当归多糖减轻5-氟尿嘧啶对骨髓基质细胞成骨分化的抑制

Angelica sinensis Polysaccharides Reduce 5-Fluorouracil-Induced Inhibition on Osteogenic Differentiation of Bone Marrow Stromal Cells

该实验探讨当归多糖(ASP)对改善5-氟尿嘧啶(5-FU)所致人骨髓基质细胞成骨与成脂分化失衡的作用。人骨髓基质细胞株HS-5体外培养分为:对照组、ASP组、5-FU组、5-FU+ASP组和5-FU+LiCl组。CCK-8检测细胞抑制率,流式细胞术检测细胞凋亡;成骨与成脂诱导分化实验检测细胞成骨与成脂分化能力,Western blot检测Runx2、PPARγ和β-catenin蛋白表达,RT-PCR检测Runx2、OCN、BMP-2、Osterix、PPARγ和β-catenin mRNA表达。结果表明,与对照组相比5-FU作用HS-5细胞后细胞增殖抑制、凋亡率增加,成骨分化能力减弱、成脂分化能力增强,分化相关信号β-catenin蛋白和mRNA表达降低;相比5-FU组,ASP预处理可减少细胞凋亡;恢复细胞成骨分化能力,成骨相关因子Runx2、OCN、BMP-2和Osterix表达升高;降低细胞成脂分化能力,成脂相关因子PPARγ表达减少;β-catenin信号分子表达增加。结果提示,当归多糖可维持5-FU作用后骨髓基质细胞朝成骨方向分化的能力,其机制可能与激活Wnt/β-catenin信号通路有关。

The aim of the study was to investigate the effect of ASP (Angelica sinensis polysaccharides) on ameliorating the imbalance of osteogenic and adipogenic differentiation of human bone marrow stromal cells induced by 5-FU (5-fluorouracil).Human bone marrow stromal cell line HS-5 cells were cultured in vitro and divided into:Control group,ASP group,5-FU group,5-FU+ASP group and 5-FU+LiCl group.Cell inhibition rate of HS-5 cells was detected by CCK-8 assay,and the cellular apoptosis was analyzed by flow cytometry.The ability of BMSCs for osteogenesis and adipogenic differentiation was detected by osteogenic and adipogenic differentiation assays.The expressions of Runx2,PPARγ and β-catenin protiens were measured by Western blot.The mRNA expressions of Runx2,OCN,BMP-2,Osterix,PPARγ and β-catenin were detected by RT-PCR.The results indicated that compared with the control group,the proliferative capacity of HS-5 cells was inhibited and the apoptosis rate was increased in the 5-FU group.The osteogenesis potential was decreased whereas adipogenesis potential was enhanced.The mRNA and protein levels of differentiation-related signaling molecule β-catenin were decreased.However,compared with the 5-FU group,the apoptosis rate of HS-5 cells was decreased after ASP pretreatment.The osteogenesis ability of HS-5 was restored while the adipogenesis potential was reduced.The expression of osteogenic-related factors including Runx2,OCN,BMP-2 and Osterix was increased whereas the expression of the lipid-related factor PPARγ was decreased.The expression of β-catenin was increased in 5-FU+ASP group.The results suggested that Angelica sinensis polysaccharides protected bone marrow stromal cells against 5-FU via maintaining osteogenesis potential,and the underlying mechanism might be related to activating Wnt/β-catenin signaling pathway.