黄芪甲苷联用维生素 D3对二维回转培养大鼠原代成骨细胞增殖的影响

摘要

探讨了黄芪提取物黄芪甲苷与维生素 D3联用对二维回转培养大鼠原代成骨细胞增殖的影响。离体培养大鼠原代成骨细胞后,用碱性磷酸酶染色、茜素红染色等方法对细胞进行鉴定；通过细胞回转器模拟微重力效应,采用MTT、细胞计数等方法研究不同浓度配比的黄芪甲苷与维生素 D3混合液对大鼠原代成骨细胞增殖的影响。结果表明,单用黄芪甲苷或维生素 D3均对正常培养成骨细胞增殖具有促进作用,最佳促增殖浓度分别为1×10-7 mol·L-1和1×10-8 mol·L-1,当1×10-7 mol·L-1黄芪甲苷分别配以1×10-9 mol·L-1或1×10-8 mol·L-1维生素 D3时,其相对增殖率可达200%和196%；不同浓度配比的黄芪甲苷与维生素 D3混合液对模拟微重力下二维回转培养成骨细胞的增殖有显著促进作用,其最佳促增殖浓度为1×10-7 mol·L-1黄芪甲苷配以1×10-9 mol·L-1维生素 D3。%Effect of astragaloside Ⅳ combined with vitamin D3 on the proliferation of rat primary osteoblasts during 2D-clinorotation culture was investigated.Cultured rat primary osteoblasts were obtained and identified by using alkaline phosphatase staining and alizarin red staining methods.Clinostat were used to simulate micro-gravity,and the effects of astragaloside Ⅳ combined with vitamin D3 at different concentration ratios on prolif-eration of rat primary osteoblasts were studied by MTT assay and cell counting method.Results showed that, during normal culture,the proliferation of rat primary osteoblasts was significantly increased by treating with astragaloside Ⅳ or vitamin D3 ,and the optimal concentration was 1×10 -7 mol·L-1 astragaloside Ⅳ or 1×10 -8 mol· L-1 vitamin D3 ,respectively.Moreover,the proliferative effect was further enhanced when osteoblasts were treated with astragaloside Ⅳ(1×10 -7 mol·L-1 )combined with vitamin D3 (1×10 -9 mol·L-1 or 1×10 -8 mol·L-1 ),and the relative proliferation rate was 200% and 1 96%,respectively.During 2D-clinorotation cul-ture,the proliferation of rat primary osteoblasts were significantly increased by mixed solutions of astragalosideⅣ combined with vitamin D3 at different concentration ratios and the optimal conditions were 1×10 -7 mol·L-1 astragaloside Ⅳ combined with 1×10 -9 mol·L-1 vitamin D3 .