Study on the mechanism of Wumen Gumi Bao decoction containing serum in promoting mesenchymal stem cells osteogenic differentiation based on Wnt/β-catenin signaling pathway
Study on the mechanism of Wumen Gumi Bao decoction containing serum in promoting mesenchymal stem cells osteogenic differentiation based on Wnt/β-catenin signaling pathway
Objective To study mechanism of Wumen Gumi Bao decoction containing serum in promoting mesenchymal stem cells osteogenic differentiation based on Wnt/β-catenin signaling pathway.Methods SD rat bone marrow mesenchymal stem cells were divided into the normal serum group,the classic inducer[mesenchymal stem cell Osteoblast Medium(OBM)]group,Wumen Gu Mi Bao decoction containing serum low dose(10g crude drug/kg)and high dose(20g crude drug/kg)group,the inhibitor[secreted protein Dickkopf-1(DKK-1),0.1ng·mL-1]group,and Wumen Gu Mi Bao decoction containing serum low and high dose+the inhibitor groups.CCK8 method was used to detect the activity of mesenchymal stem cells after 7 days of intervention after osteogenic differentiation.And then the ALP staining intensity and activity of mesenchymal stem cells stained with alkaline phosphatase(ALP)staining method were statistically analyzed,and these mesenchymal stem cells were intervened by the different doses of Wumen Gu Mi Bao decoction containing serum,and Wnt/β-catenin signaling pathway specific blocker DKK-1.The levels of 3αlpha(Wnt3α),β-Catenin,glycogen synthase kinase 3β(GSK 3β),osteoclastogenesis inhibitory factor(OCIF),also known as osteoprotegerin(OPG),and receptor activator of nuclear factor-κB ligand(RANKL)in the Wnt signaling protein family of mesenchymal stem cells were determined by Western Blot.Results Compared with the normal group,intervention with the different doses of Wumen Gu Mi Bao decoction containing serum could effectively promote the proliferation of mesenchymal stem cells(P＜0.05);Wumen Gu Mi Bao decoction containing serum could effectively promote the intensity and activity of ALP staining in mesenchymal stem cells(P＜0.05),as well as regulate the expression levels of osteogenic differentiation-related Wnt3α,β-Catenin,GSK3β,OPG and RANKL proteins(all P＜0.05),but these regulatory effects could be reversed by DKK-lto a certain extent.Conclusion Wumen Gu Mi Bao decoction may induce the osteogenic differentiation of bone marrow mesenchymal stem cells by activating Wnt/β-catenin signaling pathway and affecting OPG/RANKL pathway.
关键词
骨质疏松/吴门骨密葆方/Wnt/β-catenin通路/OPG/RANKL通路/成骨分化
Key words
osteoporosis/Wumen Gu Mi Bao decoction/Wnt/β-catenin pathway/OPG/RANKL pathway/osteogenic differentiation
维普
万方数据