CAJ | 학술논문

目的:研究淫羊藿苷对成骨细胞增殖、分化以及对核心结合因子α1(Cbfαl)、骨形成蛋白-2(BMP2)、骨形成蛋白-4(BMP4)mRNA表达的影响.方法:以酶序列消化法从新生大鼠颅骨分离、培养成骨细胞,采用碱性磷酸酶染色和钙结节茜素红染色法鉴定细胞.取第3-5代细胞,用CCK-8法检测经0 mol/L、10~(-8)mol/L、10~(-7)mol/L、10~(-6)mol/L、10~(-5)mol/L、10~(-4)mol/L的淫羊藿苷处理24、48、72 h后成骨细胞的增殖情况.分别用流式细胞技术和对硝基苯酚法检测上述各浓度的淫羊藿苷处理48 h后成骨细胞的增殖指数和碱性磷酸酶(ALP)活性.用real-timePCR法检测10～(-6)mol/L淫羊藿苷处理48 h后成骨细胞Cbfα1、BMP2和BMP4 mRNA表达的改变.结果:10~(-8)～10~(-4)moL/L的淫羊藿苷对成骨细胞均无增殖促进作用,但可促进成骨细胞的ALP活性;10~(-6)mol/L淫羊藿苷可以上调成骨细胞Cbfα1、BMP2和BMP4 mRNA的表达.结论:淫羊藿苷可能是通过上调Cbfα1、BMP2和BMP4 mRNA的表达而促进成骨细胞的分化.
목적:연구음양곽감대성골세포증식、분화이급대핵심결합인자α1(Cbfαl)、골형성단백-2(BMP2)、골형성단백-4(BMP4)mRNA표체적영향.방법:이매서렬소화법종신생대서로골분리、배양성골세포,채용감성린산매염색화개결절천소홍염색법감정세포.취제3-5대세포,용CCK-8법검측경0 mol/L、10~(-8)mol/L、10~(-7)mol/L、10~(-6)mol/L、10~(-5)mol/L、10~(-4)mol/L적음양곽감처리24、48、72 h후성골세포적증식정황.분별용류식세포기술화대초기분분법검측상술각농도적음양곽감처리48 h후성골세포적증식지수화감성린산매(ALP)활성.용real-timePCR법검측10～(-6)mol/L음양곽감처리48 h후성골세포Cbfα1、BMP2화BMP4 mRNA표체적개변.결과:10~(-8)～10~(-4)moL/L적음양곽감대성골세포균무증식촉진작용,단가촉진성골세포적ALP활성;10~(-6)mol/L음양곽감가이상조성골세포Cbfα1、BMP2화BMP4 mRNA적표체.결론:음양곽감가능시통과상조Cbfα1、BMP2화BMP4 mRNA적표체이촉진성골세포적분화.
Objective: To investigate the effect of icariin on the proliferation, differentiation, and the mRNA expressions of Cbfαl, BMP2, BMP4 of rat osteoblasts. Methods: Primary rat osteoblastic cells were obtained by sequentia collagenase/trypsin enzyme digestion from calvarial bones of new born ( within 24 h) SD rats and were identified by Alkaline phosphatase and alizarin red staining. The passage 3-5cells were treated with icariin at the concentration of 0 mol/L, 10~(-8)mol/L, 10~(-7)mol/L, 10~(-6)mol/L,10~(-5)mol/L, 10~(-4)mol/L for 24 h, 48 h, 72 h, and the proliferation of the cells was measured by CCK-8assay. The proliferation index was detected by Flow Cytometry and the activity of alkaline phosphatase was determined by p-Nitrophenyl phosphate (pNPP) method after being treated with icariin at the concentration mentioned above for 48 h. The total cellular RNA was extracted 48 h after being treated with icariin at the concentration of 10~(-6)mol/L, and the expressions of Cbfα1, BMP2, BMP4 mRNA were examined by real-time PCR. Results: Icariin showed no effect on the proliferation of osteoblasts, but improved ALP activity. The Cbfα1, BMP2, BMP4 mRNA were significantly upregulated after icariin treatment. Conclusion: Icariin could promote the differentiation ability of rat osteoblasts through upregulating the Cbfα1, BMP2, BMP4 mRNA expressions.