Ming Leiguo, Chen Keming, Ge Baofeng, Ma Huiping, Zhou Jian
Institute of Orthopaedics, Lanzhou General Hospital, Lanzhou 730050, China.
Zhongguo Zhong Yao Za Zhi. 2011 Aug;36(16):2240-5.
To investigated the effect of icariin and genistein on proliferation and mineralization of cultured rat osteoblasts (rat calvarial osteoblasts, ROB). And to contrast the pharmacological activity of icariin and genistein.
Bone cells were obtained by enzyme digestion from the segregated neonatal SD rat skull, and were cultured in MEM containing 10% FBS which was changed after three days later. Serial subcultivation was proceeded when cells covered with 90% culture dish. The final action concentration of icariin and genistein were both 1 x 10(-5) mol x L(-1). Proliferation was analyzed by MTT on 96-well plates, while differentiation was analyzed on 24-well plates. Under the induced condition, the alkaline phosphatase activity, calcium salt sediment yield and osteocalcin were measured at the 3, 6, 9, 12 d. At 12th day, ALP staining, alizarin red staining and calcified nodule count were preceded. Total RNA was isolated at 0, 6, 12, 24, 48, 72 h. The gene expression of bFGF, IGF-1, Osterix and Runx-2 was analyzed by Real-time RT-PCR.
With the concentration of 1 x 10(-5) mol x L(-1), icariin and genistein have no significant effect on the ROB' s proliferation. The osteogenesis, ALP activity, calcium salt sediment yield and osteocalcin, calcified tubercle amount were significantly increased. And they enhanced the mRNA level of bFGF, IGF-1, Osterix and Runx-2. On the level of osteoblasts, the activity of icariin is stronger than that of genistein.
When the final concentration of icariin and genistein is 1 x 10(-5) mol x L(-1), they can significantly promoted ROB maturation. And on the level of osteoblasts, the activity of icariin is stronger than that of genistein.
研究淫羊藿苷和染料木黄酮对培养的大鼠成骨细胞（大鼠颅骨成骨细胞，ROB）增殖和矿化的影响，并比较淫羊藿苷和染料木黄酮的药理活性。
采用酶消化法从新生SD大鼠分离的颅骨中获取骨细胞，接种于含10%胎牛血清的MEM培养基中培养，3天后换液。细胞长满培养皿90%时进行传代培养。淫羊藿苷和染料木黄酮的终浓度均为1×10⁻⁵mol/L。用MTT法在96孔板上分析细胞增殖情况，在24孔板上分析细胞分化情况。诱导培养条件下，分别于3、6、9、12天检测碱性磷酸酶活性、钙盐沉积量和骨钙素含量。第12天进行碱性磷酸酶染色、茜素红染色和钙化结节计数。分别于0、6、12、24、48、72小时提取总RNA，采用实时荧光定量逆转录聚合酶链反应（Real-time RT-PCR）分析碱性成纤维细胞生长因子（bFGF）、胰岛素样生长因子-1（IGF-1）、osterix和Runx-2基因的表达。
终浓度为1×10⁻⁵mol/L时，淫羊藿苷和染料木黄酮对ROB的增殖无显著影响，但可显著增加其成骨能力、碱性磷酸酶活性、钙盐沉积量、骨钙素含量和钙化结节数量，并上调bFGF、IGF-1、osterix和Runx-2的mRNA水平。在成骨细胞水平上，淫羊藿苷的活性强于染料木黄酮。
淫羊藿苷和染料木黄酮终浓度为1×10⁻⁵mol/L时，均可显著促进ROB成熟，且在成骨细胞水平上，淫羊藿苷的活性强于染料木黄酮。
