摘要
[目的]探究5-氮杂胞苷和地塞米松对大鼠骨髓间充质干细胞(mesenchymal stem cells,MSCs)成骨分化能力的影响。[方法]SD大鼠骨髓间充质干细胞体外扩增和传代培养,流式细胞术鉴定细胞表面标志,取3代细胞随机分为空白组、激素组、激素+5-氮杂胞苷组,相应药物干预3 d后成骨诱导分化14 d,RT-PCR和Western blot检测Runx2和PPARγ的表达。[结果]所取得骨髓间充质干细胞纯度和均一性好,细胞表面标记CD90高表达,CD34、CD45低表达。第3代细胞经药物干预和成骨诱导后,与空白组相比,激素组Runx2基因表达降低,PPARγ基因升高,5-氮杂胞苷+激素组结果与激素组相反(P<0.05)。[结论]激素能抑制骨髓间充质干细胞成骨分化,促进成脂分化,5-氮杂胞苷能干预激素的这种作用,提高骨髓间充质干细胞分化潜能。
[Objective] To investigate the effects of 5-azacytidine and dexamethasone on osteogenic differentiation of bone marrow mesenchymal stem cells （MSCs） in rats in vitro. [Methods] Bone marrow MSCs of Sprague-Dawley rats were cultured in vitro. The cell surface markers were analyzed by flow cytometry. The passage 3 ceils were randomly divided into three groups： those in the dexamethasone group were cultured in the medium with 10^-5M dexamethasone, and those in the 5-aza- cytidine＋dexamethasone group were cultured in the medium with 10^-5 M dexamethasone and 10 μM 5-azacytidine, meanwhile, the control group contained the cells cultured in the normal medium. After 72-h treatment, the osteogenic differentiation was in- duced for all three groups by culturing MSCs in osteogenic medium for 14 days. And then the mRNA and protein expression of Runt-related transcription factor 2 （Runx2） and peroxisome proliferator-activated receptor γ （PPARγ） was measured by RT-PCR and Western blot, respectively. [Results] The cultured MSCs had good purity and homogeneity with high expres- sion of CD90 and low expression of CD34 and CD45 on cell surfaces. After drug intervention and osteogenic differentiation, com- pared with the control group, the dexamethasone group had significantly lower mRNA expression of Runx2 and significantly higher mRNA expression of PPARγ （P〈0.05） , while the 5-azacytidine ＋ dexamethasone group had significantly higher mRNA expression of Runx2 and significantly lower mRNA expression of PPARγ （P〈0.05） . [Conclusions] Dexamethasone can inhibit the osteogenic differentiation of bone marrow MSCs, while 5-azacytidine can promote the osteogenic differentiation of bone marrow MSCs.
出处
《中国矫形外科杂志》
CAS
CSCD
北大核心
2017年第1期58-63,共6页
Orthopedic Journal of China
基金
国家自然科学基金资助项目(编号:81271969)
关键词
5-氮杂胞苷
骨髓间充质干细胞
糖皮质激素
成骨分化
5-Azacytidine, bone marrow mesenchymal stem cell, dexamethasone, osteogenic differentiation
