Fan Lin, Fu Zhen, Xiong Yan, Ye Shaojun, Wang Yanfeng, Peng Guizhu, Ye Qifa
Zhongnan Hospital of Wuhan University, Institute of Hepatobiliary Diseases of Wuhan University, Transplant Center of Wuhan University, Hubei Key Laboratory of Medical Technology on Transplantation, Wuhan, Hubei 430071, P.R. China.
The 3rd Xiangya Hospital of Central South University, Research Center of National Health Ministry on Transplantation Medicine Engineering and Technology, Changsha, Hunan 410013, P.R. China.
Exp Ther Med. 2021 Mar;21(3):256. doi: 10.3892/etm.2021.9687. Epub 2021 Jan 25.
Establishing a steatotic liver transplantation animal model can be a challenging process, which requires complex microsurgical technologies. The present study established a novel rat model of stable steatotic liver transplantation for marginal liver graft research, which notably minimized the number of animals used for the experiment. Briefly, male Sprague-Dawley rats (n=90) were fed with a high-fat diet (HFD; 60%, kJ) or standard chow diet (SCD) for 8 weeks. The liver enzymes and lipid levels were assessed every week, and the degree of steatosis was determined via hematoxylin and eosin and Oil Red O staining. The results demonstrated that there were no significant differences in alanine aminotransaminase and aspartate aminotransferase levels between the SCD and HFD groups (P>0.05), whereas the level of plasma triglyceride (TG) increased by 1.76-fold in the HFD group at week 2, and progressively decreased to baseline levels by week 8. Significantly higher levels of TG were observed in the HFD group compared with the SCD group at week 2 (P<0.05). In addition, the levels of plasma glucose (P<0.05), portal insulin (P<0.05) and content of liver lipid (P<0.01) increased in the HFD group compared with the SCD group. After 6 weeks, the liver steatosis was successfully formed and stable. Consequently, a rat liver developed hepatic macrovesicular steatosis >60%, which was subsequently used for transplantation after double-lobectomy. Post-transplantation survival rates in the HFD and SCD groups were as follows: Week 1, 80 vs. 100% and 1 month, 20 vs. 100%. A total of 20 rats were not sacrificed by performing double-lobectomy for biopsy. Taken together, the results of the present study suggest that rat liver double-lobectomy may be safely applied in steatotic liver transplantation without the need to sacrifice a large number of animals.
建立脂肪变性肝移植动物模型可能是一个具有挑战性的过程,这需要复杂的显微外科技术。本研究建立了一种用于边缘性肝移植研究的新型稳定脂肪变性肝移植大鼠模型,该模型显著减少了实验所用动物的数量。简要来说,将90只雄性Sprague-Dawley大鼠分为两组,一组喂食高脂饮食(HFD;60%,kJ),另一组喂食标准饲料(SCD),持续8周。每周评估肝酶和血脂水平,并通过苏木精-伊红染色和油红O染色确定脂肪变性程度。结果表明,SCD组和HFD组之间的丙氨酸氨基转移酶和天冬氨酸氨基转移酶水平无显著差异(P>0.05),而HFD组在第2周时血浆甘油三酯(TG)水平增加了1.76倍,并在第8周逐渐降至基线水平。与SCD组相比,HFD组在第2周时观察到显著更高的TG水平(P<0.05)。此外,与SCD组相比,HFD组的血浆葡萄糖水平(P<0.05)、门静脉胰岛素水平(P<0.05)和肝脂质含量(P<0.01)均有所增加。6周后,成功形成并稳定了肝脏脂肪变性。因此,一只大鼠肝脏出现了大于60%的肝大泡性脂肪变性,随后在双叶切除术后用于移植。HFD组和SCD组的移植后存活率如下:第1周,分别为80%和100%;1个月时,分别为20%和100%。共有20只大鼠未因进行双叶切除术用于活检而被处死。综上所述,本研究结果表明大鼠肝脏双叶切除术可安全应用于脂肪变性肝移植,而无需牺牲大量动物。