龙鳖胶囊对骨髓间充质干细胞增殖活性的影响

Effect of Longbie Capsule on proliferation of bone marrow mesenchymal stem cells

背景:临床应用补肾活血中药龙鳖胶囊治疗骨关节炎取得满意的疗效,但其作用机制尚不明确。目的:观察龙鳖胶囊对大鼠骨髓间充质干细胞增殖活性的影响。方法:采用全骨髓贴壁培养方法分离培养SD大鼠骨髓间充质干细胞,倒置显微镜进行形态学观察,流式细胞术检测细胞表面标志表达。用含龙鳖胶囊的完全培养基培养第4代骨髓间充质干细胞,药物质量浓度分为5,1 g/L和250,50,10 mg/L,作用24 h后用MTT法测定细胞活力。结果与结论:原代细胞呈现不规则形状贴壁生长,第2代细胞呈典型的纤维状,第3代细胞呈长纤维状、漩涡式生长。第4代骨髓间充质干细胞的表面分子CD29、CD90呈强阳性表达,CD44呈阳性表达,CD45、CD34呈阴性表达。5 g/L和1 g/L质量浓度的龙鳖胶囊溶液可促进骨髓间充质干细胞增殖。结果表明龙鳖胶囊可能是通过促进骨髓间充质干细胞的增殖和分化,进而发挥治疗骨关节炎的作用。

BACKGROUND：Longbie Capsule has satisfactory outcomes in the treatment of osteoarthritis, but its mechanism is stil unclear. OBJECTIVE：To study the effect of Longbie Capsule on proliferation of bone marrow mesenchymal stem cel s. METHODS：The bone marrow mesenchymal stem cel s from SD rats were separated and expanded by whole adherence culture, then subcultured and confirmed by morphological observation and flow cytometry. Passage 4 cel s were cultured in complete media containing 5 g/L, 1 g/L, 250 mg/L, 50 mg/L, 10 mg/L Longbie Capsule, respectively, for 24 hours. Then, MTT assay was used to detect cel viability. RESULTS AND CONCLUSION：The primary cel s were adherent cel s characterized by irregular shape, passage 2 cel s were typical y fibrous-shaped, passage 3 cel s grew in long fibrous and swirl-type shape. Passage 4 cel s were strongly positive for CD29 and CD90, positive for CD44, and negative for CD34 and CD45. 5 g/L and 1 g/L Longbie Capsule promoted the proliferation of bone marrow mesenchymal stem cel s. These findings indicate that Longbie Capsule may promote the proliferation and differentiation of bone marrow mesenchymal stem cel s,thereby playing a therapeutic effect on osteoarthritis.