Wnt/β-catenin通路在新北美圣草苷促进BMSCs成骨分化中的介导作用

Mediation of Wnt/β-catenin Pathway in the Promotion of Osteogenic Differentiation of BMSCs by Neoeriocitrin

目的:探究新北美圣草苷激活Wnt/β-catenin通路促进骨髓间充质干细胞(Bone Marrow Mesenchymal Stem Cells,BMSCs)成骨分化的具体机制,为其应用于临床治疗提供理论基础。方法:选取SPF级3月龄大鼠30只,随机分为对照组、低剂量组、高剂量组,每组10只,分别给予生理盐水、0.81g/kg及4.05g/kg骨碎补水煎液灌胃并制备含药血清。培养并鉴定BMSCs。以不同浓度含药血清干预BMSCs并比较各组的细胞增殖能力,矿化能力,β-catenin、Lrp-5、Osterix、RUNX-2、GSK-3βmRNA表达水平及β-catenin蛋白、Lrp-5蛋白水平。结果:与对照组相比,给药两组各时间点的细胞增殖能力更强,且高剂量组优于低剂量组;与对照组相比,给药两组的矿化结节数量明显升高,且高剂量组矿化结节数量多于低剂量组;给药两组的GSK-3βmRNA表达明显低于对照组,且高剂量组低于低剂量组,β-catenin、Lrp-5、Osterix、RUNX-2 mRNA表达明显高于对照组,且高剂量组高于低剂量组;同时给药两组的β-catenin蛋白、Lrp-5蛋白水平明显高于对照组,且高剂量组高于低剂量组,差异均具有统计学意义(P<0.05)。结论:新北美圣草苷可能通过激活Wnt/β-catenin通路,上调β-catenin、Lrp-5、Osterix、RUNX-2 mRNA表达并下调GSK-3βmRNA表达,同时上调β-catenin蛋白、Lrp-5蛋白水平,进而促进BMSCs增殖和成骨分化。

Objective:To explore the specific mechanism of neoeriocitrin activating the Wnt/β-catenin pathway to promote the osteogenic differentiation of BMSCs,so as to provide a theoretical basis for its application in clinical treatment.Methods:30 SPF-grade three-month-old rats were randomly divided into the control group,low-dose group and high-dose group,with 10 rats in each group.The rats were intragastrically administered with normal saline,0.81g/kg and 4.05g/kg Rhizoma Drynariae decoction respectively to prepare the medicated serum.Culture and identify BMSCs.The BMSCs were intervened with different concentrations of serum containing drugs,and the cell proliferative capacity,mineralization capacity,mRNA expression levels ofβ-catenin,Lrp-5,Osterix,RUNX-2,and GSK-3βas well as the protein levels ofβ-catenin and Lrp-5 of each group were compared.Results:Compared with the control group,the cell proliferation abilities of the two groups for medicine at each time point were stronger.Compared with the control group,the number of mineralized nodules in the administration groups were significantly increased,and the number of mineralized nodules in the high-dose group was more than that in the low-dose group.The expression of GSK-3βmRNA in the two groups after administration of drug was significantly lower than that in the control group,and the expression in the high-dose group was lower than that in the low-dose group.The expression levels ofβ-catenin,Lrp-5,Osterix and RUNX-2 mRNA in the two groups after administration were significantly higher than those in the control group,and higher in the high-dose group than that in the low-dose group.At the same time,the levels ofβ-catenin and Lrp-5 protein in the two groups after medication were significantly higher than those in the control group,and the levels in the high dose group were higher than those in the low dose group,with statistical significance(P<0.05).Conclusion:Neoeriocitrin may activate the Wnt/β-catenin pathway and up-regulate the expressions ofβ-catenin,Lrp-5,Osterix and RUNX-2 mRNA and down-regulate the expression of GSK-3βmRNA,and up-regulate the levels ofβ-catenin and Lrp-5 protein,which will promote the proliferation and osteogenic differentiation of BMSCs.