目的观察不同浓度巴戟天含药血清对体外培养成骨-破骨细胞共育体系中原癌基因(C-fOS)、核心结合因子(CbfA)1 MrnA表达的影响。方法提取24 H内新生Sd乳鼠颅盖骨分离培养成骨细胞,采用5周龄Sd大鼠双侧股骨、胫骨的骨髓基质细胞,加入集落细胞刺激因子(M-CSf)和细胞核因子κb受体活化因子配体(rAnkl)诱导培养破骨细胞。采用碱性磷酸酶(AlP)染色鉴定成骨细胞,抗酒石酸酸性磷酸酶(TrAP)染色、骨吸收陷窝甲苯胺蓝染色、电镜扫描等鉴定破骨细胞,体外建立成骨-破骨细胞共育体系,设置低、中、高三种浓度巴戟天含药血清组和不含药血清组,干预3 d后提取各组总rnA,应用逆转录-聚合酶链反应(rT-PCr)方法测定各组C-fOS、CbfA1 MrnA表达量。结果不同浓度巴戟天含药大鼠血清对成骨-破骨细胞共育体系C-fOS有抑制作用,对CbfA1 MrnA的表达有促进作用。高浓度含药血清组两者的表达差异显著(P<0.05,P<0.000 1)。结论巴戟天含药血清可抑制成骨-破骨细胞共育体系C-fOS的表达,促进CbfA1 MrnA的表达,从而达到降低破骨细胞分化成熟及骨吸收活性,促进骨形成。Objective To observe the effects of the serum of Morinda officinalis( RMO) on the expression of C-FOS and core binding factor Alpha1( Cbfa1) in osteoblast and osteoclast co-cultured system.Methods Osteoblasts were separated from the cranium of 24 hours newborn SD rat.Bone marrow cells were harvested from bilateral femora and tibiae of five weeks old SD rat,and M-CSF and RANKL were used to induce osteoclast formation.Osteoblast cells were confirmed by alkaline phosphatase( ALP) stain,osteoclast cells were confirmed by tartrate resistant acidphos phatase( TRAP) stain,Toluidine blue stain and bone resorption assay.Osteoblast and osteoclast co-cultured system was established in vitro.Low,middle,high concentrations of serum RMO and control groups were set.Total RNA was extracted after intervention 3 days,C-FOS and Cbfa1 mRNA expression were measured by real-time PCR.Results Different concentrations serum of RMO had inhibitory effect on the expression of C-FOS and enhance the mRNA expression of Cbfa1 mRNA,and the function on both indicated a statistically significant difference at high concentration( P<0.05,P<0.000 1).Conclusions The serum of RMO could down-regulate the expression of C-FOS and up-regulate Cbfa1 mRNA in osteoblast and osteoclast co-cultured system,consequently reduce osteoclast differentiation and activity of bone resorption,enhance bone formation.国家自然科学基金资助项目(No.81272168); 福建省医学创新课题资助项目(No.2012-CXB-32
