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对马斯特里赫特Ⅱ类供猪应用体外膜肺氧合修复供肝最佳时间的研究

Optimal repairing time of extracorporeal membrane oxygenation to Maastricht Ⅱ swine liver

摘要:

目的 研究对热缺血30 min的马斯特里赫特Ⅱ类(M-Ⅱ类)供猪应用体外膜肺氧合(ECMO)修复供肝的最佳时间.方法 巴马小型猪36只,采用随机数字表法分为3组,ECMO修复2h组、4h组和6h组,每组12只,供、受猪各6只.供猪以静脉推注氯化钾的方法诱导心跳骤停,给予标准心肺复苏30 min后宣布死亡.经下腔静脉及腹主动脉插管连接ECMO进行自体血循环.2h组、4h组和6h组分别运行ECMO2 h、4h和6h.监测记录肝功能、门静脉和肝动脉血流、动脉血气等指标.取肝组织甲醛固定后切片,HE染色,在光学显微镜下观察病理改变,采用脱氧核糖核苷酸末端转移酶介导的缺口末端标记法(TUNEL)检测肝细胞凋亡,并计算凋亡指数.ECMO运行结束后获取供肝,于UW液中冷保存2h,行原位肝移植手术.术后1~5 d取受猪外周血检测天冬氨酸转氨酶(AST)和丙氨酸转氨酶(ALT)水平.结果 随着ECMO运行时间的增加,肝功能指标和乳酸水平持续降低.ECMO运行结束时,6h组的门静脉血流量低于其他两组,差异有统计学意义(P<0.05).2h组、4h组和6h的凋亡指数分别为(40.20±7.22)%、(18.60±4.04)%和(29.25±5.98)%,与其他两组比较,4h组凋亡指数显著降低(P<0.05),且4h组肝组织的病理改变也明显较轻.术后5d,2h组和6h组受猪存活率均为83%,4h组为100%,术后4h组的转氨酶水平低于2h组和6h组,差异有统计学意义(P<0.05).结论 ECMO修复M-Ⅱ类供肝4h时修复效果较好,修复时间过短修复效果不理想,超出4h后肝脏功能与肝细胞活力出现下降.

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abstracts:

Objective To investigate the optimal time of extracorporeal membrane oxygenation (ECMO) on repairing Maastricht Ⅱ pig liver with warm ischemia injury for 30 min.Method Thirty-six miniature pigs were randomized to ECMO 2-h group,ECMO 4-h group and ECMO 6-h group,12 pigs per group,6 donors and 6 recipients.Cardiac arrest was induced by administration of 1 g KCl intravenously,followed by cardiopulmonary resuscitation (CPR) for 30 min according to the standard guideline.Cannulas was placed through inferior vena cava and abdominal aorta,then connected to ECMO extracorporeal circulation pipes.Transaminase,circulation flow rate of portal vein and hepatic artery and arterial blood gas were monitored and recorded.The hepatic tissues were cut into sections for pathological observation by HE stain under a light microscope.Apoptosis was detected by TUNEL and apoptotic index was calculated.The liver was stored in cold UW for 2 h after the ECMO circulation,then orthotopic liver transplantation without veno-venous bypass was performed.The levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were measured in the peripheral blood for 5 days after the operation.Result With the increase of the running time of ECMO,transaminase and lactate levels were decreased continuously.Circulation flow rate of portal vein and hepatic artery in ECMO 6-h group was significantly lower than that in the other two groups (P<0.05).Pathological change in ECMO 4-h group was milder than the rest two groups.Apoptosis index in ECMO 2-h,4-h and 6-h groups was (40.20 ± 7.22)%,(18.60 ± 4.04)% and (29.25 ± 5.98) %,respectively.The 5-day suvival rate in ECMO 2-h,4-h and 6-h groups was 83%,100% and 83%,respectively.The transaminase level in ECMO 4-h group at 5th day after the operation was lower than in ECMO 2-h group and 6-h group (P<0.05).Conclusion The optimal time of ECMO on repairing Maastricht Ⅱ liver was 4 h.The effect of restoration is not ideal when circulation time is not enough.Liver function and liver cell viability decline beyond 4 h.

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作者: 隋明昊 [1] 刘蕾 [2] 史源 [2] 马宁 [2] 刘艳 [2] 沈中阳 [2]
栏目名称: 实验研究
DOI: 10.3760/cma.j.issn.0254-1785.2015.05.008
发布时间: 2015-09-11
基金项目:
国家高技术研究发展计划(863)项目 天津市科技计划项目 天津市卫生行业重点攻关项目
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