低温机器灌注对肝脏移植物的保护涉及大鼠移植物再生的恢复。
Graft protection of the liver by hypothermic machine perfusion involves recovery of graft regeneration in rats.
作者信息
Jia Jun-Jun, Xie Hai-Yang, Li Jian-Hui, He Yong, Jiang Li, He Ning, Zhou Lin, Wang Weilin, Zheng Shu-Sen
机构信息
1 Department of Hepatobiliary and Pancreatic Surgery, First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, China.
*These authors contributed equally to this work.
出版信息
J Int Med Res. 2019 Jan;47(1):427-437. doi: 10.1177/0300060518787726.
OBJECTIVE
This study was performed to evaluate the impact and underlying mechanisms of hypothermic machine perfusion (HMP) on half-size liver graft regeneration.
METHODS
Forty rats were randomly assigned to five groups: two in vitro groups (static cold storage [SCS] and HMP) and three in vivo groups (orthotopic liver transplantation, SCS, and HMP). Perfusates and plasma samples were collected for analysis of hepatic enzymes. Liver tissue was obtained for evaluation of histology, immunohistochemistry (Ki67 and proliferating cell nuclear antigen [PCNA]), and the regeneration rate. Cell cycle genes were analyzed by quantitative real-time polymerase chain reaction, and cyclin D1 and cyclin E1 were semiquantified by western blot.
RESULTS
HMP improved histopathological outcomes and decreased hepatic enzyme release. The expression of Ki67 and PCNA demonstrated a greater proliferation activity in the HMP than SCS group, and the expression of almost all cell cycle genes was elevated following HMP. Western blot results showed higher protein levels of cyclin D1 and cyclin E1 in the HMP than SCS group.
CONCLUSIONS
Our findings suggest for the first time that half-size liver graft protection by HMP involves recovery of graft regeneration.
目的
本研究旨在评估低温机器灌注（HMP）对半肝移植肝再生的影响及潜在机制。
方法
40只大鼠随机分为五组：两个体外组（静态冷藏[SCS]和HMP）和三个体内组（原位肝移植、SCS和HMP）。收集灌注液和血浆样本以分析肝酶。获取肝组织以评估组织学、免疫组织化学（Ki67和增殖细胞核抗原[PCNA]）以及再生率。通过定量实时聚合酶链反应分析细胞周期基因，并通过蛋白质印迹法对细胞周期蛋白D1和细胞周期蛋白E1进行半定量分析。
结果
HMP改善了组织病理学结果并减少了肝酶释放。Ki67和PCNA的表达显示HMP组比SCS组具有更高的增殖活性，并且HMP后几乎所有细胞周期基因的表达均升高。蛋白质印迹结果显示HMP组中细胞周期蛋白D1和细胞周期蛋白E1的蛋白质水平高于SCS组。
结论
我们的研究结果首次表明，HMP对半肝移植的保护作用涉及移植肝再生的恢复。
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