摘要
目的研究白细胞蛋白酶抑制因子（secretory leukocyte protease inhibitor，SLPI）对真皮间充质干细胞（dermal mesenchymal stem cells，dMSCs）的增殖作用及其机制。方法利用贴壁生长的特性分离dMSCs，细胞在无血清IMDM培养液中培养48h后，加入不同浓度的SLPI，24h后加入[H^3]Thymidine，通过液闪计数器计数。提取对照组和SLPI作用后细胞的总RNA和蛋白，反转录-聚合酶链反应（RT—PCR）检测SLPI因子作用前后cyclin A、CDK2、CDK4的变化，Western blot检测SLPI因子作用前后cyclin D1、cyclin E的表达变化。结果分离的大鼠dMSCs呈梭形．在体外显示多向分化潜能。加入SLPI因子后，[H^3]Thymidine掺人活性增强。RT—PCR提示在SLPI诱导下，CDK2表达增强，而cyclin A和CDK4表达没有明显变化，Western blot提示SLPI因子作用后cyclin DI、cyclin E表达均明显增强。结论SLPI能够促进dMSCs的增殖，其机制可能是通过调节cychn D1、cyclin E、CDK2等细胞周期素的表达。
Objective To investigate the role of SLPI （secretory leukocyte protease inhibitor） in promoting proliferation of dMSCs as well as related mechanisms. Methods dMSCs were isolated from the rat skin tissues and cultured in serum-free IMDM medium for 48 hours. Then the cells were labeled with[ H^3 ] thymidine and the radioactivity was measured by automatic liquid scintillation counter. After the stimulation by SLPI, the expressions of cyclin A, cdk2 and cdk4 in dMSCs were determined by RT-PCR and the expressions of cyclin D1 and cyclinE were investigated using western-blot analysis. Results Most of the neonatal dMSCs were spindle-shaped cells and exhibited multilineage potential in vitro. A concentration-dependent proliferation promotion was demonstrated in dMSCs incubated with SLPI, compared with the control groups. The results from RT-PCR experiment indicated that this stimulation by SLPI increased the expression of cdk2, but had no effects on the expressions of cyclinA and cdk4. Mean- while, the western-blot analysis showed the increase of the expressions of cyclinD and cyclin E at the level of protein after SLPI stimulation. Conclusion SLPI can support the growth of dMSCs in serum-free medium in vitro. The molecular mechanism underlying this function may relate to the modulation of cell cycle progression via regulating cyclin DI, cyclin E and cdk2 expressions.
出处
《中华创伤杂志》
CAS
CSCD
北大核心
2007年第3期235-238,共4页
Chinese Journal of Trauma
基金
国家自然科学基金项目资助（30370562）
国家重点基础研究发展规划资助项目（2005CB522605）
关键词
干细胞
白细胞蛋白酶抑制因子
增殖
Stem ceils
Secretory leukocyte protease inhibitor
Proliferation
