摘要
目的:研究微小RNA-23a(microRNA-23a,miR-23a)调控大鼠骨髓间充质干细胞(BMSCs)成软骨分化的作用和分子机制。方法:分离大鼠BMSCs并转染miR-23a基因后,通过RT-PCR和番红O染色法检测miR-23a调控大鼠BMSCs成软骨分化的作用,并进一步利用Western blot研究番红O调控BMSCs成软骨分化的分子机制。结果:转染miR-23a组和未转染组相比,成软骨分化相关基因如Sox9、二型胶原(CollagenⅡ)、聚集蛋白聚糖(Aggrecan)发生明显上调,而软骨肥大相关基因十型胶原(CollagenⅩ)表达发生下调。番红O染色显示转染miR-23a组和未转染组相比,硫酸软骨素表达亦发生明显上调。Western blot检测显示转染miR-23a能明显促进大鼠BMSCs的Sox9蛋白表达而抑制其Runx2蛋白表达。结论:miR-23a可能通过上调Sox9表达及抑制Runx2表达来达到促进BMSCs成软骨分化并抑制其进一步肥大的作用。
Objective：To study the effect and associated molecular mechanism of miR-23a on the chon- drogenic differentiation of rat bone marrow derived mesenchymal stem cells（BMSCs）. Methods： BMSCs were isolated from adult rats and then transfected with miR-23a. Chondrogenic differenti- ation associated genes were then analyzed by RT-PCR. Chondroitin sulfate expression was analyzed by sarranine O staining. Sox9 and Run2 protein expressions were evaluated by Western blot analysis. Results：BMSCs transfected with miR-23a showed a higher level of chondrogenic differentiation related genes such as Sox9, Collagen [[ and Aggreean. The expression of chondrocyte hypertrophy related genes Collagen X was lower in the miR-23a transfected group than the con- trol group. Chondroitin sulfate expression was also elevated in the miR-23a transfeeted group than the control group as shown by sarranine O staining. Finally, Sox9 protein expression wasincreased in the miR-23a transfected group and Runx2 protein expression was decreased in the miR-23a transfected group as demonstrated by Western blot analysis. Conclusion. miR-23a promotes chondrogenic differentiation and inhibits hypertrophy of BMSCs by increasing Sox9 expression and decreasing Runx2 expression.
出处
《武汉大学学报（医学版）》
CAS
北大核心
2014年第4期493-497,共5页
Medical Journal of Wuhan University
关键词
微小RNA-23a
骨髓间充质干细胞
成软骨分化
肥大
MieroRNA-23a
Bone Marrow Derived Mesenehymal Stem Cells
Chondrogenic Differentiation
Hyperthophy
