淫羊藿总黄酮对兔脂肪干细胞向尿路上皮细胞增殖分化的影响  

Effects of total flavone of epimedium on proliferation and differentiation of rabbit adipose stem cells into urinary epithelial cells

作　　者：胡少华 赵振理 万智盛 卜威振 陈松强 陆毅群[2] HU Shaohua;ZHAO Zhenli;WAN Zhisheng;BU Weizhen;CHEN Songqiang;LU Yiqun(Hainan Women and Children's Medical Center,Haikou 570206,China;Children's Hospital of Fudan University,Shanghai 201102,China)

机构地区：[1]海南省妇女儿童医学中心,海南海口570206 [2]复旦大学附属儿科医院,上海201102

出　　处：《临床医药实践》2023年第11期828-833,共6页Proceeding of Clinical Medicine

基　　金：海南省妇女儿童医学中心院内科研项目(项目编号:2020YJ05);海南省临床医学中心建设项目(项目编号:琼卫医函[2021]75号);海南省卫生健康行业科研项目(项目编号:19A200048)。

摘　　要：目的:探讨淫羊藿总黄酮对兔脂肪干细胞诱导培养向尿路上皮细胞增殖分化的影响。方法:采用蛋白质印迹法(Western blot)对脂肪干细胞进行鉴定,用MTT法检测增殖活力,然后将以未添加淫羊藿总黄酮的培养基作为对照组,将分别加入0.1μg/mL,1μg/mL,10μg/mL,100μg/mL淫羊藿总黄酮的培养基作为实验组,采用间接共培养对脂肪干细胞进行诱导培养,采用Western blot及实时定量聚合酶链式反应(RT-PCR)鉴定尿路上皮细胞,并对比转化效率。结果:脂肪干细胞呈长梭形,表型鉴定结果显示CD44和Vimentin蛋白阳性表达,且MTT显示细胞活力良好。对兔脂肪干细胞诱导培养成功获得尿路上皮细胞,呈多角形,表型鉴定显示:Up-Ⅰa,Up-Ⅲ,AE1/AE3和CK7蛋白阳性表达,Up-Ⅲ的mRNA阳性表达,且在淫羊藿总黄酮浓度为100μg/mL时表达水平最显著。结论:淫羊藿总黄酮对兔脂肪干细胞向尿路上皮细胞的转化具有促进作用,在一定范围内呈现浓度依赖性,为提高转化效率提供了新的思路。Objective:To investigate the effect of total flavone of epimedium on the proliferation and differentiation of rabbit adipose stem cells cultured into urinary epithelial cells.Methods:The adipose stem cells were identified by Western blot,and the proliferation viability was detected by MTT assay.Then,the adipose stem cells were induced by indirect co-cultured using the medium without the addition of total flavone of epimedium as the control group,and the medium with 0.1μg/mL,1μg/mL,10μg/mL,100μg/mL of total flavone of epimedium as the experimental group,respectively.Western blot and reverse transcription-polymerase chain reaction(RT-PCR)were used to identify the uroepithelial cells and compare the transformation efficiency.Results:The adipose stem cells were long shuttle-shaped,phenotypic identification showed positive expression of CD44 and vimentin protein,and MTT showed good cell viability.The rabbit adipose stem cells were successfully cultured to obtain urinary epithelial cells in polygonal shape,and the phenotypic identification showed positive expression of Up-Ⅰa,Up-Ⅲ,AE1/AE3 and CK7 proteins,and positive expression of mRNA of Up-ⅢmRNA,the expression level was most significant when the concentration of total flavonoids in epimedium was 100μg/mL.Conclusion:Total flavone of epimedium can promote the transformation of adipose tissue derived stromal cells(ADSCs)into urinary epithelial cells in a concentration-dependent manner within a certain range and it provides a new idea for improving conversion efficiency.

关 键 词：脂肪干细胞 尿路上皮细胞 淫羊藿总黄酮 诱导培养 

分 类 号：R97[医药卫生—药品]