摘要
目的采用体外膜肺氧合(Extracorporeal membrane oxygenation,ECMO)全血离体灌注边缘性供肝,评估其改善肝脏功能的可行性。方法取6只比格犬按随机数字表法分为实验组和对照组,各3只。对照组犬肝脏常温下热缺血20 min后,用组氨酸-色氨酸-酮戊二酸盐(HTK)器官保存液灌注并冷保存2 h;实验组犬肝脏常温下热缺血20 min后,采用ECMO转流,常温下全血离体连续灌注2 h,用HTK器官保存液灌注并冷保存2 h。HE染色观察两组肝脏组织病理形态,透射电镜观察两组肝细胞及细胞器病理变化;免疫组化SABC法染色,组织化学评分法(Histochemistry score,H-Score)比较两组肝组织中肿瘤坏死因子(TNF-α)的表达水平。结果HE染色结果显示,犬肝热缺血20 min,两组肝组织内均出现大量肝细胞肿胀,肝窦内及汇管区可见大量出血;电镜下显示肝细胞核的核仁消失,线粒体数量减少,粗面内质网上附着的核糖体减少。对照组犬肝组织经HTK冷保存2 h后,肝小叶结构不完整,肝窦广泛扩张,大量肝细胞胞质内可见圆形空泡;电镜下可见肝细胞核异形,核仁消失,线粒体形状不规则,线粒体嵴略模糊部分嵴消失;粗面内质网有脱颗粒及灶性溶解发生。实验组犬肝组织经ECMO全血灌注2 h,HTK冷保存2 h后肝小叶结构大致完整,肝细胞肿胀较多,部分肝细胞有空泡状。电镜下肝细胞核圆而饱满,核仁明显,线粒体数量丰富、分布均匀且形状规则,粗面内质网未见明显脱颗粒。对照组在HTK冷保存2 h后,犬肝组织中TNF-α表达的H-Score评分显著高于正常肝组织和热缺血20 min后,差异有统计学意义(P<0.05);实验组在HTK冷保存2 h后,犬肝组织中TNF-α表达的H-Score评分显著低于对照组,差异有统计学意义(P<0.05)。结论ECMO全血离体灌注技术可有效减轻离体供肝的缺血再灌注损伤,有助于提高边缘性供肝的质量。
Objective To analyze the protective efficacy of extracorporeal membrane oxygenation(ECMO)with whole blood reperfusion in vitro for resuscitation of marginal donor liver after cardiac death(DCD)organ donation.Methods 6 beagles were divided into the experiment group and the control group,according to the principle of random envelope drawing,each group with 3 beagles.Both of groups were given total hepatectomy.After canine hepatic warm ischemia lasted for 20 minutes with room temperature,the livers in the control group were perfused once and preserved with cold histidine-tryptophan-ketoglutarate(HTK)solution with 0~4°C for 2 h.The livers in the experiment group were firstly underwent 2 h ECMO with 37°C whole-blood perfusion in vitro,and then it perfused and preserved with cold HTK solution same as the control group.Specimens of the liver tissue were taken,and the changes of liver structure were observed by hematoxylin-eosin(HE)staining;hepatocyte structure were observed by transmission electron micro⁃scopic(TEM)examination;expressions of TNF-αin liver tissue were detected by immunohistochemistry that were sub⁃sequently converted into continuous variable by histochemistry score(H-Score).Results After 20 min canine hepatic warm ischemia with room temperature,the most significant light microscopic histopathologic findings were extensive hepatocellular swelling,massive congestion in hepatic sinus and portal area of liver tissue,and TEM examination showed that hepatocyte nucleoli was disappeared,numerber of mitochondria and ribosomes attached to rough endoplas⁃mic reticulum with decreasing.After 2 h cold HTK solution preservation,light microscopic histopathologic changes were sensible damge of hepatic lobule,massive expansion of the hepatic sinus,extensive hepatocellular vacuolar de⁃generation.TEM examination showed that hepatic nuclear atypia with vanished nucleolus appeared,mitochondrion cristae fragmentation and part disappeared,degranulation and focal dissolution of rough endoplasmic reticulum al⁃ready o
作者
季学闻
马利兵
若巴提·艾尔肯
肉斯太木江·依马木
赵晋明
JI Xuewen;MA Libing;Ruobati Aierken;Rousitaimujiang Yimamu;ZHAO Jinming(Department of Liver and Laparoscopic Surgery,the First Affiliated Hospital of Xinjiang Medical University,Urumqi 830054,China)
出处
《新疆医科大学学报》
CAS
2022年第6期610-614,共5页
Journal of Xinjiang Medical University
基金
新疆维吾尔自治区自然科学基金(2016D01C273)。
关键词
体外膜肺氧合
缺血再灌注损伤
犬
肝组织
extracorporeal membrane oxygenation
ischemia reperfusion injury
dog
hepatic tissue
