Wallace H. Coulter Department of Biomedical Engineering, Emory University and Georgia Institute of Technology, Atlanta, GA 30322, USA.
Stem Cells Dev. 2012 Nov 20;21(17):3136-46. doi: 10.1089/scd.2012.0191. Epub 2012 Aug 1.
Transplantation of cardiac progenitor cells (CPCs) is currently in early clinical testing as a potential therapeutic strategy. Superoxide is increased in the ischemic myocardium and poor survival of cells is one of the major limitations of cell transplantation therapy. Superoxide dismutase (SOD) levels were analyzed in c-kit-positive CPCs isolated from rat myocardium to identify their roles in protection against oxidative stress-induced apoptosis in vitro. CPCs were subjected to oxidative stress using xanthine/xanthine oxidase (XXO) and little apoptosis was detected. CPCs contained significantly higher levels of SOD1 and SOD2 as compared with adult cardiac cell types, both at the protein and activity levels. Both SOD1 and SOD2 were increased by XXO at the mRNA and protein level, suggesting compensatory adaptation. Only knockdown of SOD2 and not SOD1 with siRNA sensitized the cells to XXO-apoptosis, despite only accounting for 10% of total SOD levels. Finally, we found XXO activated Akt within 10 min, and this regulated both SOD2 gene expression and protection against apoptosis. Rat CPCs are resistant to superoxide-induced cell death, primarily through higher levels of SOD2 compared to adult cardiac-derived cells. Exposure to superoxide increases expression of SOD2 in an Akt-dependent manner and regulates CPC survival during oxidative stress.
心脏祖细胞(CPCs)的移植目前处于临床前期试验阶段,作为一种潜在的治疗策略。在缺血性心肌中,超氧化物增加,细胞存活率低是细胞移植治疗的主要限制之一。为了确定其在体外对抗氧化应激诱导的细胞凋亡中的作用,分析了从大鼠心肌中分离出的 c-kit 阳性 CPCs 中的超氧化物歧化酶(SOD)水平。用黄嘌呤/黄嘌呤氧化酶(XXO)使 CPCs 产生氧化应激,检测到很少的细胞凋亡。与成人心肌细胞类型相比,CPCs 在蛋白质和活性水平上均具有显著更高水平的 SOD1 和 SOD2。XXO 可在 mRNA 和蛋白质水平上增加 SOD1 和 SOD2 的表达,表明代偿性适应。尽管仅占总 SOD 水平的 10%,但只有 SOD2 的 siRNA 敲低而不是 SOD1 的敲低使细胞对 XXO-凋亡敏感。最后,我们发现 XXO 在 10 分钟内激活 Akt,这调节了 SOD2 基因表达和对细胞凋亡的保护。大鼠 CPCs 对超氧化物诱导的细胞死亡具有抗性,主要是通过与成人心肌衍生细胞相比具有更高水平的 SOD2。超氧化物暴露以 Akt 依赖性方式增加 SOD2 的表达,并调节 CPC 在氧化应激期间的存活。