【作者】 谭国庆；

【导师】 徐展望；

【作者基本信息】 山东中医药大学 ， 中医骨伤科学， 2005， 硕士

【摘要】 目的:观察骨碎补提取液能否促进体外培养的骨髓基质细胞(Bone Marrow Stromal Cells,BMSCs)向成骨细胞分化,初步探讨骨碎补促进骨折愈合、防治骨质疏松可能的一种细胞学机制。方法:用不同浓度的骨碎补醇提液对骨髓基质细胞持续培养,通过倒置相差显微镜、扫描电镜、组织化学染色、钙染色等手段对所培养细胞进行形态学和组织学观察,同时与标准培养液及成骨条件诱导培养液持续培养的细胞进行对比研究。结果:高浓度的药物对细胞产生了有害影响。含中、低浓度的药物所培养的部分细胞,在体外逐渐增殖、转化、分泌细胞外基质、最终能形成矿化基质;组织学鉴定显示碱性磷酸酶染色呈阳性,阳性率均在30%以上,并能大量分泌Ⅰ型胶原,持续培养能形成钙结节;同空白对照组和诱导对照组所培养细胞比较,从碱性磷酸酶染色阳性细胞数、Ⅰ型胶原表达及形成钙结节的能力来看,中、低浓度的中药组虽还远不及经典诱导剂的作用,但同空白对照组相比,已有了显著的影响,而两组之间无明显差异。结论:中药骨碎补的醇提取液能促进骨髓基质细胞向成骨细胞分化,对骨髓基质细胞的这种作用可能是其促进骨质愈合、预防治疗骨质疏松的一种细胞学机制。更多还原

【Abstract】 In the traditional Chinese medicine, Gu-Sui-Bu[Drynaria Fortunei(kunze) J. Sm] has been reported as a good enhancer for bone healing. In this experiment, we investigate the effects of this traditional Chinese medicine on osteogenic differentiation of bone marrow stromal cells in vitro. And try to discuss that marrow stromal cells is a kind of potential target cell of Gu-Sui-Bu [Drynaria Fortunei(kunze) J. Sm] or these traditional Chinese medicine that were proved to regulate kidney to promote bone qualities and enhance bone healing.Different concentrations of ethanol extract of Gu-Sui-Bu were added to rabbit marrow stromal cells culture. The effect on osteoblastic differentiation was examined by reserved phase-contrast microscope, scanning electron microscope, histochemistry stains and calcific nodules stains. In the same time, osteoblastic differentiation of these cells cultured by both a condition medium including dexamethasone, sodium β -glycerophosphate and vitamine C and pure medium were examined in same methods.The effective concentrations of Gu-Sui-Bu on osteoblastic differentiation of were 2mg/ml and 0.2mg/ml.The addition of 2mg/ml and 0.2mg/ml respectively to bone marrow stromal cells culture partly expressed osteoblastic phenotype.Firstly these cells induced began to differentiation or transform, then proliferated and secreted, ultimately came into being mineralized matrices in vitro. Histochemical studies更多还原