(论文)神经生长因子诱导肝星状细胞凋亡及其相关机制的研究_[完稿]

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1、•论著°神经生长因子诱导肝星状细胞凋亡及其相关机制的研究舒建昌邓延梅朱海燕呂霞何雅军陈莲香叶国荣【摘要】目的观察神经生长因子(nervegrowthfactor,NGF)对大鼠肝星状细胞(hepaticstellatecelLHSC)凋亡的影响，并探讨其可能的作用机制。方法体外培养大鼠肝星状细胞株HSC-T6,将HSC-T6与100ng/mlNGFW育24h后,流式细胞术检测细胞凋亡;细胞免疫化学法检测HSC中凋亡相关基因P^'BclJ'CaspaseB蛋白表达的悄况;免疫荧光法检测NGF、神经生长因子低亲和力受体p75NTR表达的悄况。结果NGF作用HSC后凋亡率较

2、对照组明显增高[(22.36±9.51)%V8(5.88±1.36)%,P<0.05]o凋亡相关蛋白尸)、3吟・3的阳性细胞百分率与对照组比较明显增高[(78.41±4.00)%、(39.26±1.57)%vs(34.96±3.84)%.(9.27±1.01)%,P<0.05],而Bcl・2表达实验组阳性细胞百分率较对照组降低[(18.12±1.38)%vs(91.53±2.98)%,P<0.05]oNGF作用HSC后NGF表达增多(6.53±1.40vs1.77±0.17,P<0.05),而p75NTR表达无明显变化(3.52±0.36v84.24±0.38,P>0

3、.05)o结论NCF可能通过使凋亡相关基因卩'、3呼・3表达上调、Bd・2表达下调•而诱导HSC凋亡。NGF作用HSC后可作为始动因子和效应因子增加NGF的表达•而对P75NTR表达无影响。[关■词]神经生长因子/药理学;肝细胞/药物作用/病理学;细胞凋亡/药物作用StudyonthemechanismofapoptosisofhepaticstellatecellsinducedbynervegrowthfactorSHUJian-chang,DENGYan-mei,ZHUHai-yan,LVXiatHEYa~jun,CHENLian~xiangtYEGuo^ron

4、g.DepartmentofGastroenterologyfGuangzhouRedCrossHospital9MedicalCollege9JinanUniversity^Guangzhou5102201China[Abstract)ObjectiveToinvestigatetheeffectofNGFonapoptosisofHSCinvitroandexplorethepossiblemechanism.MethodsHSCwasincubatedwithdifferentconcentrationsofNGF.HSCapoptosiswasidentifi

5、edbyFCM・Theexpressionsofapoptosis-regulatingproteinsCaspase-39P53andBcl*2ofHSCafterapoptosisinducedbyNGFwereexaminedbyimmunohistochemicalstaining.ExpressionsofNGFandp75NTRweredetectedbyimmunofluorescence・ResultsApoptosisindexofHSCwashigherthanthatofcontrolgroup[(22.36±9.51)%vs(5.88±1.36

6、)%]aftertreatedwithNGF(100ng/ml)(P<0.05).Afterincubatingwith100ng/mlNGFfor24h9thepositiveexpressionratesofP53andCaspase-3ofHSCincreasedsignificantlythanthoseofcontrolgroup[(7&41±4.00)%vs(34.96±3.84)%f(39.26±1.57)%vs(9.27±1.01)%tP<0.05].ThepositiveexpressionrateofBel-2proteinofHSCsignifi

7、cantlydecreasedcomparedwiththatofcontrolgroup(18.12±1.38)%vs(91.53±2.98)%(P<0.05).WhenHSCwasstimulatedwith100ng/mlNGFfor24httheaverageopticaldensityofNGFincreasedsignificantlythancontrolgroup(6.53±L40vs1.77±0.17)(P<0.05)fwhiletheexpressionofp75NTRwasnotsignificantlychanged(3.