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Objective We subject to establish a stable and practical normothermic machine perfusion （NMP）system, as well as to investigate the effect and its mechanism of NMP in alleviating the ischemia reperfusion injury of donation after cardiac death（DCD）grafts. Methods Ten Bama miniature pigs clamped right renal artery underwent 30 mins warm ischemia and were randomly allocated to two groups. In the group A, kidneys underwent 8-hour NMP preservation. In the group B, kidneys were immediately perfused with UW solution and preserved with this cold storage for 8 hours. kidney transplantation was performed in both groups. Then the two groups underwent autologous renal transplantation. The survival rate, the serum biochemical markers, the histopathological changes and the expression of cytokines were compared between these two groups. Results The level of tumor necrosis factor-α（TNF-α）, cysteinyl aspartate specific proteinase-3（Caspas-3）, endotoxin-1（ET-1）in group A was lower than those of group B, and the proportion of apoptotic cells measured by TUNEL method was also significantly lower than that of group B（P ＜ 0.05）, The levels of serum creatinine（SCr）, TNF-α and Caspase-3 in group A were significantly lower than those in group B when 6 hours after bloodstream was opened and the pathological examination demonstrated that the injury of the kidney in group A was slighter than that in group B after 8-hour preservation and reperfusion Conclusion NMP storage could significantly alleviate the ischemia reperfusion injury of pig DCD kidneys. Its mechanism was that NMP could mimic the physiological environment of organs, maintain the cellular metabolism, reduce the expression of inflammatory cytokines after preservation and reperfusion.

摘要：

目的 构建新型常温机械灌注（normothermic machine perfusion，NMP）系统，利用猪自体肾 移植模型，探讨 NMP 改善猪心脏死亡器官捐献（donation after cardiac death，DCD）供肾保存质量及相关机制。 方法 选用健康巴马小型猪 10 只，随机分为两组，在原位夹闭右肾动脉 30 分钟后，UW 液低温灌注，A 组 （n ＝ 5）以 NMP 保存 8 小时，B 组（n ＝ 5）以 UW 液 4℃低温保存 8 小时，然后两组均行自体肾移植术， 比较术后血清生化指标、肾组织病理变化、细胞因子的表达情况等指标。结果 两组均成功在经过 30 分钟 热缺血和 8 小时保存后，进行自体肾移植。血流开放 1 小时后，A 组肾组织免疫组化肿瘤坏死因子 -α（tumor necrosis factor-α，TNF-α）、天冬氨酸特异性半胱氨酸蛋白酶 -3（cysteinyl aspartate specific proteinase-3， Caspase-3）、内毒素（endotoxin-1，ET-1）水平明显低于 B 组，TUNEL 法测定凋亡细胞比例也明显低于 B 组， 两组之间差异有统计学意义（P ＜ 0.05）；血流开放 6 小时后，A 组血清肌酐（serum creatinine，SCr）、 TNF-α、Caspase-3 水平明显低于 B 组，两组之间差异有统计学意义（P ＜ 0.05）；开放后肾脏活检常规病 理检查提示，A 组肾脏损伤程度明显减轻。结论 NMP 保存可明显减轻猪 DCD 供肾的缺血 / 再灌注损伤 ； 其保护机制与 NMP 可模拟器官的生理环境，维持充足能量与氧气供应，减少保存期间和再灌注后炎性细胞 因子释放等有关。