低温机械灌注对大鼠心脏死亡器官捐献供肝移植的保护作用  

Protective effect of hypothermic machine perfusion in the rat from donor after cardiac death liver transplantation

作　　者：程楚红 范林[1] 付贞[1] 王彦峰[1] 熊艳[1] 叶殷发[1,2] 

机构地区：[1]武汉大学中南医院武汉大学肝胆疾病研究院武汉大学移植医学中心移植医学技术湖北省重点实验室,430071 [2]中南大学湘雅三医院卫生部移植医学工程技术研究中心,长沙410013

出　　处：《中华实验外科杂志》2017年第6期1008-1010,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金新疆联合基金（U1403223）;湖北省自然科学基金（2015CFA018）

摘　　要：目的建立稳定的大鼠心脏死亡器官捐献（DCD）供肝低温机械灌注后肝移植模型，探讨低温机械灌注（HMP）对大鼠DCD供肝移植的保护作用。方法雄性SD大鼠90只，分为3组，HMP组DCD30min后的肝脏用4℃HTK液冷保存（SOS）2h、低温机械灌注2h后行同种异体肝移植，SCS组DCD30min后的肝脏用4℃HTK液冷保存4h后行同种异体肝移植，正常对照组供肝不经过热缺血经灌洗后用4℃HTK液保存45min后行同种异体肝移植，观察低温机械灌注与冷保存对大鼠DCD供肝移植的影响。结果HMP组肝移植术后16h血清ALT含量为（2．04±0．41）IU／gliver，低于SCS组（4．51±1．07）IU／gliver，差异有统计学意义（P=0．001）；HMP组肝移植术后16h血清AST含量为（2．61±0．59）IU／gliver，低于SCS组（6．35±1．21）IU／gliver，差异有统计学意义（P=0．001）；HMP组低温机械灌注2h后Krtipple样转录因子2（KLF2）和内皮型一氧化氮合酶（eNOS）蛋白表达较灌注前明显增多，SCS组在后2h冷保存前后KLF2和eNOS蛋白表达无明显变化。结论低温机械灌注可使KLF2和eNOS蛋白表达增加，保护大鼠肝细胞，有利于提高供肝质量。Objective To develop a stable model of hypothermic machine perfusion （HMP） on liver grafts of rat from donor after cardiac death （DCD） in liver transplantation and study the protective effect of HMP during it. Methods Ninety male Sprague - Dawley （SD） rats （250 - 320 g） were random- ly divided into three groups ： hypothermic machine perfusion （HMP） group with 30 rain of in situ warm is- chemia ＋ 2 - h static cold storage followed by 2 - h hypothermic machine perfusion in HTK solution （4℃ ） （ n = 30）, static cold storage （SCS） group with 30 min in situ warm ischemia and 4 - h static cold storage in HTK solution （4℃ ） （ n = 30） , and normal control experiments with rat livers cold flushed and stored in HTK solution （4 ℃ ） immediately after procurement, i. e. , without in situ warm ischemia. Minimal static cold storage （45 min） was granted in this group to allow cuff placement before liver transplantation （ n = 30）. The effects of hypothermic machine perfusion and static cold storage were evaluated in the rat DCD liver transplantation. Results The ALT and AST levels in the plasma tested 16 hours after orthotopic liver transplantation in the HMP group were （2. 04 ±0. 41 ） IU/g liver, （2. 61 ±0. 59） IU/g liver respec- tively, while in the SCS group [ （4. 51 ± 1.07） IU/g liver, P = 0. 001 ], [ （6. 35 ± 1.21 ） IU/g liver, P = 0. 001 ] were increased as compared with HMP group. The expressions of krtipple like lactor 2 （KLF2） and endothelial nitric oxide synthase （eNOS） increased after HMP in the HMP group, while in the SCS group the expressions had no significant change during the last 2 - h static eold storage. Conclusion Hy- pothermic machine perfusion can increase the expressions of KLF2 and eNOS, and proteet rat hepatoeytes, and improve the quality of DCD liver.

关 键 词：大鼠 心脏死亡器官捐献供肝 低温机械灌注 冷保存 肝移植 

分 类 号：R657.3[医药卫生—外科学]