体外诱导条件下大鼠骨髓基质细胞骨钙素和Ⅰ型胶原mRNA的表达

目的 研究体外培养大鼠骨髓基质细胞诱导条件下，成骨分化标志骨钙素和Ⅰ型胶原mRNA水平的表达。方法 采用成骨诱导剂（含地塞米松10^-8mol/L,β甘油磷酸钠10mmol/L和抗坏敌国酸AA50μg/ml)对培养状态下的不同代大鼠骨髓基质细胞进行成骨诱导，提取RNA，采用RT-PCR方法，以β-肌动蛋白cDNA为内参照，检测骨钙素和Ⅰ型胶原mRNA的表达。结果 与对照组相比，接受成骨诱导剂作用的大鼠骨髓基质细胞随诱导时间的延长骨钙素和Ⅰ型胶原mRNA出现高表达，且维生素D为诱导骨钙素mRNA表达的必需成分。结论 体外培养的大鼠骨髓基质细胞保持成骨未分化状态，无骨钙素和Ⅰ型胶原mRNA的表达，诱导后可向成骨分化，骨钙素和Ⅰ型胶原mRNA表达升高。

Osteocalcin and type I collagen mRNA expression of rat marrow stromal cells under induction condition]

OBJECTIVE: To investigate osteocalcin and type I collagen mRNA expression of rat marrow stromal cells under induction condition. METHODS: Cultured rat marrow stromal cells were induced by osteogenic compound(composed of dexamethasone 10(-3) mol/L, beta glyceralphosphate 10 mmol/L, ascorbic acid 50 micrograms/ml). Osteocalcin and type I collagen mRNA expression were measured using RT-PCR technique. RESULTS: It was found that rat marrow stromal cells in induced group have an increased time-dependent expression of osteocalcin and type I collagen mRNA, in comparison with control groups. And the necessity of 1,25-(OH)2D3 in induced osteocalcin mRNA expression was confirmed. CONCLUSION: The cultured rat marrow stromal cells kept the undifferentiated state and had osteogenic potential testified by increased osteocalcin and type I collagen mRNA expression when induced.