七厘散含药血清对骨髓间充质干细胞增殖、成骨分化的影响

目的 分析七厘散对体外培养的骨髓间充质干细胞增殖，成骨分化的影响。方法 采用5倍等效剂量七厘散灌服大鼠，制得含药血清，以灌服等体积生理盐水制得对照血清；采用贴壁筛选法培养大鼠骨髓间充质干细胞，同时进行细胞表型鉴定，并以含药血清干预第三代rBMSCs。MTT 法检测细胞增殖情况；于干预后的第3、6、9和12d分别测定细胞碱性磷酸酶（alkaline phosphatase，ALP）活性、干预后6、11、16d分别测定骨钙素（BGP）、Ⅰ型胶原、钙盐沉积量及钙化结节数，并比较两组间差异。结果 七厘散含药血清显著促进外源性骨髓间充质干细胞增殖及促进骨髓间充质干细胞成骨性分化，表现在该组的碱性磷酸酶活性、骨钙素、Ⅰ型胶原、钙盐沉积量和钙化结节数高于对照组（P<0.01）。结论 七厘散含药血清具有显著促进rBMSCs增殖和成骨性分化活性的作用。

Effect of Qilisan containing serum on the proliferation and osteogenic differentiation of rat mesenchymal stem cells

Objective To investigate the effect of Qilisan containing serum on the proliferation and osteogenic differentiation of rat bone mesenchymal stem cells. Methods The drug containing serum was obtained by giving rats a gavage of 5 fold equivalent doses of Qilisan. The control serum was obtained by giving a gavage of equal volume normal saline. Rat bone mesenchymal stem cells (rBMSCs) were selected with adherence method. Cell phenotype was identified at the same time. The drug containing serum was additioned into the culture medium to intervene in the third generation of BMSCs. The proliferation of rBMSCs was analyzed using MTT method. The activity of alkaline phosphatase (ALP) was tested on day 3, 6, 9, and 12 after the intervention. The concentrations of bone gamma-carboxyglutamic-acid-containing proteins (BGP), Col-I, calcium deposition, and the number of mineralized bone modules were tested on day 6, 11, and 16 after the intervention. The results between the two groups were compared. Results Qilisan containing serum significantly promoted the proliferation of exogenous BMSCs and the osteogenic differentiation of BMSCs. The activity of ALP, the concentrations of BGP, Col-I, calcium deposition, and the number of mineralized bone nodules were significantly improved in the treatment group compared to those in control group (P<0.01). Conclusion Qilisan containing serum can significantly promote the proliferation and osteogenic differentiation of rBMSCs.