组织工程与重建外科杂志

组织工程与重建外科杂志 ›› 2019, Vol. 15 ›› Issue (1): 1-9.doi: 10.3969/j.issn.1673-0364.2019.01.001

• 论著 •    下一篇

PPAR-gamma信号通路对Hem-MSCs体外成脂分化的影响

徐媛，郭遥，袁斯明，王慜，陈海妮，沈卫民   

  1. 中国人民解放军南京总医院整形外科；南京市儿童医院整形外科
  • 收稿日期:2018-11-10 修回日期:2018-12-23 出版日期:2019-02-20 发布日期:2019-02-20
  • 作者简介:共同第一作者:徐媛;共同第一作者:郭遥;袁斯明(E-mail:yuansm@163.com)。

The Effect of PPAR-Gamma Signaling Pathway in the Adipogenic Differentiation of Hemangioma Derived Mesenchymal Stem Cells in Vitro

XU Yuan,GUO Yao,YUAN Siming,WANG Min,CHEN Haini,SHEN Weimin   

  1. 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)
  • Received:2018-11-10 Revised:2018-12-23 Online:2019-02-20 Published:2019-02-20
  • Contact: 国家自然科学基金面上项目(81272989)

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。

关键词: 婴幼儿血管瘤, 血管瘤来源的间充质干细胞, 成脂分化, PPAR-gamma信号通路, 罗格列酮

Abstract: 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.

Key words: Infantile hemangioma, Hemangioma derived mesenchymal stem cells, Adipogenic differentiation, PPAR-gamma signaling pathway, Rosiglitazone

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