中文摘要：

目的龟板有促大鼠骨髓间充质干细胞（MSCs）增殖作用,本研究观察龟板含药血清对MSCs中骨形态发生蛋白4（BMP4）表达的影响。方法使用密度梯度法分离大鼠骨髓间充质干细胞进行培养,传代纯化,在基础培养液中添加不同浓度龟板含药血清和对照血清,用荧光定量PCR和原位杂交方法检测BMP4 mRNA表达,用ELISA、免疫组织化学和免疫荧光组织化学法结合激光扫描共聚焦显微镜技术检测BMP4蛋白表达的变化。结果BMP4蛋白和mRNA表达相一致,龟板含药血清以浓度依赖方式促进MSC的BMP4 mRNA及其蛋白表达。结论龟板含药血清以时效和量效依赖性促进BMP4在MSCs的表达,可能与龟板促MSCs增殖作用有关。

英文摘要：

Objective To observe the effect of seropharmacological plastrum testudinis on the expression of bone morphogenetic protein4 （BMP4） of rat mesenchymal stem cells （MSCs） in vitro. Methods Mesenchymal stem cells were dissociated from rat bone marrow and were marked by Brdu and the expression of CD44. The growth of rat mesenchymal stem cells with low, middle and high seropharmacological plastrum testudinis were observed separately by means of morphological apperances. The locations of the BMP4 protein and BMP4 mRNA positive cells were detested by immunohistochemistry and fluorescent immunohistochemistry combined with laser confocal scanning microscopy and in situ hybridization stained by DAB. The expression quantity of BMP4 mRNA and BMP4 protein was detected by real time PCR and ELISA. Results Seropharmacological plastrum testudinis promoted the numbers of BMP4 and BMP4 mRNA positive cells and the expressions of BMP4 protein and BMP4 mRNA in a time- and dose-dependant manner; there were significant differences in comparison with control groups. Conclusion Promoting effect of seropharmacological plastrum testudinis on the proliferation of MSC might be related to the enhanced expression of BMP4.