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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: Osteogenic differentiation of adipose-derived stem cells is influenced by many factors. Osteoinductive factors of traditional Chinese medicine are of great importance in the study of adipose-derived stem cells. Objective: To observe the effects of ginsenoside Rb1 on the proliferation and osteogenic differentiation of human adipose-derived stem cells in vitro. METHODS: Human adipose-derived stem cells were isolated and cultured in vitro. After passage 3, they were inoculated into 96-well plates at 2 × 103 / well and cultured with 200μl of 0.5,1.0,2.0,4.0,6.0μmol / L ginsenoside Rb1 medium ; Set up a control group, adding only the same amount of ordinary DMEM medium. The growth and proliferation curve of rat adipose derived stem cells was determined by XTT colorimetry. Alkaline phosphatase kit was used to measure the activity of alkaline phosphatase. Radioimmunoassay was used to detect the content of osteocalcin. Alizarin red staining was used to observe the formation of calcified nodules. RESULTS AND CONCLUSION: 0.5μmol / L ginsenoside Rb1 can significantly promote the proliferation of human adipose-derived stem cells. With the increase of ginsenoside Rb1 concentration, the cell proliferative activity decreased. 6.0μmol / L ginsenoside Rb1 showed obvious inhibitory effect on cell proliferation. Ginsenoside Rb1 promotes alkaline phosphatase activity and osteocalcin expression in human adipose-derived stem cells in a dose-dependent manner. 4.0, 6.0μmol / L ginsenoside Rb1 induced calcification nodules better than 0.5,1.0 and 2.0μmol / L ginsenoside Rb1, the control group of human adipose stem cells no calcified nodules. It is suggested that ginsenoside Rb1 can promote proliferation and proliferation of human ADSCs in vitro under certain concentration range. However, Ginsenoside Rb1 can promote osteoblastic differentiation of ADSCs at high concentration, Good osteoinductive activity factor.