Department of Surgery, Keio University, Tokyo, Japan.
Department of Pathology, Keio University, Tokyo, Japan.
Sci Rep. 2018 Oct 9;8(1):14987. doi: 10.1038/s41598-018-32889-y.
A feasible large animal model to evaluate regenerative medicine techniques is vital for developing clinical applications. One such appropriate model could be to use retrorsine (RS) together with partial hepatectomy (PH). Here, we have developed the first porcine model using RS and PH. RS or saline control was administered intraperitoneally to Göttingen miniature pigs twice, two weeks apart. Four weeks after the second dose, animals underwent PH. Initially, we tested different doses of RS and resection of different amounts of liver, and selected 50 mg/kg RS with 60% hepatectomy as our model for further testing. Treated animals were sacrificed 3, 10, 17 or 28 days after PH. Blood samples and resected liver were collected. Serum and liver RS content was determined by Liquid Chromatograph-tandem Mass Spectrometer. Blood analyses demonstrated liver dysfunction after PH. Liver regeneration was significantly inhibited 10 and 17 days after PH in RS-treated animals, to the extent of 20%. Histological examination indicated hepatic injury and regenerative responses after PH. Immunohistochemical staining demonstrated accumulation of Cyclin D1 and suppression of Ki-67 and PCNA in RS-treated animals. We report the development of the first large animal model of sustained liver injury with suppression of hepatic regeneration.
评估再生医学技术的可行大型动物模型对于开发临床应用至关重要。一种合适的模型可以是使用反式丝裂霉素（RS）与部分肝切除术（PH）相结合。在这里，我们使用 RS 和 PH 开发了第一个猪模型。RS 或生理盐水对照通过腹腔内给药两次，间隔两周。第二次剂量后 4 周，动物接受 PH。最初，我们测试了不同剂量的 RS 和不同数量的肝切除术，并选择 50mg/kg RS 进行 60%肝切除术作为我们的模型进行进一步测试。在 PH 后 3、10、17 或 28 天处死处理后的动物。采集血样和切除的肝脏。通过液相色谱-串联质谱法测定血清和肝脏 RS 含量。血液分析表明 PH 后肝功能障碍。PH 后 10 天和 17 天，RS 处理的动物的肝再生明显受到抑制，达到 20%。组织学检查表明 PH 后肝损伤和再生反应。免疫组织化学染色表明 RS 处理的动物中 Cyclin D1 积累和 Ki-67 和 PCNA 抑制。我们报告了第一个具有抑制肝再生的持续肝损伤的大型动物模型的开发。
