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索马(Somah)的进一步评估:长期保存、温度影响以及对心脏循环死亡后摘取的大鼠心脏缺血再灌注损伤的预防

Further evaluation of Somah: long-term preservation, temperature effect, and prevention of ischemia-reperfusion injury in rat hearts harvested after cardiocirculatory death.

作者信息

Lowalekar S K, Lu X G, Thatte H S

机构信息

Cardiothoracic Surgery Division, Department of Surgery, VA Boston Healthcare System, Boston, Massachusetts, USA; Brigham and Women's Hospital, Boston, Massachusetts, USA; Harvard Medical School, Boston, Massachusetts, USA.

出版信息

Transplant Proc. 2013 Nov;45(9):3192-7. doi: 10.1016/j.transproceed.2013.06.006.

Abstract

OBJECTIVE

To identify and evaluate the ideal temperature for long-term storage of hearts from donation after cardiocirculatory death, in the novel organ preservation solution Somah.

METHODS

DCD hearts from Sprague-Dawley rats were harvested after 30 minutes of euthanasia, preserved in Somah at 4°C, 10°C, 21°C, or 37°C for 24 hours and then reperfused with blood:Somah (3:1) perfusate at 37°C for 30 minutes. Myocardial biopsies were taken during storage and before and after reperfusion to assess the structural and functional viability of tissue using multiphoton imaging, biochemistry, and immunofluorescence.

RESULTS

Myocyte viability, determined by Live-Dead and esterase assays, was similar at 4°C, 10°C, and 21°C (193, 198 and 217 normalized fluorescence counts [NFC]) with a significant decrease at 37°C (131 NFC). Upon reperfusion, esterase activity was enhanced in DCD hearts stored in Somah at 21°C but noticeably decreased at all other temperatures. High-energy adenosine triphosphate/creatine phosphate (ATP/CP) syntheses and the expression of structural/contractile proteins was well preserved at 21°C, both after 24-hour storage and upon reperfusion. In contrast, hearts stored at all other temperatures demonstrated variable degenerative changes, loss of protein expression, and/or deranged ATP/CP synthesis after 24 hours of storage and/or upon reperfusion.

CONCLUSION

The robust maintenance of structural/functional integrity of cardiac tissue and the preservation of protein expression and cellular energy metabolism in DCD hearts after long-term preservation at subnormothermic temperature suggests that 21°C is ideal for long-term storage of DCD hearts in Somah solution.

摘要

目的

在新型器官保存溶液索马(Somah)中,确定并评估心循环死亡后供体心脏长期保存的理想温度。

方法

对安乐死30分钟后的斯普拉格-道利大鼠的心脏进行采集,将其在4℃、10℃、21℃或37℃的索马溶液中保存24小时,然后在37℃用血液:索马(3:1)灌注液再灌注30分钟。在保存期间以及再灌注前后进行心肌活检,使用多光子成像、生物化学和免疫荧光评估组织的结构和功能活力。

结果

通过活死检测和酯酶检测确定的心肌细胞活力在4℃、10℃和21℃时相似(分别为193、198和217归一化荧光计数[NFC]),在37℃时显著降低(131 NFC)。再灌注时,在21℃保存于索马溶液中的心循环死亡心脏的酯酶活性增强,但在所有其他温度下均明显降低。在24小时保存后以及再灌注后,21℃时高能三磷酸腺苷/磷酸肌酸(ATP/CP)合成以及结构/收缩蛋白的表达均得到良好保存。相比之下,在所有其他温度下保存的心脏在24小时保存后和/或再灌注后表现出不同程度的退行性变化、蛋白质表达丧失和/或ATP/CP合成紊乱。

结论

在亚低温下长期保存后,心循环死亡心脏的心脏组织结构/功能完整性得到有力维持,蛋白质表达和细胞能量代谢得以保存,这表明21℃是在索马溶液中对心循环死亡心脏进行长期保存的理想温度。

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