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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)×10-7mol/L组为明显,在浓度为10-6mol/L集落数即达高峰。②人参皂甙Rg1对骨髓基质细胞[3H]TdR掺入的影响:在人参皂甙Rg1浓度为10-6mol/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。
BACKGROUND: Previous studies have shown that ginsenosides can promote the proliferation of bone marrow hematopoietic stem and progenitor cells cultured in vitro and promote the secretion of bone marrow hematopoietic factors, but there are few studies on the proliferation of bone marrow stromal cells. Objective: To observe the effect of ginsenoside Rg1 on proliferation of porcine bone marrow stromal cells cultured in vitro. Design: Control observation. Department of Cardiology and Dermatology, the First Affiliated Hospital of Nanjing Medical University, Department of Pharmacology, Nanjing Medical University. MATERIALS: The experiment was performed at the Laboratory of Cardiovascular Pharmacology, Nanjing Medical University from January 2002 to February 2003. The experimental pigs and ginsenoside Rg1 dry powder were selected. Methods: In vitro cultured bone marrow stromal cells were divided into control group and experimental group. The experimental group was induced by different concentrations of ginsenoside Rg1 (10-7, 5×10-7, 10-6, 5×10-6 mol/L). Culture, the control group was added equal amount of medium. The proliferation of bone marrow stromal cells was detected by fibroblasts in vitro colony formation assay, [3H]TdR incorporation and MTT assay. MAIN OUTCOME MEASURES: The effect of ginsenoside Rg1 on the growth of fibroblast-like colony forming units in bone marrow stromal cells cultured in vitro. 2 Effects of ginsenoside Rg1 on [3H]TdR incorporation in bone marrow stromal cells. 3 The effect of ginsenoside Rg1 on the proliferation of bone marrow stromal cells measured by MTT method. RESULTS: 1The effect of ginsenoside Rg1 on the growth of fibroblast-like colony forming units in cultured bone marrow stromal cells in vitro: The number of colonies formed by the in vitro fibroblasts colony formation assay was higher than that of the control group (P<0.05 to P<0.01). (5 ~ 50) × 10-7mol / L group was obvious, in the concentration of 10-6mol / L colonies that reached the peak. 2The effect of ginsenoside Rg1 on incorporation of [3H]TdR in bone marrow stromal cells: When the concentration of ginsenoside Rg1 was 10-6 mol/L, the incorporation rate of [3H]TdR in bone marrow stromal cells was significantly higher than that in the control group (P<0.05). ~ P <0.01), with the increase of the dose it promotes enhanced proliferation. 3 The effect of ginsenoside Rg1 on the proliferation of bone marrow stromal cells measured by MTT: The optical density of the experimental group was significantly higher than that of the control group (P<0.05-P<0.01), and was dose-dependent. CONCLUSION: Ginsenoside Rg1 has a proliferative effect in vitro on porcine bone marrow stromal cells in a dose-dependent manner.