheexpressionofα-smoothmuscleactin(α-SMA),themaincomponentoftheextracellularmatrixtypeⅠcollagen(Col-1)andfibronectin(Fn)weremeasuredbyusingreal-timepolymerasechainreaction(PCR)andwestern-blot.RESULTS:TGF-β2andCTGFsignificantlyincreasedexpressionofα-SMAmRNAandprotein(P<0.05,P<0.001),FnmRNAandprotein(P<0.001),Col-1mRNAandprotein(P<0.001).TGF-β2couldinduceHLECsexpressionofCTGFmRNAandproteinindosedependentmanner(P<0.05,P<0.001).TGF-β2andCTGFcouldinduceHLECstoexpressα-SMA,FnandCol-1intime-dependentmanner.EachtimeofTGF-β2andCTGFinducedHELCsexpressionofα-SMA,Fn,Col-1mRNAandproteinwassignificantincreasecomparedwithcontrol(P<0.05,P<0.001).CONCLUSION:TGF-β2andCTGFcouldinduceHLECsepithelialmesenchymaltransitionandECMsynthesis.
简介:AIM:ToinvestigatethesideeffectsofthecommonlyusedlasertreatmentalongwithtestingtheneuroprotectiveeffectofbFGFonapotentialretinalimpairment.METHODS:Todothis,30chinchillapigmentedadultmalerabbitsweredividedintothecontrolandexperimentalgroups.ThecontrolandexperimentalgroupsunderwentbothlaserapplicationandbFGFtreatment.Theretinaltissueimpairmentanditsrenewalrateweretestedunderthelightandelectronmicroscopicallevels.RESULTS:Thefocallaserapplicationonrabbiteyescausedmorphologicalalterationsbothintheapplicationregionandintheneighbouringareas.Inthedamagedareas,theouternuclearlayeroftheneuralretinawasalmostdisappeared,retinapigmentepitheliumwasinterrupted,theretinapigmentepitheliummigratedintraretinally,andthedamagedregionalongwithneighbouringareasseemedtobenotseparated.bFGFapplicationjustafterthelaserphotocoagulation,revealedbetterresultsinapplicationareas.CONCLUSION:ItcouldbesuggestedthatthebFGFapplicationfollowinglaserphotocoagulationmighthaveprotective,repairingandwoundhealingeffectsontheretina.
简介:Thekeratoprosthesis(KPro;artificialcornea)isaspecialrefractivedevicetoreplacehumancorneabyusingheterogeneousformingmaterialsfortheimplantationintothedamagedeyesinordertoobtainacertainvision.Themainproblemsofartificialcorneaarethebiocompatibilityandstabilityofthetissueparticularlyinpenetratingkeratoplasty.Thecurrentstudiesoftissue-engineeredscaffoldmaterialsthroughcomprisingcompositesofnaturalandsyntheticbiopolymerstogetherhavedevelopedanewwaytoartificialcornea.Althoughawideagreementthatthelong-termstabilityofthesedeviceswouldbegreatlyimprovedbythepresenceofcorneacells,modificationofkeratoprosthesistosupportcorneacellsremainselusive.Mostofthestudiesoncornealsubstratematerialsandsurfacemodificationofcompositeshavetriedtoimprovethegrowthandbiocompatibilityofcorneacellswhichcannotonlyreducethestimulusofheterogeneousmaterials,butalsomoreimportantlycontinuousandstablecorneacellscanpreventthedestructionofcollagenase.Thenecrosisofstromaandspontaneousextrusionofthedevice,allowformaintenanceofaprecornealtearlayer,andplaytheroleofensuringagoodopticalsurfaceandresistingbacterialinfection.Asaresult,improvementincornealcellshasbeenthemainaimofseveralrecentinvestigations;someefforthasfocusedonbiomaterialforitswellbiologicalpropertiessuchaspromotingthegrowthofcorneacells.Thepurposeofthisreviewistosummarythegrowthstatusofthecornealcellsaftertheimplantationofseveralartificialcorneas.
