人脐血间质干细胞条件培养基在受损成骨细胞中的作用与机制

目的探讨线粒体活性在人脐带间充质干细胞(human umbilical cord mesenchymal stem cells,HUMSCs)条件培养基修复受损成骨细胞中的作用及其机制。方法提取、培养HUMSCs及制作其条件培养基,建立成骨细胞氧化损伤模型。实验分组:无血清DMEM培养基组、晚期氧化蛋白(advanced oxidation protein products,AOPPs)刺激组、常规培养基组、条件培养基组,培养1 h后用流式细胞仪以及Mito Tracker Red分析成骨细胞凋亡率、线粒体活性,并用蛋白-凝胶成像法(western-blot,WB)分析Caspase-3凋亡蛋白表达差异,从而评价HUMSCs条件培养基对遭受氧化损伤的成骨细胞的保护、修复、逆转作用,以及线粒体在其中发挥的作用及其机制。结果HUMSCs条件培养基可以增加氧化应激作用下成骨细胞内线粒体活性;HUMSCs条件培养基可降低成骨细胞凋亡率,减少Caspase-3凋亡蛋白表达。结论HUMSCs条件培养基通过缓解氧化应激(oxidative stress,OS)导致的线粒体功能与结构损伤,并下调Caspase-3凋亡蛋白的表达,抑制OB凋亡。

Role and mechanism of human umbilical cord blood mesenchymal stem cell conditioned medium in damaged osteoblasts

Objective To explore the role of mitochondrial activity in the repair of damaged osteoblasts with human umbilical cord mesenchymal stem cells (HUMSCs) conditioned medium and its mechanism. Methods The model of oxidative injury of osteoblasts was established. HUMSCs were cultured and the conditioned medium was extracted. Experimental groups included serum-free DMEM medium group, advanced oxidation protein products (AOPPs) stimulation group, conventional medium group, and conditioned medium group. After 1 hour of culture, the apoptosis rate and mitochondrial activity of osteoblasts were analyzed using flow cytometry and Mito Tracker Red. The difference of caspase-3 apoptotic protein expression was analyzed with protein-gel imaging (Western-Blotting, WB). The protective, repair, and reversal effect of HUMSCs conditioned medium on osteoblasts injured by oxidation, as well as the role and mechanism of mitochondriaIn, was evaluated. Results HUMSCs conditioned medium increased the activity of mitochondria in osteoblasts under oxidative stress and reduced the apoptosis rate of osteoblasts and the expression of apoptotic protein Caspase-3. Conclusion HUMSCs conditioned medium inhibits the apoptosis of osteoblasts by alleviating the damage of mitochondrial function and structure caused by oxidative stress and down-regulates the expression of apoptotic protein Caspase-3 .