Significant Improvement of Survival by Intrasplenic Hepatocyte Transplantation in Totally Hepatectomized Rats
Significant Improvement of Survival by Intrasplenic Hepatocyte Transplantation in Totally Hepatectomized Rats
复制标题
脾内肝细胞移植显着改善全肝切除大鼠的存活率
作者：
Birgit A. P. M. Vogels;M. Maas;Anne Bosma;R. Chamuleau
本文观察了脾内肝细胞移植（HTX）对慢性肝萎缩大鼠急性肝功能衰竭的治疗作用。大鼠在第-14天接受门腔静脉分流术以诱导肝萎缩，并在第0天接受全肝切除术作为急性肝衰竭的开始。在第-7天、第-3天或第-1天进行脾内肝细胞或假移植（每组n = 4至6）。在肝切除术后的一段时间内，平均动脉血压保持在80 mm Hg以上，并预防了低血糖。肝性脑病的严重程度通过临床分级和EEG频谱分析来评估，同时测定血氨和血浆氨基酸浓度以及“生存”时间。处死后对脾脏和肺进行组织学检查。脾内肝细胞移植导致所有移植组中临床分级的显著改善（p < 0.05），而仅在第-1天移植的组中观察到EEG左侧指数的显著改善（p < 0.05）。与全肝切除术前1天肝细胞移植相比，全肝切除术前3天和7天肝细胞移植的大鼠与假移植对照组相比，“存活”时间显著增加3倍和2倍：第-1天HTX：7.5 ± 0.3 h vs. 5.9 ± 0.6 h 05），第-3天的HTX：19.7 ± 3.7 h对比6.5 ± 0.3 h（p < 0.05），以及第-7天的HTX：13.8 ± 3.2 h对比6.3 ± 0.3 h（p < 0.05）。此外，在第-3天和第-7天肝细胞移植的大鼠在全肝切除术后显示出显著较低的血氨浓度（p < 0.0001）。处死后脾脏的组织学检查显示红髓中有肝细胞簇。在脾脏中存在3天和7天的肝细胞显示胆汁蓄积和开始坏死的斑点。目前的数据表明，在门腔静脉分流大鼠完全肝衰竭的硬模型中，脾内肝细胞移植能够显著延长“存活”时间2至3倍。这一观察人类应用的相关性进行了讨论。
The effect of intrasplenic hepatocyte transplantation (HTX) was studied in an experimental model of acute liver failure in rats with chronic liver atrophy. Rats underwent a portacaval shunt operation on Day -14 to induce liver atrophy, and underwent total hepatectomy on Day 0 as a start of acute liver failure. Intrasplenic hepatocyte or sham transplantation was performed on Day -7, -3, or -1 (n = 4 to 6 per group). During the period following hepatectomy, mean arterial blood pressure was maintained above 80 mm Hg and hypoglycaemia was prevented. Severity of hepatic encephalopathy was assessed by clinical grading and EEG spectral analysis, together with determination of blood ammonia and plasma amino acid concentrations, and “survival” time. Histological examination of the spleen and lungs was performed after sacrifice. Intrasplenic hepatocyte transplantation resulted in a significant improvement in clinical grading in all transplanted groups (p < 0.05), whereas a significant improvement in EEG left index was seen only in the group with transplantation on Day -1 (p < 0.05). In contrast to hepatocyte transplantation 1 day before total hepatectomy, rats with hepatocyte transplantation 3 and 7 days before total hepatectomy showed a significant 3- and 2-fold increase in “survival” time compared to sham transplanted controls: HTX at Day -1: 7.5 ± 0.3 h vs. 5.9 ± 0.6 h (p > 0.05), HTX at Day -3:19.7 ± 3.7 h vs. 6.5 ± 0.3 h (p < 0.05), and HTX at Day -7: 13.8 ± 3.2 h vs. 6.3 ± 0.3 h (p < 0.05). Furthermore, rats with hepatocyte transplantation on Day -3 and -7 showed significantly lower blood ammonia concentrations after total hepatectomy (p < 0.0001). Histological examination of the spleens after sacrifice showed clusters of hepatocytes in the red pulp. Hepatocytes present in the spleen for 3 and 7 days showed bile accumulation and spots of beginning necrosis. The present data show that in a hard model of complete liver failure in portacaval shunted rats, intrasplenic hepatocyte transplantation is able to prolong “survival” time significantly 2- to 3-fold. The relevance of this observation for human application is discussed.