摘要
目的:研究淫羊藿苷对大鼠骨髓间充质干细胞(rBMSCs)成骨和成脂方向分化的影响。方法:在37℃5%CO2和饱和湿度的条件下贴壁筛选法体外培养rBMSCs。采用1×10-5mol.L-1的淫羊藿苷干预rBMSCs的成骨性分化,第3,6,9,12天进行碱性磷酸酶(ALP)活性测定,第15天进行钙化结节染色,第12,24,48,72 h提取总RNA,采用实时聚合酶链反应(Real time PCR)方法分析检测骨成型蛋白2(BMP-2)和骨特异性转录因子(Runx-2)mRNA表达水平。采用1×10-5mol.L-1的淫羊藿苷干预rBMSCs的成脂性分化,第5,10,15,20天进行甘油三酯含量的测定,第21天进行油红O染色,于12,24,48,72 h提取总RNA,采用Real Time PCR方法分析检测PPARγ和C/EBPα的mRNA表达水平。结果:淫羊藿苷显著提高了细胞中ALP的活性,增加了钙化结节数,并且升高了成骨性相关因子BMP-2和Runx-2的基因表达量。淫羊藿苷降低了细胞中甘油三酯的含量,减少了脂滴的形成,并且减弱了成脂转录因子过氧化物酶体增生物激活受体γ(PPARγ)和增强子结合蛋白(C/EBPα)的基因表达量。结论:淫羊藿苷对rBMSCs分化的调节有双向性,即它在促进rBMSCs成骨性分化的同时又抑制rBMSCs成脂分化。
Objective： To investigate the effect of icariin on osteogenic differentiation and adipogenic differentiation of rat bone marrow stromal stem cells （rBMSCs）. Method： rBMSCs were incubated with final concentrations of 1 ~ i0-5 mol ~ L-1 icariin under the conditions of osteogenic induction culture. The alkalin phosphatase （ALP） activity was determined at day of 3,6, 9, 12 days and mineralized nodules were stained at the 15th days; Total RNA was isolated and the gene expressions of bone morphogenetic protein 2 （BMP-2） and Runx- 2 were investgated by Real time PCR. rBMSCs adipogenic differentiation were treatment by icariin with the concentration 1 x 10-Smol-L-1, triglycerides content was determined at day of 5, 10, 15 and 20, rathonum red staining was made at day of 21. Total RNA was isolated at 12, 24, 48 and 72 h to investigate the gene expression of PPAR and C/EBP mRNA by Real Time PCR. Result： Icariin significantly enhanced the ALP activity of the rBMSCs and the number of calcified nodules, and increased the gene expression of osteogenic factor BMP-2 and Runx-2. Icariin significantly reduced triglyceride levels and the formation of expression levels of the adipogenic transcription factors PPAR~ and C/EBPa rBMSCs differentiation, and promote rBMSCs osteogenic differentiation and suppression of rBMSCs adipogenic differentiation. lipid droplets, and decreased gene Conclusion： Icariin can regulate at the same time icariin can exert
出处
《中国实验方剂学杂志》
CAS
北大核心
2012年第20期200-204,共5页
Chinese Journal of Experimental Traditional Medical Formulae
基金
甘肃省自然科学研究基金(1107RJZA125)
甘肃省科技重大专项计划资助项目(09ZNKDA025)
关键词
淫羊藿苷
骨髓间充质干细胞
成骨分化
成脂分化
icariin
bone marrow stromal stem cells
osteogenic differentiation
adipogenic differentiation
