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1、上海第二医科大学学报Vol.20Suppl.2000ActaUniversitatisMedicinalisSecondaeShanghai·5·文章编号:0258-5898(2000)Suppl-0005-03生长因子对大鼠肝细胞增殖和酶活性表达的影响谢青廖丹彭星亮周霞秋(上海第二医科大学瑞金医院传染科,上海200025)摘要:目的研究表皮生长因子(EGF)、肝细胞生长因子(HGF)、EGF联合HGF对大鼠原代培养肝细胞增殖和功能的影响,为肝细胞移植建立细胞库、提高逆转录病毒载体感染肝细胞的效率寻找适当的培养条件。方法分3离、
2、培养肝细胞,采用H-TdR掺入试验测定肝细胞内DNA合成,应用细胞化学方法观察培养的肝细胞内葡萄糖-6-磷酸酶活性的表达。结果培养肝细胞受EGF(10ng/ml)、HGF(5ng/ml)作用后,肝细胞内DNA合成分别增加了59倍(P<0.01)和16倍(P<0.01)。EGF和HGF联合应用不表现出明显的协同作用。培养的肝细胞可合成葡萄糖-6-磷酸酶,具有肝细胞特异性功能。结论提示EGF和HGF在体外可刺激肝细胞增殖并维持肝细胞功能,为肝细胞移植、肝病基因治疗提供了一个完善的肝细胞培养系统。关键词:生长因子细胞增殖酶活性原代培养
3、大鼠肝细胞中图号:Q28R392.4文献标识码:AEffectofGrowthFactorsonProliferationandExpressionofEnzymeActivityofRatHepatocytesinPrimaryCultureXIEQing,LIAODan,PENXingliang,etal.(DepartmentofInfectiousDisease,RuijinHospital,SSMU,Shanghai200025)Abstract:objectiveThestudywasundertakentoeval
4、uatetheeffectofgrowthfactorsonDNAsynthesisandfunctionofadultrathepatocytesmaintainedinprimaryculture,andtoprovideause-ful,welldefinedculturesystemforestablishmentofacellbankandforefficient,stabletransductionofhepatocytesbyreplication-defectiveretroviruses.MethodsThei
5、nsitutwo-stepcollagenaseperfusiontechniquewasusedfortheisolationofhepatocytesfromtheadultliver.Cellsweremaintained3incultureonexposuretoEGF,HGF,andEGFcombinedwithHGF.H-TdRincorporationwasusedtodetermineDNAsynthesis.ResultsTheexpressionofglucose-6-phosphataseactivityw
6、asde-tectedbythecytochemicalprocedure.EGF(10ng/ml)andHGF(5ng/ml)couldstimulatetheDNAsyn-thesisofculturedhepatocytessignificantly.Theaverageincreaseinthecontrolgroupwas59timeswithEGF(P<0.01)and16timeswithHGF(P<0.01)respectively.Nosynergiceffectwasobservedinhepatocytes
7、treatedwithEGFplusHGF.Mostoftheculturedcellsexhibitedglucose-6-phosphataseactivities.ThepresenceofEGFandHGFinrathepatocyteculturescanincreaseDNAsynthesisandmaintainhepatocytefunction.ConclusionItsuggeststhatthehepatocyteculturesdescribedinthepresentstudymaybeauseful,
8、optimalculturesystemforthehepatocytetransplantationandlivergenetherapy.Keywords:growthfactorscellproliferationenzymeactivityprimary
