Donor pretreatment with adenosine monophosphate-activated protein kinase activator protects cardiac grafts from cold ischaemia/reperfusion injury.
Donor pretreatment with adenosine monophosphate-activated protein kinase activator protects cardiac grafts from cold ischaemia/reperfusion injury.
复制标题
使用单磷酸腺苷激活蛋白激酶激活剂对供体进行预处理可保护心脏移植物免受冷缺血/再灌注损伤。
DOI：
10.1093/ejcts/ezv413
复制
发表时间：
2016
期刊：
影响因子：
--
通讯作者：
Z. Chen
中科院分区：
文献类型：
--
作者：
Chao Yang;honglai xu;L. Cai;Xiao;Yinan Jiang;Yong Zhang;Hongmin Zhou;Z. Chen
目标
腺苷一磷酸活化蛋白激酶（AMPK）是能量代谢的主要调节因子，在缺血/再灌注损伤（IRI）中具有保护作用。我们假设在供体心脏获取前用激活剂预激活AMPK可以保护心脏移植物免受冷IRI。
方法
供体Sprague-Dawley大鼠在心脏获取前30 min静脉注射AMPK激活剂5-氨基-咪唑-4-甲酰胺核糖核苷酸（AICAR）或溶媒。然后将心脏移植物在组氨酸-色氨酸-酮戊二酸（HTK）溶液中在4 ° C下保存8小时。保存后，立即将移植物固定在Langendorff灌注系统上，并在37 ° C下用Krebs-Henseleit缓冲液灌注1小时。保存后及再灌注后测定移植组织中三磷酸腺苷（ATP）和丙二醛（MDA）含量。再灌注后，使用压力传感器评估离体心脏功能;测量灌注液中肌酸激酶（CK）和乳酸脱氢酶（LDH）的累积释放以评估心肌细胞坏死;使用透射电子显微镜（TEM）检查移植物线粒体的超微结构变化。
结果
保存后，预处理组心肌ATP含量显著高于对照组（3.247 ± 0.3034 vs 1.817 ± 0.2533 μ mol/g蛋白; P <0.05）。AICAR预处理的心脏移植物在再灌注后表现出显著高于溶剂处理的冠状动脉流量（9.667 ± 0.3159 vs 8.033 ± 0.2459 ml/min; P <0.05）和左心室发展压（58.67 ± 2.894 vs 42.67 ± 3.333 mmHg; P <0.05）。AICAR预处理组灌流液中CK（300.0 ± 25.30 vs 431.7 ± 42.39 U/l; P <0.05）和LDH（228.0 ± 16.68 vs 366.8 ± 57.41 U/l; P <0.05）的累积释放量显著低于对照组。预处理组心肌MDA含量也明显降低（0.5167 ± 0.1046vs0.9333 ± 0.1333nmol/mg蛋白; P <0.05）。透射电镜表明，AICAR预处理的心脏的线粒体结构保存得更好。此外，AICAR预处理的心脏显着减少再灌注后细胞色素c的释放。
结论
这项研究表明，AMPK激活剂AICAR预处理可显著保护心脏移植物免受延长的冷IRI。这种新的方案可能是有用的和可行的，在临床心脏移植。
OBJECTIVES
Adenosine monophosphate-activated protein kinase (AMPK) is a master regulator of energy metabolism and has been shown to be protective in ischaemia/reperfusion injury (IRI). We hypothesized that preactivation of AMPK with an activator before donor heart procurement could protect heart grafts from cold IRI.
METHODS
Donor Sprague-Dawley rats were injected intravenously with AMPK activator 5-amino-imidazole-4-carboxamide ribonucleotide (AICAR) or vehicle 30 min before heart procurement. Heart grafts were then preserved in histidine-tryptophan-ketoglutarate (HTK) solution at 4°C for 8 h. After preservation, grafts were immediately mounted on the Langendorff perfusion system and perfused with Krebs-Henseleit buffer at 37°C for 1 h. Adenosine triphosphate (ATP) and malondialdehyde (MDA) content in graft tissue were quantified post-preservation and post-reperfusion. After reperfusion, isolated heart function was assessed using a pressure transducer; cumulative release of creatine kinase (CK) and lactate dehydrogenase (LDH) into the perfusate was measured to assess cardiomyocyte necrosis; ultrastructural changes in the mitochondria of the grafts were examined using transmission electron microscopy (TEM).
RESULTS
After preservation, myocardial ATP content in the pretreated hearts was significantly higher than in the control hearts (3.247 ± 0.3034 vs 1.817 ± 0.2533 µmol/g protein; P < 0.05). AICAR-pretreated heart grafts exhibited significantly higher coronary flow (9.667 ± 0.3159 vs 8.033 ± 0.2459 ml/min; P < 0.05) and left ventricular developing pressure (58.67 ± 2.894 vs 42.67 ± 3.333 mmHg; P < 0.05) than the vehicle treated after reperfusion. Cumulative release of CK (300.0 ± 25.30 vs 431.7 ± 42.39 U/l; P < 0.05) and LDH (228.0 ± 16.68 vs 366.8 ± 57.41 U/l; P < 0.05) in the perfusate was significantly lower in the AICAR-pretreated group than that in the control group. Myocardial MDA content was also reduced in the pretreated group (0.5167 ± 0.1046 vs 0.9333 ± 0.1333 nmol/mg protein; P < 0.05). TEM suggested that the mitochondrial structure of AICAR-pretreated hearts was much better preserved. Moreover, AICAR-pretreated hearts significantly diminished cytosolic cytochrome c release after reperfusion.
CONCLUSIONS
This study demonstrates that pretreatment with AMPK activator AICAR significantly protects heart grafts from extended cold IRI. This novel protocol may be useful and feasible in clinical heart transplantation.
影响因子：
--
作者：
ROUSLIN, W
通讯作者：
ROUSLIN, W
影响因子：
5
作者：
Kim AS;Miller EJ;Wright TM;Li J;Qi D;Atsina K;Zaha V;Sakamoto K;Young LH
通讯作者：
Young LH