【摘要】 目的观察鲑鱼降钙素(calcitonin,CT)对离体培养数成骨细胞(osteoblast,OB)的作用,初步探讨CT对OB增殖以及胰岛素样生长因子I(Insulin-like Growth Factor-I,IGF-I)表达的影响,为促进骨修复和再生的临床治疗提供一定的理论依据。方法取新生24hSD乳鼠颅盖骨,采用酶消化法培养OB,经酶解消化后得到原代OB,进行体外培养并鉴定。将体外培养的OB用不同浓度(1×10-12～1×10-8mol/L)的CT处理,采用MTT法(噻唑蓝染色法)观察OB的增殖;Gomori钙-钻法检测碱性磷酸酶(ALP)的表达;RT-PCR方法观察胰岛素样生长因子-I(IGF-I)mRNA的表达。结果经细胞鉴定后证实其系OB;MTT法检测表明,CT浓度为1×10-12mol/L时就可以刺激OB增殖,且随CT浓度升高其增殖能力增加,呈剂量依赖关系;Gomori钙-钻法检测结果表明随着CT浓度的升高OB分化能力增强;经RT-PCR法检测表明,IGF-I mRNA可在乳鼠OB内稳定表达,半定量分析表明,随着CT浓度的升高,IGF-ImRNA的表达量逐渐增加。结论适当浓度的CT可以促进体外培养OB的增殖;部分机制可能与CT促进IGF-ImRNA表达的增加有关。更多还原

【Abstract】 Objective Through observe the role of salmon calcitonin(CT) on osteoblasts in vitro culture rat,we are trying to have a preliminary study of the influence of salmon calcitonin on the proliferation of osteoblasts and the expression of IGF-I,this way could provide a theoretical basis of promoting bone repair and regeneration on clinical treatment.Methods Take the calvarias of 24h SD neonatal rats,with enzyme digestion of osteoblast cells,primary osteoblasts are obtained by enzymatic digestion,cultured and identified osteoblasts,with different concentrations of calcitonin on the osteoblasts cultured in vitro.observed the proliferation of OB by MTT method(MTT staining);Detection of Alkaline phosphatase(ALP) expression by Gomori Calcium-Drilling Method;Observed the Insulin-like growth factor-I(IGF-I)mRNA expression by RT-PCR method.Results Cells were identified and confirmed by the osteoblasts.MTT assay showed that OB proliferation can stimulate at the concentration of 1×10-12mol/L,and with the increased proliferative capacity of CT concentrations increase in a dose-dependent;Gomori calcium-drill assay results show that with the calcitonin concentration increased,the ability of the differentiation of OB is enhanced;Detected by RT-PCR showed that IGF-I mRNA in the neonatal rat stable expression in osteoblasts,Semi-quantitative analysis showed that with increasing concentration of calcitonin,IGF-ImRNA expression gradually increased.Conclusion Appropriate concentration of CT can promote osteoblast cell proliferation;part of the mechanism may be related to CT coule promote the expression of IGF-ImRNA increased.更多还原