人参皂苷Rb1对体外培养人脂肪干细胞增殖与成骨分化的影响(英文)

Ginsenoside Rb1 affects the proliferation and osteogenic differentiation of human adipose-derived stem cells in vitro

背景:脂肪干细胞成骨分化受多种因素的影响,中药成骨诱导活性因子在脂肪干细胞研究中有重要意义。目的:观察人参皂苷Rb1在体外培养条件下对人脂肪干细胞增殖和成骨分化的影响。方法:体外分离培养人脂肪干细胞,传至第3代后,按2×103/孔接种至96孔板,分别加入0.5,1.0,2.0,4.0,6.0μmol/L人参皂苷Rb1培养基200μL进行培养;设立对照组,仅加入等量普通DMEM培养基。采用XTT比色法测定大鼠脂肪干细胞的生长增殖曲线。通过碱性磷酸酶试剂盒测定细胞碱性磷酸酶活性,放射免疫法检测骨钙素含量,茜素红染色观察钙化结节形成能力。结果与结论:0.5μmol/L人参皂苷Rb1可明显促进人脂肪干细胞增殖;随着人参皂苷Rb1浓度的增加,促细胞增殖活性降低,6.0μmol/L人参皂苷Rb1表现为明显的抑制细胞增殖作用。人参皂苷Rb1呈剂量依赖性促进人脂肪干细胞碱性磷酸酶活性和骨钙素表达。4.0,6.0μmol/L人参皂苷Rb1诱导钙化结节形成能力优于0.5,1.0和2.0μmol/L人参皂苷Rb1,对照组人脂肪干细胞未见钙化结节形成。提示人参皂苷Rb1在一定浓度范围内对体外培养条件下的人脂肪干细胞具有促生长增殖作用,但在高浓度时,人参皂苷Rb1对人脂肪干细胞的成骨分化具有促进作用,因此可作为一种良好的成骨诱导活性因子。

BACKGROUND： Various factors can affect the osteogenic differentiation of human adipose-derived stem cells, and the osteoinductive factor of traditional Chinese medicine is very important for the research of human adipose-derived stem cells. OBJECTIVE： To investagate the effects of ginsenoside Rb1 on the proliferation and osteogenic differentiation of human adipose-derived stem cells in vitro. METHODS： The human adipose-derived stem cells were isolated and cultured in vitro. After passaqed to the third generation, human adipose-derived stem cells at 2×10 3 /well were incubated in a 96-well plate, and treated with 200 μL of 0.5, 1.0, 2.0, 4.0,6.0 μmol/L ginsenoside Rb1 medium. The human adipose-derived stem cells in the control group were treated with an equal volume of Dulbecco’s modified Eagle medium. Growth curves were examined by 2,3-bis-（2-methoxy-4-nitro-5-sulphenyl）-（2H）-tetrazolium-5-carboxanilide T colorimetric assay. Alkaline phosphatase activity and osteocalcin content were detected by alkaline phosphatase kit and radio-immunity method, respectively. Calcified nodules were observed using alizarin red O staining. RESULTS AND CONCLUSION： The proliferation viability of human adipose-derived stem cells was significantly increased after cultured with 0.5 μmol/L ginsenoside Rb1. With the increasing of the concentration of ginsenoside Rb1, the mitogenic activity of the cells was decreased. The 6.0 μmol/L ginsenoside Rb1 showed a depressant effect on proliferation. Ginsenoside Rb1 could promote alkaline phosphatase activity and osteocalcin expression in human adipose-derived stem cells and showed a dose-dependent manner. Calcified nodule formation induced by 4.6 and 6.0 μmol/L ginsenoside Rb1 were better when compared with 0.5, 1.0 and 2.0 μmol/L ginsenoside Rb1. Ginsenoside Rb1 can promote the proliferation of human adipose-derived stem cells cultured in vitro in a certain concentration, and in the high concentration, the ginsenoside Rb1 can promote the osteogenic differentiation of human adipose-derived stem cells. So ginsenoside Rb1 can be used as an osteoinductive factor.