摘要
目的 观察金雀异黄酮对缺氧体外培养大鼠骨髓间充质干细胞（BMSCs）增殖与成骨分化的影响。方法 采用贴壁筛选法体外培养BMSCs并用三气培养箱建立缺氧模型。设常氧对照组、缺氧对照组和缺氧加药组，其中缺氧加药组细分为1×10-8、1×10-7和1×10-6 mol·L-1组金雀异黄酮组，缺氧处理36 h后分析各组的细胞增殖和成骨性指标包括碱性磷酸酶（ALP）活性、Ⅰ型胶原表达、钙盐沉积量和钙化结节面积等，并用Real Time RT-PCR法检测Runx2和骨形成蛋白（BMP）-2的基因表达情况。结果 与缺氧对照组比较，金雀异黄酮抑制BMSCs增殖，显著提高ALP活性、Ⅰ型胶原表达量、钙盐沉积量和钙化结节面积，并提高基因Runx2和BMP-2的表达水平，且呈现出剂量依赖性特点。结论 金雀异黄酮具有抑制缺氧BMSCs增殖并促进其成骨分化的作用。
Abstract
OBJECTIVE To investigate the effects of genistein on proliferation and osteogenic differentiati of rat bone mesenchymal stem cells（BMSCs） treated with hypoxia. METHODS Rat BMSCs were cultured by adherence screening method and hypoxic environment was made by triple-gases incubator. At 36 hours after hypoxia，proliferation and osteogenic differentiation markers including alkaline phosphatase （ALP） activity，collagenⅠ，calcium salt sediment yield and mineralized bone nodules were compared among 1×10-8， 1×10-7， 1×10-6 mol·L-1group，Hypoxia Control and Normal Control. Total RNA was isolated and the gene expression of Runx2 and bone morphogenetic proteins-2 （BMP-2） was investigated by Real Time RT-PCR. RESULTS Genistein decreased proliferation and improved ALP activity，expression of collagenⅠ，calcium salt sediment yield and area of mineralized nodules significantly. Besides，the relative level of Runx2 and BMP-2 mRNA expression were enhanced. CONCLUSION Genistein can inhibited proliferation and enhance osteogenic differentiation of BMSCs treat with hypoxia significantly.
关键词
金雀异黄酮 /
骨髓间充质干细胞 /
缺氧 /
成骨性分化 /
增殖 /
Ⅰ型胶原 /
碱性磷酸酶 /
核心结合因子a1
{{custom_keyword}} /
Key words
genistein /
bone mesenchymal stem cells /
hypoxia /
osteogenic differentiation /
proliferation /
collagenⅠ /
alkaline phosphatase /
core binding factor a1 /
morphogenetic proteins-2
{{custom_keyword}} /
韩桂秋 葛宝丰 陈克明 马慧萍.
金雀异黄酮对体外培养缺氧大鼠骨髓间充质干细胞增殖与成骨分化的影响[J]. 中国药学杂志, 2011, 46(16): 1248-1253
HN Gui-qiu;GE o-feng;CHEN Ke-ming;M Hui-ping.
Effect of Genistein on Proliferation and Differentiation of Rat Bone Mesenchymal Stem Cells Treated with Hypoxia in Vitro[J]. Chinese Pharmaceutical Journal, 2011, 46(16): 1248-1253
{{custom_sec.title}}
{{custom_sec.title}}
{{custom_sec.content}}
参考文献
［1］ KEN E，MURAGLIA A，CORSI A，et al. Autologous bone marrow stromal cells loaded onto porous hydroxyapatite ceramic accelerate bone repair in critical-size defects of sheep long bones［J］. J Biomed Mater Res，2000，49（3）：328-337.
［2］ PETITE H，VIATEAU V，BEOSAID W，et al. Tissue-engineered bone regeneration［J］. Nat Biotechnol，2000，18（9）：959-963.
［3］ ZHAO R，ISE H，HONGO M，et al. Human amniotic mesenchymal cells have some characteristics of cardiomyocytes［J］. Transplantation，2005，79（5）：528-535.
［4］ GRUNDES C，REIKERAS O. Blood flow and mechanical properties of healing bone： femoral osteotomies studied in rats ［J］. Acta Orthop Scand，1992，63（5）： 487-453.
［5］ FOTI P，ERBA D，RISO P，et al.Comparison between daidzein and genistein antioxidant activity in primary and cancer lymphocytes［J］.Arch Biochem Biophys，2005，433（2）：421-427.
［6］ MING L G，GE B F，CHEN K M，et al. Effects of osthol on bone marrow stromal stem cells differentiate and proliferate in vitro［J］. Chinese Pharmacological Bulletin，2010，26（8）：1098-1103.
［7］ YAMAMOTO N，FURUYA K，HANADA K，et al. Progressive development of the osteoblast phenotype during differentiation of osteoprogenitor cells derived from fetal rat calvaria： model for in vitro bone formation ［J］. Biol Pharm Bull，2002，25 （4）： 509-515.
［8］ THOMAS D，SCHMITTGEN L，KENNETH J. Analyzing real-time PCR data by the comparative CT method ［J］. Nature Protocol，2008，13（6）：1101-1108.
［9］ GORDILLO G M，SEN C K. Revisiting the essential role of oxygen in wound healing ［J］. Am J Surg，2003，186（3）：259-263.
［10］ ARNETT T R，GIBBONS D C，UTTING J C，et al. Hypoxia is a major stimulator of osteoclast formation and bone resorption［J］. Cell Physiol，2003，196（1）： 2-8.
［11］ MICHAEL M，LYNNE J，DAVID H. Nontraumatic osteoneerosis of the femoral head： ten years later［J］. J Bone Joint Surg Am，2006，88（5）：1117-1132.
［12］ MALLADI P，XU Y，CHIOU M，et al.Effect of reduced oxygen tension on chondrogenesis and osteogenesis in adipose derived mesenchymal cells ［J］. Am J Cell Physiol，2006，290（4）：1139-1142.
［13］ POTIER E，FERREIRA E，ANDRIANMANALIJAONA R，et al. Hypoxia affects mesenchymal stromal cellosteogennic differention and angiogenic factor expression ［J］. Bone，2007，40（40）：1078-1084.
［14］ KOMORI T. Runx2，a multifunctional transcription factor in skeletal development ［J］. J Cell Biochem，2002，87（1）：1-8.
［15］ KEN O，LINDA J S. Extracellular matrix gene regulation ［J］. Clin Orthop，2004，427（5）：123-128.
［16］ HU Z，PEEL S A，HO S K，et al. Role of bovine bone morphogenetic proteins in bone matrix protein and osteoblast-related gene expression during rat bone marrow stem cell differentiation ［J］. J Craniofac Surg，2005，16（6）：1006-1014.
{{custom_fnGroup.title_cn}}
脚注
{{custom_fn.content}}
PDF(1623 KB)
