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张 慧,陈银海,李 萌,黄玉琴.高压氧对缺氧/复氧模型下大鼠骨髓间充质干细胞的影响及其相关机制[J].中国康复医学杂志,2017,(2):167~173
高压氧对缺氧/复氧模型下大鼠骨髓间充质干细胞的影响及其相关机制    点此下载全文
张 慧  陈银海  李 萌  黄玉琴
南方医科大学第二临床医学院, 广州, 510515
基金项目:广东省科技计划项目(2010B080701020)
DOI:
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摘要:
      摘要 目的:探讨高压氧对缺氧/复氧模型下大鼠骨髓间充质干细胞的影响及其相关机制。 方法:选取3—4周龄SD雄性大鼠,采用全骨髓贴壁法培养至P3代,随机分为正常组A(细胞置于常规培养箱内培养)、缺氧/复氧组B(置于95%N2、5%CO2密闭培养箱中培养9h,复氧6h)、缺氧/复氧+高压氧组C(90%O2,2.5ATA、90min)、缺氧/复氧+高压氧+YC-1组D,实验组其余时间处理和正常组相同。A、B、C、D组均于处理结束12h后行CCK-8比色法、流式细胞仪检测细胞存活率及凋亡率。Western blot检测HIF-1α、β-catenin、LEF-1蛋白表达水平。 结果:①与A组相比,缺氧/复氧后B、C、D组细胞存活率明显降低,差异具有显著性意义(P<0.05),给予高压氧后C组存活率比B组升高,D组存活率较C组降低(P<0.05)。②实验组B、C相比,C组中HIF-1α、β-catenin、LEF-1蛋白表达水平升高(P<0.05)。③实验组C、D组相比,给予YC-1阻断剂后HIF-1α、β-catenin、LEF-1蛋白表达减少(P<0.05)。 结论:高压氧能够提高缺氧环境下骨髓间充质干细胞存活率,其机制可能与调控HIF-1α介导的Wnt通路有关。
关键词:高压氧  缺氧/复氧  骨髓间充质干细胞
Effect of hyperbaric oxygen on the bone marrow mesenchymal stem cells against hypoxia/reoxygenation induced injury    Download Fulltext
Zhujiang Hospital of Southern Medical University,510282
Fund Project:
Abstract:
      Abstract Objective: To investigate whether hyperbaric oxygen could protect the bone marrow mesenchymal stem cells against apoptosis. after hypoxia/reoxygenation induced injury. Method: MSCs were generated from the bone marrow of adult male SD rats aged 3—4 weeks, they were randomly divided into control group A (cells were cultured in a normoxia), hypoxia/reoxygenation group B (cells were treated with 95% N2, 5% CO2 for 9h, followed by 12h of reoxygenation), hypoxia/reoxygenation + hyperbaric oxygen group C (90% O2, 2.5ATA, 90min), hypoxia/reoxygenation + hyperbaric oxygen + YC-1 group D. Besides, the experimental group were treated the same as the normal group. 12 hours after treatment,the rate of cell survival and apoptosis in different groups was estimated by CCK-8 and flow cytometry analysis. HIF-1α, β-catenin, LEF-1 were detected by Western blotting. Result: ① Compared with group A, the rate of cell survival in group B, C, D were significantly decreased (P<0.05), while administration of hyperbaric oxygen, the survival in group C was higher than group B and this could be reversed by YC-1 in group D (P<0.05). ② As compared to group B, the expression of HIF-1α, β-catenin, LEF-1 were increased in group C (P<0.05). ③ Compared to group C, YC-1 in group D could reduce HIF-1α, β-catenin, LEF-1 (P<0.05). Conclusion: Hyperbaric oxygen produces protective effects on survival of BMSCs against hypoxia/reoxygenation injury and possibly mediated by HIF-1α in Wnt pathway.
Keywords:hyperbaric oxygen  hypoxia/reoxygenation  bone marrow mesenchymal stem cells
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