胰岛素样生长因子-1对奶牛成骨细胞增殖及分化的影响

Effect of insulin-like growth factor-1 on the proliferation and differentiation of osteoblastic cells from Holstein cattle

研究了外源性胰岛素样生长因子-1(IGF-1)对体外培养的奶牛成骨细胞增殖、分化及矿化的影响。应用组织块移行法分离奶牛成骨细胞,姬姆萨染色及碱性磷酸酶(ALP)染色进行鉴定。MTT法检测不同质量浓度IGF-1对成骨细胞增殖的影响,比色法检测及放射免疫法(RIA)分别测定细胞基质ALP活性及骨钙素(OC)水平,评价细胞分化功能。结果显示,1～200μg/LIGF-1均能促进奶牛成骨细胞增殖,100μg/L组具有最大刺激效应,且IGF-1干预组细胞光密度值(D值)从第1天到第5天逐渐增加,第7天降低。与对照组相比,10μg/L与100μg/LIGF-1均可显著增强ALP活性及OC水平,增幅达20%～180%(P<0.05或者P<0.01)。结果表明,IGF-1是奶牛成骨细胞增殖和分化代谢促进剂,因此推测IGF-1可以成为动物骨代谢性疾病潜在治疗手段。

This study investigated the effect of exogenous insulin-like growth factor （IGF）-1 on the proliferation and differentiation of osteoblastic cells from Chinese Holstein cattle in vitro.The osteoblastic cells were isolated and cultured,then identified using Giemsa and alkaline phosphatase （ALP） staining methods.The effect of different concentrations of IGF-1 on cell growth was assessed by MTT assay.The ALP activity and osteocalcin （OC） concentration in the osteoblastic cells were measured by a colorimetric assay and a radioimmmunoassay,respectively.Cell proliferation in the cultures was stimulated by IGF-1 at concentrations ranging from 1-200 μg/L,with the a peake at 100 μg/L at day 5,and then decreased at day 7.At 10 μg/L and 100 μg/L,IGF-1,ALP activity and OC level significantly induced by 20% to 180% （P0.05 or P0.01）,compared to the controls.The results suggested that IGF-1 is an anabolic agent for proliferation and differentiation of dairy cow osteoblasts,therefore could act as a potential treatment for the metabolic bone diseases in the animals.