摘要
研究淫羊藿素在GDF-5诱导BMSCs成软骨分化过程中的作用。全骨髓贴壁法分离培养SD大鼠骨髓间充质干细胞(BMSCs),取P3代细胞随机分成4组:对照组,淫羊藿素(Icaritin)组,Growth differentiation factor 5(GDF-5)组,Icaritin+GDF-5联合组。连续诱导培养14 d,倒置相差显微镜观察细胞形态,Alcian Blue染色检测细胞的蛋白聚糖改变,RT-PCR检测软骨分化标记基因Aggrecan、COL2、Sox9及COL1的表达情况,Western Blot检测COL2和COL1蛋白表达水平。结果提示,与对照组及GDF-5组相比,Icaritin+GDF-5联合组蛋白聚糖染色更深;软骨分化标记基因Aggrecan、COL2、Sox9明显增加;Ⅱ型胶原蛋白表达量均明显增加。淫羊藿素能够促进GDF-5诱导BMSCs成软骨分化。
In this study, the effects of iearitin on ehondregenic differentiation of bone marrow mesenehymal stem cells in- duced by growth differentiation factor 5 were investigated. BMSCs were collected from Sprague Dawley rats. The morphol- ogy was observed by inverted microscope, rBMSCs at passage 3 were randomized divided into 4 groups：control group, Icaritin group,GDF-5 group,Iearitin ＋ GDF-5 group and induced for 14 days. Aleian Blue stain showed the changes of preteoglycans ; RT-PCR was used to detect the mRNA expression of Aggrecan, COL2, Sox9 and COL1 ;the protein expres- sion levels of COL2 and COL1 were detected by Western Blot assay. The results indicated that, compared with the control group and GDF-5 group, Icaritin ＋ GDF-5 group showed deeper proteoglyeans staining; cartilage differentiation marker genes Aggrecan, COL2, Sox9 and the protein expression levels of COL2 significantly increased. Iearltin can promote chondrogenle differentiation of bone marrow mesenchymal stem ceils induced by GDF-5.
出处
《天然产物研究与开发》
CAS
CSCD
北大核心
2015年第11期1866-1870,1886,共6页
Natural Product Research and Development
基金
国家自然科学基金(81160226)
江西省教育厅基金(GJJ13740)
