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中华肝脏外科手术学电子杂志 ›› 2023, Vol. 12 ›› Issue (01): 103 -107. doi: 10.3877/cma.j.issn.2095-3232.2023.01.020

基础研究

供肝免冲洗灌注的大鼠肝移植实验研究
卢强1, 杨丽斐2, 刘康3, 余佳薇2, 任璐4, 张谞丰3, 吕毅3,()   
  1. 1. 710061 西安交通大学第一附属医院老年外科;710061 西安交通大学第一附属医院精准外科与再生医学国家地方联合工程研究中心
    2. 710061 西安交通大学第一附属医院精准外科与再生医学国家地方联合工程研究中心
    3. 710061 西安交通大学第一附属医院精准外科与再生医学国家地方联合工程研究中心;710061 西安交通大学第一附属医院肝胆外科
    4. 710061 西安交通大学第一附属医院涉外病房
  • 收稿日期:2022-09-27 出版日期:2023-02-10
  • 通信作者: 吕毅
  • 基金资助:
    国家自然科学基金国家重大科学仪器研制项目(81727802); 陕西省重点研发计划高校联合项目-重点项目(2021GXLH-Z-047); 陕西省自然科学基础研究计划资助项目(2020JQ-528)

Experimental study of liver transplantation without graft irrigation or perfusion in rat models

Qiang Lu1, Lifei Yang2, Kang Liu3, Jiawei Yu2, Lu Ren4, Xufeng Zhang3, Yi Lyu3,()   

  1. 1. Department of Geriatrics, the First Affiliated Hospital of Xi'an Jiaotong University, Xi'an 710061, China; National and Local Joint Engineering Research Center of Precision Surgery and Regenerative Medicine, the First Affiliated Hospital of Xi'an Jiaotong University, Xi'an 710061, China
    2. National and Local Joint Engineering Research Center of Precision Surgery and Regenerative Medicine, the First Affiliated Hospital of Xi'an Jiaotong University, Xi'an 710061, China
    3. National and Local Joint Engineering Research Center of Precision Surgery and Regenerative Medicine, the First Affiliated Hospital of Xi'an Jiaotong University, Xi'an 710061, China; Department of Hepatobiliary Surgery, the First Affiliated Hospital of Xi'an Jiaotong University, Xi'an 710061, China
    4. Foreign Patient Ward, the First Affiliated Hospital of Xi'an Jiaotong University, Xi'an 710061, China
  • Received:2022-09-27 Published:2023-02-10
  • Corresponding author: Yi Lyu
目的

探讨供肝免冲洗灌注的大鼠肝移植的可行性。

方法

将72只健康成年SD大鼠随机分为3组:供体全身肝素化的供肝免冲洗灌注(NFLS组),供体不行全身肝素化的供肝免冲洗灌注(NFL组),供体行全身肝素化的供肝UW液冲洗灌注(对照组)。每组各24只,组内大鼠随机作为供受体进行同种异体大鼠肝移植。分别于移植术后24、72 h获取组织及血清标本送检,检测血清AST、ALT及TB;同时观察各组供肝术后大体观和组织病理学变化。3组AST、TB等肝功能指标比较采用Kruskal-Wallis秩和检验,两两比较采用Wilcoxon检验。

结果

3组均顺利完成肝移植,术后受体均顺利存活。NFLS组受体术后72 h血清AST、TB水平分别为112(89,181)U/L、0.5(0.4,0.6)μmol/L,明显低于对照组的453(361,692)U/L、1.8(1.5,2.2)μmol/L(Z=-2.56,-2.89;P<0.05)。3组术后24、72 h移植肝灌注良好,移植肝表面均未见明显坏死灶。HE染色显示NFLS组和对照组移植肝肝小叶结构正常,仅有轻微肝脏小叶中心微空泡脂肪变性;NFL组移植肝肝窦内部可观察到微循环血栓形成。

结论

与传统的器官保存液冲洗灌注相比,免冲洗灌注大鼠肝移植术后肝功能和组织学损伤并无明显加重,供肝免冲洗灌注肝移植可能具有一定可行性,供体全身肝素化是其关键条件。

Objective

To investigate the feasibility of liver transplantation without liver graft irrigation or perfusion in rat models.

Methods

72 healthy adult SD rats were randomly and evenly divided into three groups: non-flushed donor liver with systemic heparinization (NFLS group), non-flushed donor liver without systemic heparinization (NFL group), and systemic heparinization with UW solution irrigation and perfusion (control group). In each group, the rats were randomly assigned as liver transplantation donors and recipients. The tissue and serum samples were obtained at 24 h and 72 h after liver transplantation for AST, ALT and TB detection. Gross and histopathological changes of liver grafts were observed after liver transplantation. The liver function indexes, such as AST and TB, were compared by Kruskal-Wallis test, and pairwise comparison was performed by Wilcoxon test.

Results

Liver transplantation was successfully completed in all three groups, and all recipients survived after surgery. In the NFLS group, serum AST and TB levels were 112(89, 181) U/L and 0.5(0.4, 0.6) μmol/L at postoperative 72 h, significantly lower than 453(361, 692) U/L and 1.8(1.5, 2.2) μmol/L in the control group (Z=-2.56, -2.89; P<0.05). At postoperative 24 and 72 h, the liver grafts were properly perfused in three groups, and no evident necrosis was found on the surface of liver grafts. HE staining showed that the structure of liver lobules of grafts was normal in the NFLS and control groups, with slight fatty degeneration of microvesicles in the center of liver lobules. In the NFL group, microcirculatory thrombosis could be observed in the sinus of liver grafts.

Conclusions

Compared with conventional irrigation and perfusion using organ preservation solution, liver function and histological damage are not significantly aggravated after undergoing liver transplantation without irrigation or perfusion in rats, which might bea feasible option in clinical practice. Systemic heparinization of the donor is the key procedure.

表1 NFLS、NFL和对照组大鼠肝移植受体术后血清肝功能指标变化[MQ1Q3)]
图1 三组大鼠肝移植受体术后24、72 h肝脏大体观 注:3组移植肝术后24、72 h灌注良好,表面未见明显坏死灶;NFLS组为供体行全身肝素化供肝免灌注组,NFL组为供体不行全身肝素化供肝免灌注组,对照组为供体进行全身肝素化,供肝采用UW液进行冲洗灌注
图2 三组大鼠肝移植受体术后24、72 h肝脏组织学检查 注:HE染色显示NFLS组和对照组移植肝的肝小叶结构正常,仅有轻微肝小叶中心微空泡脂肪变性(<5%肝细胞);NFL组移植肝肝窦内可观察到微循环血栓形成;NFLS组为供体行全身肝素化供肝免灌注组,NFL组为供体不行全身肝素化供肝免灌注组,对照组为供体进行全身肝素化,供肝采用UW液进行冲洗灌注
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