补肾强督方对骨髓间充质干细胞成骨过程Wnt通路中因子表达的影响

Effect of Bushenqiangdu Formula on expressions of GSK3β,wnt5a and SOST in Wnt pathway during osteogenesis of human bone marrow mesenchymal stem cells

目的:研究补肾强督方含药血清对骨髓间充质干细胞(BMSCs)成骨过程Wnt/β-catenin通路中因子表达的影响。方法:BMSCs细胞分为空白对照组、西药对照组、补肾强督方低中高剂量组,分别进行成骨诱导分化,茜素红染色法检测各组成骨程度,实时荧光定量PCR检测GSK3β、wnt5a和SOST基因mRNA表达,Western blot检测Wnt/β-catenin通路关键因子GSK3β、wnt5a和SOST蛋白表达。结果:空白组BMSCs成骨分化培养后可见明显矿化结节形成,补肾强督方干预组未见明显矿化结节。与空白对照组比较,补肾强督方含药血清组BMSCs中SOST、GSK3β蛋白表达显著升高,Wnt5a蛋白的表达显著降低(P<0.05)。与空白对照组比较,补肾强督方含药血清组BMSCs中SOST、GSK3βmRNA的表达显著升高,Wnt5a mRNA的表达显著降低(P<0.05)。结论:补肾强督方含药血清具有抑制强直性脊柱炎骨化的作用,其机制可能与抑制BMSCs成骨过程中的Wnt/β-catenin信号通路相关。

Objective:To study the effect of Bushenqiangdufang serum on the expressions of factors in Wnt/β-catenin pathway during osteogenesis of human bone marrow mesenchymal stem cells(BMSCs).Methods:BMSCs were divided into control group,medicine control group and Bushenqiangdufang low,medium and high dosage groups for osteogenic differentiation.Alizarin red staining was used to detect the degree of bone composition.GSK3β,SOST and wnt5 a gene expressions were detected by real-time PCR,and GSK3βprotein expression was detected by Western blot.Results:The osteogenesis levels of BMSCs in the serum of Bushenqiangdufang and the western medicine control group were lower than that in the control group.Bushenqiangdufang drug-containing serum can promote the expression of SOST and GSK3β,and inhibit the expression of wnt5 a in BMSCs.Conclusion:Bushenqiangdufang drug-containing serum has the effect of inhibiting AS ossification.The mechanism may be related to the inhibition of Wnt/β-catenin signaling pathway in BMSCs osteogenesis.Whether it acts through this pathway,further experimental evidence is needed.