小型猪非转流下小体积移植肝损伤模型的建立及评价

Establishment and evaluation of models for small-for-size graft injury without veno-venous bypass in miniature pigs

目的建立标准化的、可行性强且临床贴合性好的小型猪小体积移植肝损伤模型，为小肝综合征的深人研究奠定基础。方法选用巴马小型猪，采用供体体内原位减体积70％的方法（切除肝左外叶、左中叶及大部右中叶），将30％残肝在非静脉转流下行同种异体原位肝移植术。记录动物手术相关数据、手术存活率及小肝7d存活率，计算术后7d移植肝再生百分比；监测术中血流动力学及血气改变；采集再灌注后2h、术后1～7d的外周血，检测血清总胆红素（TB）、谷丙转氨酶（ALT）、谷草转氨酶（AST）、凝血酶原时间及肌酐（Cr）水平的变化。猪死后当天尸检明确死亡原因。结果实验共完成12对次小型猪小体积肝移植术。所取供肝与受体全肝重量比（GW／RLW）为（28．63±4．42）％，与受体体重比（GW／BW）为（0．73±0．06）％。平均手术时间、无肝期及肝下下腔静脉阻断时间分别为（191．7±14．2）min、（28．3±3．6）min和（45．0±5．8）min。实验动物手术存活率83．33％（10／12），小肝7d存活率40％（4／10），7d移植肝再生百分比为（278．06±42．95）％。与无肝前期相比，无肝期动物心率明显加快，平均动脉压、中心静脉压、肛温（RT）、pH、BE值均显著下降，而血清K^＋。显著升高（P〈0．01），至关腹前RT、平均动脉压、中心静脉压、K^＋已基本恢复。血清ALK、AST、PT和Cr均于术后1d达到峰值，TB于术后第2天达到峰值，与灌注前水平比较具有显著性差异（P〈0．01）。至术后7d，除ALT、Cr基本恢复外，TBIL、AST和PT仍维持在较高水平。结论用巴马小型猪建立的非转流下30％体积供肝移植模型，具有标准化程度高、可行性强且临床贴合性好的优点，可作为小体积肝移植物损伤研究的理想大动物模型。

Objective To establish small-for-size （SFS） graft injury models in miniature pigs with high standardization, reproducibility and similarity to clinical situation. Methods Ba Ma minia- ture pigs were introduced in this study and orthotopic liver transplantations （OLTs） were performed in 12 pigs with 30% liver volume allogeneil grafts （small portion of right paramedian lobe, right lateral lobe and caudate lobe） without veno-venous bypass. The profiles of intra-operational hemodynamics and metabolism were investigated. Animals were observed for 7 days with daily serum biochemistry and coagulation function exam. The survival rate related to operation itself and the SFS grafts were respectively calculated as well as the graft regenerative ratio at post-operational day （POD） 7. Results Graft weight as a percentage of the recipient＇ s native liver weight （GW/RLW） and the total body weight （GW/BW） were （28.63±4.42） % and （0.73±0.06） %. The mean operation time, anhepatic phase, and the time of blockage of infra-hepatic IVC were （191.7±14.2） min, （28.3±3.6）min, and （45.0±5.8） min. The survival rate related to the operation itself and the SFS graft were 83. 33% （10/12） and 40% （4/10）, and the graft regenerative ratio at POD7 was （278.06±42.95）%. Contrast to the remarkable increase of heart rate and serum potassium during anbepatic phase, the mean arterial pressure, central venous pressure, rectal temperature, PH value and buffer excess had a significant decrease （P〈0.01） with a gradual recovery after reperfusion. Serum ALT, AST, PT, Cr, and TB were significantly increased with a peak level at POD1 for the former 4 and POD2 for TB, and then began to decrease and favorably recovered at POD7, but TB, PT, and AST levels were still high when compared to those of prereperfusion （P〈0.05）. Conclusion This model of OLT performed with 30% liver volume graft without veno-venous bypass was an ideal large animal model for series studies related to SFS graft injury.