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ID:54601293
大小:1.34 MB
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时间:2020-05-03
《一贯煎诱导骨髓间充质干细胞分化为肝细胞样细胞的实验研究-论文.pdf》由会员上传分享,免费在线阅读,更多相关内容在行业资料-天天文库。
1、中国中西医结合杂志2014年3月第34卷第3期CJITWM,March2014,Vo1.34,No.3一贯煎诱导骨髓问充质干细胞分化为肝细胞样细胞的实验研究平键陈红云。周扬成扬徐列明,摘要目的体外实验观察一贯煎诱导分离培养的大鼠骨髓间充质干细胞(bonemarrowmesen—chymalstemcell,BMSC)定向分化肝细胞的作用。方法采用全骨髓贴壁法分离原代大鼠BMSC,第3代细胞采用流式细胞仪分析表面细胞因子、诱导分化为脂肪细胞鉴定其干细胞特征;采用肝细胞生长因子联合成纤维细胞生长因子和一贯煎含药血清2种诱导方案连续培养BMSC共21天
2、,PCR检测细胞甲胎蛋白(仅一fetoprotein,AFP)、白蛋白(albumin,AIb)、肝细胞核因子4or(hepatocytenuclearfactor4e,HNF4仅)mRNA表达情况,细胞免疫荧光检测细胞AFP和细胞角蛋白18(cytokeratin18,CK18)的表达,染色高碘酸一希夫反应法观察细胞糖原形成,Westernblot法检测细胞CK18、Wnt35、oL.catenin蛋白的表达情况。结果全骨髓贴壁法分离的BMSC高表达白细胞分化抗原(clusterofdifferentiation,CD)90、29、44,低表达
3、CD34和CD11b,成脂诱导2周后油红O染色可见大量脂滴。一贯煎药物血清和细胞因子诱导均可提高BMSC表达AIb、AFP、HNF4o~mRNA表达水平,促进CK18蛋白表达,诱导21天后可检测到AFP和CK18的表达,可见细胞合成糖原颗粒,观察到BMSC内Wnt3or和B—catenin表达下调。结论一贯煎可诱导大鼠BMSC向肝细胞样细胞分化,其机制与下调Wnt/13.catenin信号通路有关。关键词一贯煎;骨髓间充质干细胞;转分化EfectofYiguanDecoctiononDiferentiationofBoneMarrowMesenc
4、hymaIStemCellsintoHepa—tocyte-likeCells:anExperimentaIResearchPINGJian‘’。,CHENHong—yun。,ZHOUYang。,CHENGYang。’,andXULie—ming1InstituteofLiverDiseases,ShuguangHospitalAffiliatedtoShanghaiUniversityofTraditionalChineseMedicine,Shanghai(201203),China;2KeyLaboratory,Min-istrvofEdu
5、cationonLiverandKidneyDiseases。ShanghaiUniversityofTraditiona
6、ChineseMedicine。Shanghai(201203),China;3DepartmentofTraditionalChineseMedicine,ChanghaiHospital,SecondMilitaryMedicalUniversity,Shanghai(200433),China;4DepartmentofLiverCirrhosis,ShuguangHos-pitalAffiliatedtoShangh
7、aiUniversityofTraditionafChineseMedicine。Shanghai(201203),ChinaABSTRACTObiectiveToobservetheeffectofYiguanDecoction(YGD)ondi仟erentiationofbonemarrowmesenchymalstemcells(BMSCs)intohepatocyte—likecellsfnvitro.MethodsRatBM—SCswereisolatedusingwholebonemarrowadherentmethod.Thepro
8、pertiesofBMSCswereidentifiedbyanalyzingtheexpressionofsurfacecytokinesbyflowcytometry.Thethirdpassagecellsweredifferentia—tedintofatcellstoidentifytheirfeatures.BMSCswereincubatedwithhepatocytegrowthfactor(HGF)plusfibroblastgrowthfactor4(FGF4)orYGDcontainingserumYGDfor21days.
9、ThemRNAexpressionofor..fetoprotein(AFP),albumin(AIb),andhepatocytenu
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