胰高血糖素样肽-1对人脂肪干细胞增殖、成脂分化及脂代谢的影响

Effects of glucagon like peptide-1(GLP-1) on the proliferation, adipogenesis and lipid metabolism of human adipose-derived stem cells

目的研究胰高血糖素样肽-1(GLP-1)对人脂肪干细胞(ASCs)增殖、成脂分化及脂代谢的影响。方法胶原酶分离法原代培养人ASCs,体外扩增及传代建立稳定的培养体系。用含有不同浓度GLP-1(0、0.1、1.0、10.0及100.0 nmol/L)的基础培养基和成脂分化培养基培养细胞,噻唑蓝法(MTT)检测干预24、48及72 h后细胞增殖情况,分化21 d后行油红O染色、异丙醇萃取法检测细胞内脂质含量,同时测定分化过程中上清液中甘油释放量作为脂质分解的指标,测定细胞裂解液中三酰甘油的含量作为脂质合成指标。结果 1高浓度的GLP-1(100.0 nmol/L)对人ASCs的增殖具有抑制作用,且随着时间的延长抑制作用更明显。2油红O染色及异丙醇萃取法显示100.0 nmol/L的GLP-1抑制人ASCs的成脂分化。3在人ASCs成脂分化过程中,高浓度的GLP-1(100.0 nmol/L)可促进甘油的释放,而对细胞内三酰甘油的含量影响不大。结论高浓度GLP-1可抑制人ASCs的增殖及成脂分化,促进脂质分解,对脂质合成无明显作用。

[Objective] To investigate the effects of GLP-1 on the proliferation, adipogenesis and the lipid metabolism in the course of the adipogenesis of human adipose-derived stem cells （ASCs）. [Methods] Human ASCs were separated from subcutaneous adipose tissue by collagenase I and cultured and induced to adipogenesis in the presence of GLP-1 of different concentrations （0, 0.1, 1.0, 10.0, 100.0 nmol/L） Oil red O staining and isopropanol extraction methods were used to assess intracellular lipid accumulation. Supernatant of cell culture medium was collected every 3 day to measure glycerol as lipolysis index, cell lysate was collected to measure triglyceride as lipid synthesis index. [ Results ] Human ASCs were successfully cultured from subcutaneous adipose tissue, and can be induced to osteogenic, adipogenic and chondrogenic differentiation in vitro. MTF results showed that the proliferative activity of human ASCs was attenuated by GLP-1 on higher concentration [A（0.430 ± 0.052） vs （0.290 ±0.060）, P 〈 0.01]. Oil red 0 staining showed that GLP-1 on higher concentration （100.0 nmol/L） inhibited the differentiation of human ASCs[A（0.542 ± 0.023） vs （0.618 ± 0.017）, P 〈 0.05]. During the course of the human ASCs adipogenesis, GLP-1 （100 nmol/L）accelerated the release of glycerol [（398.75±25.19） μmol/L vs （163.44 ± 45.23） μmol/L, P 〈 0.011, but had no effect on the content of intracellular triglyceride accumulation[（10.503±0.259） mmol/L/g vs （11.090 ± 0.281） mmol/L/g, P 〉 0.05]. [Conclusion] GLP-1 can inhibit the proliferation and adipogenesis of human ASCs in vitro, promote lipolysis, but has no effect on lipid synthesis.