肾气丸对SD大鼠骨髓间充质干细胞增殖的影响

The Influence of Shenqi Pills on Proliferation of Bone Mesenchymal Stem Cells of SD Rats

目的:观察不同浓度肾气丸水提液对体外培养的SD大鼠骨髓间充质干细胞(BMSCs)增殖的影响。方法:应用全骨髓贴壁法体外分离培养大鼠BMSCs,传代扩增,倒置显微镜进行细胞形态学观察,免疫组化法检测细胞表面抗原鉴定其表面标志。将BMSCs分为对照组及不同浓度肾气丸水提液干预组,采用MTT比色法观察其增殖情况。结果:采用全骨髓贴壁法获取的BMSCs形态呈长梭形,呈集落样生长,免疫细胞化学鉴定,CD44、CD29阳性,CD34阴性。MTT检测结果显示,与空白对照组相比,肾气丸水提液干预24h后,4、2、1、0.5mg/ml浓度肾气丸干预组吸光度值均增高(P<0.05),其中2mg/mL肾气丸干预组增高明显;0.25mg/mL浓度肾气丸水提液干预组吸光度与空白组相比无差异(P>0.05)。结论:本实验采用全骨髓贴壁法培养的骨髓基质细胞符合BMSCs的生物特性,可以用于BMSCs相关研究。在一定浓度范围内,肾气丸水提液可以提高BMSCs的体外增殖能力。

Objective： To observe the influences of Shenqi pill water extracts on the proliferation of bone mesenchymal stem cells（BMSCs） of SD rats in vitro.Methods： The bone mesenchymal stem cells（BMSCs） of SD rats were isolated with adherent separation method and primarily cultured in vitro,morphologically observed under inverted microscope,and identified with immunohistochemical method.The proliferation of BMSCs were affected by different concentrations of Shenqi pill water extracts which were detected with MTT assay.Results： BMSCs presented a long spindle shape and a colony growth pattern.CD44,CD29 on BMSCs were positive and CD34 was negatively detected by immunohistochemical method.MTT assay showed absorbance of BMSCs treated with Shenqi pills for 24 hours at the dose of 4,2,1,0.5 mg/ml,equivalent to 2~0.5mg/ml crude drug,were significantly higher than that in the control group（P0.05）,especially the 2mg/ml group.Conclusion： BMSCs isolated with adherent separation method in this experiment presents the biological characteristics of BMSCs,which can be used in BMSCs correlated research.Within certain range of concentrations,Shenqi pill water extracts can improve the proliferation activity of BMSCs in vitro.