低氧预处理通过激活AKT通路提高老年hBM-MSCs对氧化应激损伤的耐受能力

发布时间：2018-06-05 11:50

本文选题：低氧 + PIK/AKT信号通路　；参考：《中国病理生理杂志》2016年05期

【摘要】：目的:探讨低氧预处理对老年人骨髓间充质干细胞(h BM-MSCs)的保护作用,为提高老年自体干细胞移植治疗效果提供实验支持。方法:老年h BM-MSCs于低氧培养箱中培养24 h进行低氧预处理,实验分为年轻h BM-MSCs组(young组),老年h BM-MSCs组(old组)及低氧预处理老年h BM-MSCs组(old+hypoxia组)。300μmol/L H_2O_2作用30 min建立细胞氧化应激模型,50μmol/L LY294002作用2 h阻断PI3K/AKT信号通路,Brd U掺入实验检测细胞增殖能力;CCK-8法检测细胞活力,Western blot检测凋亡相关蛋白Bax、Bcl-2表达水平和AKT磷酸化水平。结果:Brd U掺入实验显示低氧预处理的老年h BM-MSCs细胞阳性率为39.85%±3.45%,与old组相比增殖能力显著提高(P0.05)。300μmol/L H_2O_2作用30 min诱导细胞氧化应激后,old+hypoxia组与old组比较,细胞活力显著提高(P0.05),凋亡相关蛋白Bax表达量显著降低(P0.05),抑制凋亡的Bcl-2蛋白表达量显著增高(P0.05),且AKT磷酸化水平显著增高,差异有统计学显著性(P0.05);应用LY294002抑制PI3K/AKT信号通路后,细胞活力下降(P0.05)。结论:低氧预处理可以通过激活AKT信号通路提高老年人骨髓间充质干细胞活力及增殖能力。
[Abstract]:Objective: to investigate the protective effect of hypoxic preconditioning on bone marrow mesenchymal stem cells (BM-MSCs) in the elderly and to provide experimental support for the treatment of autologous stem cell transplantation. Methods: the aged BM-MSCs was cultured in hypoxia incubator for 24 hours. The experiment was divided into three groups: young group in young BM-MSCs group, BM-MSCs group in old age group (n = 30 min) and hypoxia preconditioning group in old BM-MSCs group (n = 30 min). The model of cell oxidative stress was induced by 50 渭 mol/L LY294002 for 2 h to block the signal pathway of PI3K/AKT. The cell viability was detected by CCK-8 method. The expression of apoptosis related protein Baxfen Bcl-2 and the level of AKT phosphorylation were detected by Western blot. Results the positive rate of BM-MSCs cells was 39.85% 卤3.45% after hypoxia preconditioning. Compared with old group, the proliferation ability of old group was significantly higher than that of old group. Compared with old group, the proliferation ability of hypoxia group was significantly higher than that of old group after 30 min induction of oxidative stress. The cell viability was increased significantly (P 0.05), the expression of apoptosis-related protein (Bax) was significantly decreased, the expression of Bcl-2 protein was significantly increased (P 0.05), and the level of AKT phosphorylation was significantly increased (P 0.05). The cell viability was decreased by P0.05. Conclusion: hypoxia preconditioning can enhance the activity and proliferation of bone marrow mesenchymal stem cells by activating AKT signaling pathway.
【作者单位】：山西医科大学人体解剖学教研室;山西医科大学形态学实验室;山西医科大学第二医院;
【基金】：山西医科大学青年基金资助项目(No.02201002);山西医科大学基础医学院331基金资助项目(No.201217)
【分类号】：R457.7

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