黄芪对脂肪源性干细胞成骨作用的初步探讨

Effect of Huangqi on the Differentiation of Adipose-derived Stem Cells into Osteoblasts

目的探索黄芪对脂肪源性干细胞(ASCs)成骨能力的影响,为诱导ASCs向成骨分化提供一个更好的体外环境。方法复苏人ASCs细胞,当细胞生长至融合时传代,取第3代人ASCs用成骨诱导液进行培养,取第4代用于实验。实验共分为三组:空白组(完全培养基干预ASCs)、诱导组(成骨诱导液干预ASCs)和黄芪组(黄芪注射液与成骨诱导液按体积1.5∶8.5配制干预ASCs)。采用MTT检测细胞增殖情况;RT-PCR检测成骨相关指标碱性磷酸酶(ALP)和I型胶原(Collagen I)mRNA的表达。结果 MTT表明在早期(3d)诱导组和黄芪组均促进了细胞增殖,与空白组相比差异有统计学意义(P<0.05);随着干预时间的延长,诱导组和黄芪组细胞增殖速度反低于空白组,空白组增殖情况明显优于诱导组和黄芪组(P<0.05)。RT-PCR表明诱导组和黄芪组均可促进ALP(0.42±0.02)、(0.61±0.01)和Collagen I(0.39±0.09)、(0.56±0.18)的表达,与空白组(0.23±0.06)、(0.11±0.03)相比差异有明显的统计学意义(P<0.05);黄芪组与诱导组相比差异也有明显的统计学意义(P<0.05)。结论黄芪具有明显的促进干细胞向成骨细胞分化的作用,为黄芪参与骨缺损修复提供了一定的实验资料。

Objective To explore the effect of Huangqi on the differentiation of adipose-derived stem cells（ASCs） into osteoblasts which will provide a comfortable environment to induced the differentiation into osteoblasts.Methods ASCs were recovered and passaged.The third passage of ASCs was used for this experiment.The whole experiment was divided into the following groups： the control group,the induction group and the huangqi group.MTT was used to test the proliferation of ASCs and RT-PCR was used to test the mRNA expression of ALP and Collagen I.Results MTT showed that the proliferation of ASCs was significantly promoted by both the induction and the Huangqi groups,compared with the control group（P〈0.05） in the first time point and it was the contrary in the late period（P〈0.05）.RT-PCR showed that the expression of ALP（0.42±0.02,0.61±0.01） and Collagen I（0.39±0.09,0.56±0.18） increased significantly in the induction and the Huangqi groups,compared to the control groups（0.23±0.06,0.11±0.03）（P〈0.05）.And the Huangqi group significantly increased the expression of ALP and Collagen I compared to the induction group（P〈0.05）.Conclusion Huangqi may increase the osteoblastic ability of ASCs,which will provide a basis of theory by using the Chinese Medicine to repair bone defect.