人参皂甙Rg1对体外培养猪骨髓基质细胞增殖的影响

背景既往的研究表明,人参总甙对体外培养的骨髓造血干细胞及祖细胞有促进增殖作用,对骨髓造血因子具有促分泌作用,但对骨髓基质细胞的增殖作用研究较少.目的观察人参皂甙Rg1对体外培养猪骨髓基质细胞增殖的影响.设计对照观察.单位南京医科大学第一附属医院心内科、皮肤科,南京医科大学药理教研室.材料实验于2002-01/2003-02在南京医科大学心血管药理实验室完成.选择实验用猪及人参皂甙Rg1干粉.方法将体外培养的骨髓基质细胞分为对照组及实验组,实验组加入不同浓度的人参皂甙Rg1(10-7,5×10-7,10-6,5×10-6mol/L)诱导培养,对照组加入等量培养基.分别应用成纤维细胞体外集落形成法、[3H]TdR掺入法和四甲基偶氮唑盐法检测骨髓基质细胞的增殖情况.主要观察指标①人参皂甙Rg1对体外培养骨髓基质细胞的成纤维样集落形成单位生长的影响.②人参皂甙Rg1对骨髓基质细胞[3H]TdR掺入的影响.③人参皂甙Rg1对四甲基偶氮唑盐法测定骨髓基质细胞增殖的影响.结果①人参皂甙Rg1对体外培养骨髓基质细胞的成纤维样集落形成单位生长的影响实验组成纤维细胞体外集落形成法集落数目均高于对照组(P＜0.05～P＜0.01),尤以(5～50)×10-7 mol/L组为明显,在浓度为10-6 mol/L集落数即达高峰.②人参皂甙Rg1对骨髓基质细胞[3H]TTdR掺入的影响在人参皂甙Rg1浓度为10-6 mol/L时,其骨髓基质细胞[3H]TdR掺入率明显高于对照组(P＜0.05～P＜0.01),随着剂量的增加其促进增殖作用增强.③人参皂甙Rg1对四甲基偶氮唑盐法测定骨髓基质细胞增殖的影响实验组的光密度值明显高于对照组(P＜0.05～P＜0.01),并呈剂量依赖性.结论人参皂甙Rg1对体外培养猪骨髓基质细胞具有增殖作用,且具有剂量依赖性.

Effect of ginsenoside Rg1 on multiplication of pig''''s bone marrow stromal cells in vitro

BACKGROUND:It is shown in past researches that total saponin of ginseng has the effect of promoting multiplication of bone marrow hematopoietic stem cells and progenitor cells in vitro, and promoting secretion of bone marrow hemotopoietic factors, however there has been little researches on the effect of multiplication of bone marrow stromal cells.OBJECTIVE: To investigate the effect of ginsenoside Rg1 on pig's bone marrow stromal cells multiplication in vitro.DESIGN: A controlled observation.SETTING:Department of Cardiology and Department of Dermotology,First Affiliated Hospital, Nanjing Medical University; Department of Pharmacology, Nanjing Medical University.MATERIALS: The experiment was conducted at the Cardiovascular Pharmaceutical Laboratory of Nanjing Medical University from January 2002 to February 2003. The experimental pigs ahd the powder of ginsenoside Rg1were chosen METHODS :The bone marrow stromal cells cultured in vitro were divided as control and experimental groups. For experimental group insenoside Rg1 in different concentrations (10-7,5×10-7,10-6,5×104 mol/L)were added for induction culture, and for control group the media of the same volume was added. The multiplication conditions of bone marrow stromal cells were respectively detected by fibroblast colony forming in vitro, tritiated thymidine {[3H]TdR} incorporation and MTT.MAIN OUTCOME MEASURES: ① The effect of ginsenoside Rgl on the fibroblast colony forming units of bone marrow stromal cells in vitro. ②The effect of ginsenoside Rg1 on [3H]TdR incorporation of bone marrow stromal cells. ③ The effect of ginsenoside Rg1 on bone marrow stromal cells multiplication detected by MTT.RESULTS:① The effect of ginsenoside Rg1 on the fibroblast colony forming units of bone marrow stromal cells in vitro: The numbers of units in experimental group were all higher than that in control group (P ＜ 0.05-P ＜ 0.01),especially obvious in (5-50)×10-7 mol/L experimental group, reaching peak in the concentration being 10-6 mol/L. ② The effect of ginseno side Rg1 on [3H]TdR incorporation of bone marrow stromalcells: As the concentration of ginsenoside Rg1 was 10-6 mol/L, the [3H]TdR incorporation efficiency of bone marrow stromal cells was obviously higher that that in control group (P ＜ 0.05-P ＜ 0.01), and the effect was enhanced as the dosage increased. ③ The effect of ginsenoside Rgl on bone marrow stromal cells multiplication detected by MTT: The value of absorbance in experimental group was obviously higher than that in control group (P ＜ 0.05-P ＜ 0.01), and it was dose-dependent. CONCLUSION: Ginsenoside Rg1 had an effect of multiplication on pig's bone marrow stromal cells in vitro, and it was dose-dependent.