CAJ | 학술논문

目的研究淫羊藿苷对体外培养成人骨髓基质干细胞(human bone marrow stromal stem cells,hBMSCs)增殖与成骨性分化的影响.方法取住院患者术后骨髓标本(男,40岁),利用骨髓细胞处理试剂盒分离单核层细胞,培养于含10%FBS的DMEM培养液中,3 d后首次换液,12 d后传代培养.培养基中淫羊藿苷终浓度分别为5×10~(-5)、1×10~(-5)、5×10~(-6)、1×10~(-6)、5×10~(-7)、1×10~(-7) moL/L.增殖分析采用MTT法,于成骨性诱导培养第8d测碱性磷酸酶(alkaline phosphatase,ALP)活性,第14 d进行茜素红染色及钙化结节计数.结果原代培养细胞呈典型成纤维细胞样形态;淫羊藿苷剂量依赖性抑制hBMSCs增殖,但能显著促进其向成骨性分化,表现为提高hBMSCs的ALP活性,增加钙化结节数量.结论终浓度为5×10~(-5) mol/L淫羊藿苷显著促进hBMSCs的成骨性分化,证明淫羊藿苷是中药淫羊藿抗骨质疏松的有效成分.
목적 연구음양곽감대체외배양성인골수기질간세포(human bone marrow stromal stem cells,hBMSCs)증식여성골성분화적영향.방법 취주원환자술후골수표본(남,40세),이용골수세포처리시제합분리단핵층세포,배양우함10%FBS적DMEM배양액중,3 d후수차환액,12 d후전대배양.배양기중음양곽감종농도분별위5×10~(-5)、1×10~(-5)、5×10~(-6)、1×10~(-6)、5×10~(-7)、1×10~(-7) moL/L.증식분석채용MTT법,우성골성유도배양제8d측감성린산매(alkaline phosphatase,ALP)활성,제14 d진행천소홍염색급개화결절계수.결과 원대배양세포정전형성섬유세포양형태;음양곽감제량의뢰성억제hBMSCs증식,단능현저촉진기향성골성분화,표현위제고hBMSCs적ALP활성,증가개화결절수량.결론 종농도위5×10~(-5) mol/L음양곽감현저촉진hBMSCs적성골성분화,증명음양곽감시중약음양곽항골질소송적유효성분.
Objective To investigate the effects of icariin on human bone marrow stromal stem cells in vitro under the conditions of the ability to differentiate into osteoblasts and the case of proliferation.Methods Obtained the inpatient's (male,42 years old) bone marrow sample,used the bone marrow cell treatment kits to separate and collect the stratum of mononuclear cells.The cells cultured in containing 10% fetal bovine serum DMEM.Three days later changed the culture medium at the first time.Twelve days later,proceeded serial subcultivation.The icariin final concentration are 5 ×10~(-5),1×10~(-5),5×10~(-6),1×10~(-6),5×10~(-7),1 ×10~(-7) mol/L.Proliferated analysis adopted MTT method.Under the induce condition,the Alkaline phosphatase activity measured at the eighth day.At the fourteenth day,proceeded alizarin red stain and counted Calcified tubercle.Results Primary culture cells submitted typical fibroblast form.The hBMSCs's proliferation refrained by icariin of dose dependent.But it significantly advanced to osteogenesis.Manifested raised the ALP activity and increased Calcified tubercle amount.Conclusion The icariin with final concentration 5×10~(-5) mol/L can predominantly promote hBMSCs differentiation to osteogenesis.Demonstrated the icariin can prophylaxis antiosteoporotic,which is an active constituent of the traditional Chinese medicine epimedium.