摘要
目的研究PPAR-gamma信号通路对血管瘤来源的间充质干细胞(Hemangioma derived mesenchymal stem cells,Hem-MSCs)成脂分化的影响。方法本研究以不同的条件培养基培养Hem-MSCs,实验分5组:普通培养基组(A组)、普通培养基+罗格列酮组(B组)、成脂诱导分化培养基组(C组)、成脂诱导分化培养基+罗格列酮组(D组)及成脂诱导分化培养基+罗格列酮+GW9662拮抗剂组(E组)。光镜下观察各组Hem-MSCs的成脂分化过程,以油红O染色观察被诱导细胞中脂滴的面积和数量;采用Western-blot分析各组Hem-MSCs培养2周后的perilipin A表达量,以量化各组细胞脂肪分化程度;以实时定量PCR分析各组成脂分化相关基因(如CEBPA、LPL、PLIN1、PPARG等)在HemMSCs成脂诱导分化过程中不同时间点的表达情况。结果油红O染色显示,成脂诱导分化培养基+罗格列酮组可促进Hem-MSCs的成脂分化过程;Western-blot证实罗格列酮能明显促进成脂诱导分化时perilipin A蛋白的表达;实时定量PCR进一步证实,在诱导分化的第1周及第2周时,罗格列酮对Hem-MSCs的成脂分化相关基因(CEBPA、LPL、PLIN1、PPARG等)的表达均有促进作用;而PPAR-gamma信号通路的拮抗剂GW9662能够阻断上述作用。结论激活PPARgamma信号通路可促进Hem-MSCs在体外的成脂分化能力。
Objective To investigate the effect of PPAR-gamma signaling pathway in the adipogenic differentiation of hemangioma derived mesenchymal stem cells(Hem-MSCs)in vitro.Methods Hem-MSCs were cultured in 5 groups of different media,including group A(ordinary culture medium),group B(ordinary culture medium+rosiglitazone),group C(adipogenic medium),group D(adipogenic medium+rosiglitazone),group E(adipogenic medium+rosiglitazone+GW9662).The adipogenic differentiation of hem-MSCs was observed by light microscope,and the range and number of lipid droplets were observed by oil red O staining.The expression of perilipin A in each group was detected by Western-blot after two weeks of culture to quantify the degree of adipogenic differentiation.The expressions of adipogenesis-related genes(CEBPA,LPL,PLIN1,PPARG)of each group at different time points were detected by real-time quantitative PCR.Results Oil red O staining showed that rosiglitazone could promote the adipogenic differentiation of hem-MSCs in adipogenic medium.Western-blot confirmed that rosiglitazone could significantly promote the expression of perilipin A protein in the induced differentiation group.Real-time quantitative PCR further confirmed that rosiglitazone could promote the expression of adipogenesis-related genes(CEBPA,LPL,PLIN1,PPARG)in hem-MSCs at the first and second weeks.GW9662,an antagonist of PPAR-gamma signaling pathway,could block the above effects.Conclusion Activated PPAR-gamma signaling pathway promotes the adipogenic differentiation of Hem-MSCs in vitro.
作者
徐媛
郭遥
袁斯明
王慜
陈海妮
沈卫民
XU Yuan;GUO Yao;YUAN Siming;WANG Min;CHEN Haini;SHEN Weimin(Department of Plastic Surgery,Jinling Hospital,School of Medicine,Nanjing University,Nanjing 210002,China;Department of Plastic Surgery,Nanjing Children's Hospital,Nanjing 210008,China)
出处
《组织工程与重建外科杂志》
2019年第1期1-4,9,共5页
Journal of Tissue Engineering and Reconstructive Surgery
基金
国家自然科学基金面上项目(81272989)
