PDF(Pubmed)
Abstract：
Donation after circulatory death (DCD) is a promising strategy for alleviating donor shortage in heart transplantation. Trehalose, an autophagy inducer, has been shown to be cardioprotective in an ischemia-reperfusion (IR) model; however, its role in IR injury in DCD remains unknown. In the present study, we evaluated the effects of trehalose on cardiomyocyte viability and autophagy activation in a DCD model. In the DCD model, cardiomyocytes (H9C2) were exposed to 1 h warm ischemia, 1 h cold ischemia, and 1 h reperfusion. Trehalose was administered before cold ischemia (preconditioning), during cold ischemia, or during reperfusion. Cell viability was measured using the Cell Counting Kit-8 after treatment with trehalose. Autophagy activation was evaluated by measuring autophagy flux using an autophagy inhibitor, chloroquine, and microtubule-associated protein 1A/1B light chain 3 B (LC3)-II by western blotting. Trehalose administered before the ischemic period (trehalose preconditioning) increased cell viability. The protective effects of trehalose preconditioning on cell viability were negated by chloroquine treatment. Furthermore, trehalose preconditioning increased autophagy flux. Trehalose preconditioning increased cardiomyocyte viability through the activation of autophagy in a DCD model, which could be a promising strategy for the prevention of cardiomyocyte damage in DCD transplantation.
摘要：
循环性死亡（DCD）后捐赠是缓解心脏移植供体短缺的有希望的策略。海藻糖,自噬诱导剂,已被证明在缺血再灌注（IR）模型中具有心脏保护作用；然而，其在DCDIR损伤中的作用尚不清楚。在本研究中,我们在DCD模型中评价了海藻糖对心肌细胞活力和自噬激活的影响.在DCD模型中,心肌细胞（H9C2）暴露于热缺血1小时，冷缺血1h,再灌注1h。在冷缺血（预处理）之前给予海藻糖，在冷缺血期间，或再灌注期间。在用海藻糖处理后,使用细胞计数试剂盒-8测量细胞活力。通过使用自噬抑制剂测量自噬通量来评估自噬激活，氯喹,和微管相关蛋白1A/1B轻链3B（LC3）-II通过蛋白质印迹。在缺血期之前施用海藻糖(海藻糖预处理)增加细胞活力。氯喹处理否定了海藻糖预处理对细胞活力的保护作用。此外，海藻糖预处理增加自噬通量。海藻糖预处理通过激活DCD模型中的自噬增加心肌细胞的活力，这可能是预防DCD移植中心肌细胞损伤的有希望的策略。
