摘要
目的:观察老鹳草素对小鼠骨髓基质干细胞(BMSCs)向成骨细胞分化的影响,并对成骨分化过程中相关基因表达进行检测。方法:分离、培养小鼠BMSCs,加入不同浓度老鹳草素(1×10-9,1×10-8,1×10-7mol·L-1)培养,另设空白组,分别于成骨诱导第7,14天采用碱性磷酸酶(ALP),骨钙素(OCN)试剂盒测定ALP活性及OCN含量,并进行ALP染色。于成骨诱导第21天时采用茜素红染色检测成骨分化过程中钙结节形成数量,实时荧光定量聚合酶链式反应(Real-time PCR)和蛋白质免疫印迹(Western blot)检测BMSCs成骨分化过程中OCN,ALP,骨涎蛋白(BSP),Runt相关转录因子2(Runx2)mRNA和蛋白表达的变化。结果:与空白组比较,成骨诱导后第7,14天时,老鹳草素各剂量组ALP活性,OCN含量及ALP染色强度均明显升高(P<0.05),成骨诱导第21天时,老鹳草素各剂量组矿化结节数量及OCN,ALP,BSP,Runx2 mRNA和蛋白表达均明显升高(P<0.05)。结论:老鹳草素能够明显促进BMSCs向成骨方向分化,成骨相关基因OCN,ALP,BSP,Runx2均参与这一过程。
Objective： To study the effects of geraniin on the osteogenic gene expression of murine bone marrow mesenchymal stem cells（BMSCs） during osteogenic induction. Method： BMSCs were isolated from mice bones,and cultured with different concentration of geraniin（1 × 10- 9,1 × 10- 8,1 × 10- 7mol·L- 1）. The blank group was also set up. After 7,14 days of osteogenic induction,the activity of alkaline phosphatase（ALP）,and osteocalcin（OCN） content were detected,and ALP staining was performed. Alizarin red staining was adopted to detect calcium deposits on 21 d. Real-time PCR and Western blot were applied to detect the changes in expressions of osteoblast differentiation marker genes OCN,ALP,bone sialoprotein（BSP）,Runx2. Result： At 7,14 d of osteogenic induction,geraniin groups showed significant increases in ALP activity,OCN content and ALP staining intensity,compared with control group（P 〈0. 05）. At 21 d of osteogenic induction,geraniin groups showed significant increases in the number of mineralized nodule,and the mRNA and protein expressions of OCN,ALP,BSP and Runx2（P〈 0. 05）. Conclusion： Geraniin could induce osteogenic differentiation of BMSCs. Osteoblast differentiation marker genes OCN,ALP,BSP and Runx2 participate in this process.
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2016年第22期140-144,共5页
Chinese Journal of Experimental Traditional Medical Formulae
基金
河北省中医药管理局科研计划项目(2015137)
关键词
老鹳草素
骨髓基质干细胞
成骨分化
geraniin
bone marrow mesenchymal stem cell
osteogenic differentiation
