健骨二仙丸对成骨细胞分化及IGF-ImRNA表达的影响  

‘JianGuErXianWan’ Regulating Human Osteoblast-like Cell Differentiation and IGF- ImRNA Expressing

作　　者：程志安[1] 吴燕峰[1] 曾志勇[1] 谢文峰[1] 沈慧勇[1] 萧劲夫 

机构地区：[1]中山大学附属第二医院骨科,广东广州510120 [2]深圳市中医药研究所

出　　处：《中国中医骨伤科杂志》2004年第2期4-9,共6页Chinese Journal of Traditional Medical Traumatology & Orthopedics

基　　金：国家中医药管理局科研基金资助课题;编号 :97CO42

摘　　要：目的 :在临床研究和动物实验的基础上 ,进一步探讨健骨二仙丸对成骨细胞分化的影响及其作用机制。方法 :分离、培养人的原代成骨细胞 ,采用 PNPP法、RIA法、茜素红染色法以及定量 RT-PCR法分别检测碱性磷酸酶活性、骨钙素含量、矿化结节形成和 型前胶原 m RNA的表达 ,从而了解健骨二仙丸对成骨细胞分化的影响。定量 RT-PCR和 Northern印迹杂交等方法交观察成骨细胞胰岛素样生长因子 ( insulin-like growth factor-I) IGF-Im RNA的表达。结果 :健骨二仙丸中、高剂量能显著增加碱性磷酸酶的活性 ,及矿化结节形成的数量 ,促进成骨细胞骨钙素的释放 ( P<0 .0 1 ,P<0 .0 5 ) ,与雌激素 (倍美力 )组比较差异无显著性。显著增加成骨细胞 型前胶原 m RNA和 IGF-Im RNA的表达。结论 :健骨二仙丸促进成骨细胞 IGF- m RNA的表达 ,从而进一步促进成骨细胞的增殖、分化 ,改善成骨细胞功能 ,调节骨重建偶联。表明健骨二仙丸不仅能增加成骨细胞胶原蛋白地合成 ,而且能促进非胶原蛋白地表达。促进骨的形成是健骨二仙丸作用的主要机制。Objective:Research the effects of ‘JianGuErXianWan’on the differentiation of the human osteoblast-like cells (HOBCs) and the mechanisms of the drug on modulating proliferation,differentiation and preventing apoptosis of the cells.Methods:HOBCs were isolated and cultured according to the references.The differentiation of human osteoblsat-like cells was observed by the activity of Alkaline Phosphatase (ALP) and Oeteocalcin, the number of mineral nodules,and expressing of Procollogen Ⅰ α mRNA.The methods of PNPP, radioimmunoassay,alizarin staining and Quantitative reverse transcript polymerase chain reactions (RT-PCR) were used respectively.RT-PCR and Northern blot hybridization were applied to detect the Insulin-like Growth factor-Ⅰ (IGF-Ⅰ) mRNA expressing.Results:The activity of ALP and osteocalcin of ‘JianGuErXianWan’ groups of the moderate and high dosages are significantly higher than that of control groups.The numbers of mineral nodules,and the expressing of procollagen Ⅰ α mRNA of the two groups are also significantly enhanced than that of control groups.The drug also has a positive effect on the expressing of IGF-ImRNA in HOBCs.Conclusion:‘JianGuErXianWan’ has an effectively action on the differentiationand IGF-mRNA expressing of HOBCs in vitro. These indicate that the drug can not only facilitate the expressing of collagen protein but also non-collagen protein.

关 键 词：成骨细胞 分化 I型前胶原 胰岛素样生长因子-I 健骨二仙丸 

分 类 号：R681[医药卫生—骨科学] Q813[医药卫生—外科学]