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时间:2020-05-04
《HepG2细胞表达蛋白HCVC体外活化肝星状细胞的研究-论文.pdf》由会员上传分享,免费在线阅读,更多相关内容在应用文档-天天文库。
1、甲国痢原生彳刁_子芾心2013年9月_弟卺帚IU期JournalofPathogenBiologyOctober2013,Vo1.8,No.10·869··论著·HepG2细胞表达蛋白HCVC体外活化肝星状细胞的研究*孙英梅,李文杰,陶剑平~(1.青岛市妇女儿童医院新筛实验室,山东青岛266034;2.中山大学中山医学院,广东广州510080)【摘要】目的舰察HCV核心蛋白对肝星状细胞活化及合成细胞外基质的影响,为进一步研究HCV核心蛋白的致肝纤维化机制提供实验依据。方法采用双抗体夹心ElISA检测HepG2一HCVC细胞株培养液中TGFq31的
2、表达。用HepG2一HCV—C和HepG2两株细胞培养5d的上清液培养人肝星状细胞系Ix2细胞,同时用DMEM培养IX一2细胞作对照,分别制备细胞爬片,采用免疫细胞化学染色(ICC)法检测IX~2细胞中人平滑肌肌动蛋白(a—SMA)、IV型胶原(ColⅣ)、结缔组织生长冈子(CTGF)和纤维连接蛋白(FN)的表达;采用双抗体夹心EIISA检测I一2细胞1~6d培养上清液中人ColⅣ、Ⅲ型前胶原肽(PⅢNP)、透明质酸(HA)和人层粘连蛋白(LN)的表达。所有数据采用SPSS11.0统计软件进行统计分析。结果双抗体夹心EIISA检测HepG2一HC
3、V—c细胞1~6d培养液中的TGFH1量为(11O.O0±l11.45)pg/ml~(935.O0±2I.36)pg/ml,HepG2细胞为0~(124.16±11.81)pg/ml,差异有统计学意义(F一21984.81,P4、epG2)组和Ix一2(DMEM)组(均P5、ntheactivationofhepaticstellatecellsinvitrobyHepG2cellsexpressingHCVcoreproteinSUNYing—mei,LIWen—jie,TAOJian—ping(1.QingdⅡ0WomennndChildre”sHo£n£,Qingdn0266034,China;2.ZhongshanSchoolofMedicine,SunYatsenUniversity,Guangzhou510080,China)[Abstract]ObjectivesToobservetheimpactof6、HCVcoreproteinontheactivationofhepaticstellatece11s(HSCs)andsynthesisofextracellularmatrix(ECM)inordertoprovideanexperimentalbasisforfurtherstudyofthemechanismoffi—brogenesisinducedbyHCVcoreprotein.MethodsTheexpressionofTGFB1byculturedHepG2一HCVCandHepG2cellswasdeterminedusing7、doubleantibodysandwichElISA.Inaddition,thesupernatantofthetwoce11lineswasusedtocultivateLX~2cells.LX2cellsculturedwithDMEMservedasacontro1.Immunocytochemistry(ICC)wasusedtode—tecttheexpressionofa~SMA,ColⅣ,CTGF,andFNinIX一2cellsculturedfromthetwocellslinesandcontrolIX一2cellscul8、turedwithDMEM.DoubleantibodysandwichELISAwasusedtodetecttheexpressio
4、epG2)组和Ix一2(DMEM)组(均P5、ntheactivationofhepaticstellatecellsinvitrobyHepG2cellsexpressingHCVcoreproteinSUNYing—mei,LIWen—jie,TAOJian—ping(1.QingdⅡ0WomennndChildre”sHo£n£,Qingdn0266034,China;2.ZhongshanSchoolofMedicine,SunYatsenUniversity,Guangzhou510080,China)[Abstract]ObjectivesToobservetheimpactof6、HCVcoreproteinontheactivationofhepaticstellatece11s(HSCs)andsynthesisofextracellularmatrix(ECM)inordertoprovideanexperimentalbasisforfurtherstudyofthemechanismoffi—brogenesisinducedbyHCVcoreprotein.MethodsTheexpressionofTGFB1byculturedHepG2一HCVCandHepG2cellswasdeterminedusing7、doubleantibodysandwichElISA.Inaddition,thesupernatantofthetwoce11lineswasusedtocultivateLX~2cells.LX2cellsculturedwithDMEMservedasacontro1.Immunocytochemistry(ICC)wasusedtode—tecttheexpressionofa~SMA,ColⅣ,CTGF,andFNinIX一2cellsculturedfromthetwocellslinesandcontrolIX一2cellscul8、turedwithDMEM.DoubleantibodysandwichELISAwasusedtodetecttheexpressio
5、ntheactivationofhepaticstellatecellsinvitrobyHepG2cellsexpressingHCVcoreproteinSUNYing—mei,LIWen—jie,TAOJian—ping(1.QingdⅡ0WomennndChildre”sHo£n£,Qingdn0266034,China;2.ZhongshanSchoolofMedicine,SunYatsenUniversity,Guangzhou510080,China)[Abstract]ObjectivesToobservetheimpactof
6、HCVcoreproteinontheactivationofhepaticstellatece11s(HSCs)andsynthesisofextracellularmatrix(ECM)inordertoprovideanexperimentalbasisforfurtherstudyofthemechanismoffi—brogenesisinducedbyHCVcoreprotein.MethodsTheexpressionofTGFB1byculturedHepG2一HCVCandHepG2cellswasdeterminedusing
7、doubleantibodysandwichElISA.Inaddition,thesupernatantofthetwoce11lineswasusedtocultivateLX~2cells.LX2cellsculturedwithDMEMservedasacontro1.Immunocytochemistry(ICC)wasusedtode—tecttheexpressionofa~SMA,ColⅣ,CTGF,andFNinIX一2cellsculturedfromthetwocellslinesandcontrolIX一2cellscul
8、turedwithDMEM.DoubleantibodysandwichELISAwasusedtodetecttheexpressio
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