肾上腺髓质素对缺氧培养骨髓间充质干细胞表达凋亡相关蛋白的影响
发布时间：2018-12-30 12:34
【摘要】：背景:肾上腺髓质素基因转染能增强骨髓间充质干细胞在缺血、缺氧环境下的抗凋亡能力,但其机制尚未完全明确。目的:观察肾上腺髓质素对骨髓间充质干细胞缺血、缺氧时Bax、Bcl-2和Caspase-3等凋亡相关调节蛋白表达的影响。方法:贴壁法分离、培养、纯化SD大鼠骨髓间充质干细胞,缺氧、无血清培养0,3,6,9,12 h后用Western blot测定Bax、Bcl-2、Caspase-3蛋白的表达以确定最佳缺氧时间。采用缺氧6 h为时间点,根据缺氧、无血清培养前是否进行肾上腺髓质素预处理分为对照组(未加入肾上腺髓质素)及肾上腺髓质素不同质量浓度组(1,10,100μg/L),然后采用Western blot测定Bax、Bcl-2、Caspase-3蛋白的表达。结果与结论:1无血清培养条件下缺氧0,3,6,9,12 h后骨髓间充质干细胞Bax、Bcl-2表达均增加(P0.05),缺氧6 h时Bax/Bcl-2比值、caspase-3蛋白达到最小(P0.05)。2缺氧6 h时,与1,10μg/L肾上腺髓质素组相比,100μg/L肾上腺髓质素组表达Bax蛋白、Caspase-3蛋白最低,Bax/Bcl-2比值最低,而表达Bcl-2蛋白最高(P0.05)。结果表明肾上腺髓质素能降低无血清、缺氧条件下骨髓间充质干细胞Bax/Bcl-2比值、Caspase-3蛋白表达,该效应存在剂量依赖性。
[Abstract]:Background: adrenomedullin gene transfection can enhance the anti-apoptosis ability of bone marrow mesenchymal stem cells in ischemic and hypoxic environment, but its mechanism is not completely clear. Aim: to investigate the effects of adrenomedullin on the expression of apoptosis-related regulatory proteins such as Bax,Bcl-2 and Caspase-3 during ischemia and hypoxia of bone marrow mesenchymal stem cells. Methods: bone marrow mesenchymal stem cells (BMSCs) of SD rats were isolated, cultured and purified by adherent method. After hypoxia and serum-free culture for 12 h, the expression of Bax,Bcl-2,Caspase-3 protein was determined by Western blot to determine the best hypoxia time. According to whether adrenomedullin pretreatment was performed before anoxia and serum-free culture for 6 h, the rats were divided into two groups: control group (without adrenomedullin) and adrenomedullin with different concentration (1 10 100 渭 g / L). Then Western blot was used to detect the expression of Bax,Bcl-2,Caspase-3 protein. Results and conclusion: 1 the expression of Bax,Bcl-2 in bone marrow mesenchymal stem cells increased after 12 h hypoxia (P0.05), and the Bax/Bcl-2 ratio increased at 6 h after hypoxia. Caspase-3 protein reached the minimum (P0.05). 2 at 6 h hypoxia, compared with 1 渭 g / L adrenomedullin group, 100 渭 g / L adrenomedullin group expressed Bax protein, Caspase-3 protein was the lowest, and Bax/Bcl-2 ratio was the lowest. The expression of Bcl-2 protein was the highest (P0.05). The results showed that adrenomedullin could decrease the ratio of Bax/Bcl-2 and the expression of Caspase-3 protein in cultured bone marrow mesenchymal stem cells in a dose-dependent manner.
【作者单位】： 福建省立医院北院福建省老年医院心内科;福建医科大学附属第一医院心内科高血压研究所;
【基金】：福建省卫生厅青年基金(2011-1-15,2011-1-32) 福建省自然科学基金(2014J01433)~~
【分类号】：R329.2
[Abstract]:Background: adrenomedullin gene transfection can enhance the anti-apoptosis ability of bone marrow mesenchymal stem cells in ischemic and hypoxic environment, but its mechanism is not completely clear. Aim: to investigate the effects of adrenomedullin on the expression of apoptosis-related regulatory proteins such as Bax,Bcl-2 and Caspase-3 during ischemia and hypoxia of bone marrow mesenchymal stem cells. Methods: bone marrow mesenchymal stem cells (BMSCs) of SD rats were isolated, cultured and purified by adherent method. After hypoxia and serum-free culture for 12 h, the expression of Bax,Bcl-2,Caspase-3 protein was determined by Western blot to determine the best hypoxia time. According to whether adrenomedullin pretreatment was performed before anoxia and serum-free culture for 6 h, the rats were divided into two groups: control group (without adrenomedullin) and adrenomedullin with different concentration (1 10 100 渭 g / L). Then Western blot was used to detect the expression of Bax,Bcl-2,Caspase-3 protein. Results and conclusion: 1 the expression of Bax,Bcl-2 in bone marrow mesenchymal stem cells increased after 12 h hypoxia (P0.05), and the Bax/Bcl-2 ratio increased at 6 h after hypoxia. Caspase-3 protein reached the minimum (P0.05). 2 at 6 h hypoxia, compared with 1 渭 g / L adrenomedullin group, 100 渭 g / L adrenomedullin group expressed Bax protein, Caspase-3 protein was the lowest, and Bax/Bcl-2 ratio was the lowest. The expression of Bcl-2 protein was the highest (P0.05). The results showed that adrenomedullin could decrease the ratio of Bax/Bcl-2 and the expression of Caspase-3 protein in cultured bone marrow mesenchymal stem cells in a dose-dependent manner.
【作者单位】： 福建省立医院北院福建省老年医院心内科;福建医科大学附属第一医院心内科高血压研究所;
【基金】：福建省卫生厅青年基金(2011-1-15,2011-1-32) 福建省自然科学基金(2014J01433)~~
【分类号】：R329.2
【参考文献】
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