黄芪多糖对犬骨髓基质干细胞增殖及超微结构的影响

目的探讨黄芪多糖（APS）对诱导培养的犬骨髓基质干细胞（BMSCs）增殖、分化成骨作用及超微结构的影响。方法用瑞氏- 姬姆萨染色、改良Gomori钙- 钴法、茜素红染色法检测BMSCs分化成骨的能力。用四唑盐（MTT）比色和酶动力学方法测定不同浓度和时间的APS对诱导培养的BMSCs增殖影响以及超微结构的改变。结果诱导条件下BMSCs具有成骨细胞活性，瑞氏- 姬姆萨染色呈深蓝色，改良Gomori钙- 钴法将细胞及矿化结节染成黑色，茜素红染色法见细胞及矿化结节成红色。第1、3天时，低浓度（0.005 mg/mL）APS可促进诱导培养的BMSCs增殖；而第5天时，高浓度（50 mg/mL）APS则明显抑制诱导培养的BMSCs增殖。第5天，透射电镜下，0.005 mg/mL APS组细胞线粒体、粗面内质网增多，细胞外基质分泌增加；50 mg/mL APS组细胞发生退变，粗面内质网和线粒体明显减少，且二者明显肿胀、变性、膜结构破坏。结论短期低浓度APS能促进诱导培养的BMSCs的代谢和蛋白质的合成，有利于细胞的增殖和向成骨分化。

Effect of astragalus polysaccharides on the proliferation and ultrastructure of dog bone marrow stem cells induced into osteoblasts in vitro

Objective To observe the growth and osteogenic property of cultured dog bone marrow stem cells(BMSCs) by investigating the effects of astragalus polysaccharides(APS) on the proliferation and ultrastructure of BMSCs into osteoblasts in vitro. Methods BMSCs osteogenic property was detected by improved Wright-Giemsa, Gomori and alizarin dyeing method. The proliferation and differentiation of the induced BMSCs with APS in different concentration and time were detected by MTT assay and the morphologic change of the induced BMSCs was observed by transmission electron microscope(TEM). Results BMSCs osteogenic property was detected with Wright-Giemsa deep-bluing, Gomori method blacking and with more mineral nodules alizarin dyeing method carmining. APS with concentration of 0.005 mg/mL can promote the proliferation of the induced BMSCs in short-term culture(1th, 3th day) and 50 mg/mL can decrease the effect through long-term culture(5th day). Observed by TEM(5th day), the number of mitochondria, rough endoplasmic reticulum increased and the extracellular matrix was excreted more in the induced BMSCs by APS with concentration of 0.005 mg/mL. However, not only the number of mitochondria, rough endoplasmic reticulum reduced but also the structure was swollen, degenerative, membrance damaged in the induced BMSCs by APS with concentration of 50 mg/mL. Conclusion APS with lower concentration in short-term culture may promote BMSCs proliferation and differentiation.