龟板含药血清对骨髓间充质干细胞增殖细胞核抗原表达的影响

目的:观察益肾龟板血清对大鼠骨髓间质干细胞(mesenchymal stem cells,MSC)体外增殖过程的增殖细胞核抗原(Proliferation cell nuclear antigen,PCNA)表达的影响。方法:使用密度梯度法分离大鼠骨髓间质干细胞进行培养,经Brdu标记和CD44染色及两者双重染色鉴定后,观察在培养液中添加不同浓度龟板血清条件下MSC增殖过程。用免疫组织化学方法和原位杂交方法检测MSC的PCNA及其mRNA表达;免疫荧光细胞化学法结合激光扫描共聚焦显微镜技术检测PCNA蛋白表达的变化。结果:龟板血清以浓度依赖方式促进MSC的PC-NA及其mRNA表达阳性细胞数和PCNA蛋白表达,与对照组比较有显著性(P<0.05)。结论:龟板血清促MSC增殖作用可能与上调PCNA表达有关。

Effects of Seropharmacological Plastrum Testudinis on Proliferation Cell Nuclear Antigen of Rat Mesenchymal Stem Cell in Vitro

Objective:To observe the effect of Seropharmacological Plastrum Testudinis on Proliferation cell unclear antigen(PCNA) of rat mesenchymal stem cell(MSC) 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 cell under Seropharmacological Plastrum Testudinis were observed by means of morphological observation.The change of the PCNA and PCNA mRNA positive cells was studied by immunohistochemistry and in situ hybridization.The expression of PCNA protein was detected by using immunofluoresence and laser confocal scanning microscopy.Results:Seropharmacological Plastrum Testudinis can promote the number of PCNA positive cell and the expression of PCNA protein in dose-dependant,there are significant difference in comparison with control groups.Conclusion:Promoting effect of Seropharmacological Plastrum Testudinis on proliferation of MSC might be relate to the up-regulated expression of PCNA.