简介:目的:评价聚乙二醇滴眼液联合重组牛碱性成纤维细胞生长因子滴眼液治疗干眼症的疗效。方法:干眼症患者51例102眼,双眼自身对照,随机分为治疗组和对照组。治疗组给予聚乙二醇滴眼液和重组牛碱性成纤维细胞生长因子滴眼液,均4次/d,两种滴眼液之间间隔5~10min。对照组给予聚乙二醇滴眼液滴眼,4次/d,连续用药1mo后复查。观察每组用药前后SchirmerI试验,BUT,角膜荧光素染色和症状改善情况。结果:两组治疗前后BUT,角膜荧光素染色和症状都有显著差异(P〈0.05)。两组治疗前后SchirmerI试验结果无显著差异(P〉0.05)。治疗后两组BUT,角膜荧光素染色和症状也有显著差异(P〈0.05)。结论:聚乙二醇滴眼液联合重组牛碱性成纤维细胞生长因子滴眼液对治疗干眼症有明显的疗效。
简介:AIM:Toexaminetheexpressionofsurvivinandvascularendothelialgrowthfactor(VEGF)duringthedevelopmentofretinalneovascularization(NV)inamousemodel.·METHODS:Awell-characterizedmurinemodelofretinalNVwasusedtostudytheexpressionofsurvivinandVEGF.NVoftheretinawasinducedinmicebyexposureto75%O2frompostnataldayP7toP12,followedbyreturntoroomairfromP12toP17.ExpressionofsurvivinandVEGFproteinwasanalyzedbyImmunohistochemistry.Inaddition,mousemodelofproliferativeretinopathywasanalyzedbyretinalfluoresceinangiographyandquantificationanalysis.·RESULTS:Thenormalmicehadbothsuperfiekalanddeepvascularlayersthatextendedfromtheopticnervetotheperiphery.Inintraocularpressure(IOP)micewerecharacterizedbyrepresentatypicalpatternofpathologicalretinalNV.Therearelessorlittlenucleiofnewvesselsvascularendothelialcellbreakingthroughtheinnerretinalthaninretinopathyofprematurity(ROP)mice,largeclustersofbloodvesselswereadherenttotheinternallimitingmembrane(ILM)(0.27±0.20vs23.38±1.027,t=9.454,P<0.001).DuringtheangiogenicperiodfromP13toP17,survivinandVEGFproteinexpressionincreasedinexperimentalretinascomparedwithcontrolsamples(2.56±0.46vs3.34±0.40,t=17.43,P<0.01:2.18±0.75vs4.34±0.25,t=19.61,P<0.01).ProteinlevelsofVEGFandsurvivnhassignificantlypositivecorrelation(P<0.05,r=0.411).·CONCLUSION:CorrelationwasmadeattheproteinlevelsofsurvivinexpressioncomparedwiththatofVEGFinamurinemodelofretinalNV,whichsuggestsatemporalroleforsurvivinandVEGFinnewvesselformationinresponsetohypoxicstimulation.
简介:目的:研究增殖细胞核抗原(proliferatingcellnuclearanti-gen,PCNA)和胰岛素样生长因子Ⅱ(insulin-likegrowthfactor-Ⅱ,IGF-Ⅱ)在翼状胬肉中表达的关系。方法:应用免疫组织化学SABC法检测40例原发性翼状胬肉组织标本,10例正常结膜组织标本中PCNA和IGF-Ⅱ蛋白的表达情况。结果:翼状胬肉组中PCNA,IGF-Ⅱ表达均高于正常结膜组(P〈0.01),IGF-Ⅱ与PCNA蛋白表达之间的相关系数为0.731(P〈0.01)。结论:翼状胬肉为一种具有肿瘤潜能的增生性眼表疾病。IGF-Ⅱ通过促进细胞增殖参与了翼状胬肉的发生、发展。
简介:AIM:TodiscusstheimpactofLyciumBarbarumPolysaccharide(LBP)andDanshensupurifiedfromTraditionalChineseMedicine(TCM)onvascularendothelialgrowthfactor(VEGF)ofrabbitswithretinalneovascularization.METHODS:Fortyrabbitsweredividedintonormalcontrolgroup,modelcontrolgroup,LBPgroupandDanshensugroup.Animalsinthenormalcontrolgroupwerefedinthenormaloxygenenvironment.Animalsintheotherthreegroupswereputintotheenvironmentwith70%oxygenfor5daysinordertobuildthemodelofoxygen-inducedvascularproliferationretinopathy.AndthendifferentTCMextractwasinjectedintotheabdominalcavitiesoftheseannimals.After7days,theVEGFcontentofintheserumofrabbitwasmeasuredbydoubleantibodysandwichmethod.RESULTS:DataanalysisindicatedthatVEGFcontentwasasfollows:Danshensugroupwaslowerthanmodelcontrolgroup(12.92±3.84ng/Lvs19.32±4.15ng/L,P<0.05);LBPgroupandnormalcontrolgroupwerelowerthanmodelcontrolgroup(12.92±3.84ng/L,9.26±1.61ng/Lvs19.32±4.15ng/L,P<0.01);totalbloodviscosity,plasmaviscosity,cholesterolcontent,fibrinogencontentandtriacylglycerolcontentafterperitonealinjectionofLBPandDanshensuwereobviouslylowerthanbeforeinjection.CONCLUSION:TCMextract-LBPandDanshensucanprominentlyreducethecontentofVEGFintheprocessofvascularproliferativeretinopathyofrabbit;canpreventtheoccurrenceofretinalmicrovasculardiseasebyimprovingpartialoxygen-deficientenvironmentoraffectingallkindsofnewgrowthfactor.
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.