二苯乙烯苷对骨髓间充质干细胞增殖和分化的影响及其机制研究

目的：探究二苯乙烯苷（TSG）对体外骨髓间充质干细胞（BMSCs）增殖和分化的影响，并初步探讨其可能的作用机制。方法：利用全骨髓培养法从1月龄SD大鼠中提取BMSCs进行体外培养，然后给予药物干预。采用MTT法检测药物对BMSCs体外增殖的影响；PNPP法检测碱性磷酸酶（ALP）活性；ELISA法检测细胞培养上清液中骨钙素（OCN）含量；实时荧光定量PCR（qRT-PCR）观察Col1a1、Runx2和β-catenin的mRNA表达情况。结果：TSG在1×10^-6，1×10^-5，1×10^-4 mol·L^-1浓度下对BMSCs的增殖有明显的促进作用；经1×10^-6，1×10^-5，1×10^-4 mol·L^-1 TSG处理3 d后，BMSCs中ALP的表达增加；经1×10^-6，1×10^-5，1×10^-4 mol·L^-1 TSG处理7 d后，OCN表达也明显升高；此外，与对照组相比，经Wnt3a和TSG处理后，BMSCs中Col1a1、Runx2和β-catenin的mRNA表达上调。结论：TSG能在一定程度上促进BMSCs的增殖和成骨分化能力，其机制可能与提高Col1a1、Runx2和β-catenin相关成骨分化基因的表达有关。

Effects and mechanism of tetrahydroxystilbene glucoside on the proliferation and differentiation of bone marrow mesenchymal stem cells

OBJECTIVE To investigate the effects of tetrahydroxystilbene glucoside (TSG) on the proliferation and differentiation of bone marrow mesenchymal stem cells (BMSCs) in vitro, and to explore its possible mechanism.METHODS The whole bone marrow culture technique was used to isolate the BMSCs from one-month SD rats, and then drug intervention was given. MTT method was used to measure the effect of TSG on the proliferation of BMSCs in vitro; alkaline phosphatase (ALP) activity was detected by PNPP and the content of osteocalcin (OCN) in the cell supernatant was measured by ELISA. Meanwhile, quantitative real-time PCR(qRT-PCR)was used to detect the mRNA expression of Col1a1, Runx2 and β-catenin.RESULTS TSG significantly promoted the proliferation of BMSCs at the concentration of 1×10^-6,1×10^-5,1×10^-4 mol·L^-1. After 3 days of treatment of TSG at the concentration of 1×10^-6,1×10^-5,1×10^-4 mol·L^-1, the expression of ALP in BMSCs increased. Besides, the expression of OCN were also significantly increased after 7-day treatment. After the treatment of Wnt3a and TSG, the mRNA expression of Col1a1, Runx2 and β-catenin was higher than that in the control group.CONCLUSION TSG can promote the proliferation and osteogenic differentiation of BMSCs, which may be related to the up-regulation of bone formation-related genes including Col1a1, Runx2 and β-catenin.