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  

Effect of hyperbaric oxygen on the bone marrow mesenchymal stem cells against hypoxia/reoxygenation induced injury

作　　者：张慧[1] 陈银海[2] 李萌[2] 黄玉琴[1] 

机构地区：[1]南方医科大学第二临床医学院,广州510515 [2]南方医科大学珠江医院高压氧科

出　　处：《中国康复医学杂志》2017年第2期167-173,共7页Chinese Journal of Rehabilitation Medicine

基　　金：广东省科技计划项目(2010B080701020)

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%CO_2密闭培养箱中培养9h,复氧6h)、缺氧/复氧+高压氧组C(90%O_2,2.5ATA、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。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% CO_2 for 9h, followed by 12 h of reoxygenation）, hypoxia/reoxygenation ＋ hyperbaric oxygen group C（90% O_2, 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 re-duce 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.

关 键 词：高压氧 缺氧/复氧 骨髓间充质干细胞 

分 类 号：R-332[医药卫生] R651.2