辅助性部分原位肝移植治疗急性肝功能衰竭的实验研究

发布时间：2018-02-26 15:30

本文关键词： 肝移植小鼠急性肝功能衰竭模型凋亡肝再生　出处：《华中科技大学》2014年博士论文　论文类型：学位论文

【摘要】：第一部分小鼠急性肝功能衰竭模型的建立目的建立一种相对简便、可重复的,具有一定治疗时间窗的小鼠急性肝功能衰竭模型,为研究辅助性部分肝移植治疗急性肝功能衰竭以及有关急性肝功能衰竭的病理生理学机制提供理论工具。方法使用异氟烷和氧气吸入全身麻醉,在第一肝门阻断的情况下使用丝线结扎切除小鼠约82%体积的肝脏以及胆囊,只留小鼠肝脏中叶的右侧部分,其中常温下第一阻断肝门时间为15和25分钟。结果当第一肝门阻断时间为25分钟时,急性肝功能衰竭的小鼠术后48小时的生存率为22.2%,术后7天的生存率仅为16.6%,术后小鼠肝脏组织HE染色显示形态紊乱,大量炎症细胞和巨噬细胞浸润,剩余肝细胞发生明显凋亡,术后肝细胞严重水肿坏死,肝小叶结构紊乱。术后6h、24h和36h三个时间点中血浆ALT、AST、TB和血氨浓度相比空白对照组明显升高。结论用小鼠第一次建立了一种联合肝切除和温缺血导致的、可重复的、操作简便、无生物危害性的小动物急性肝功能衰竭模型,同时此模型不仅可以应用于辅助性部分原位肝移植对急性肝功能衰竭的治疗研究,也可以用于急性肝功能衰竭本身的病理生理和分子机制的研究。第二部分辅助性部分原位肝移植治疗小鼠急性肝功能衰竭的实验研究目的利用小鼠急性肝功能衰竭模型和辅助性部分原位肝移植两个模型,通过实验观察辅助性部分原位肝移植对小鼠急性肝功能衰竭的治疗效果。方法C57小鼠左外叶肝脏作为肝移植供体,使用4℃肝素生理盐水灌注供体肝脏,顺利取下后,门静脉采用22G袖套套入并用11-0线捆绑缝合,供肝的胆管使用PE材质的空心管套入后固定以备胆肠引流,供肝放入4℃的HTK中保存。建立C57小鼠急性肝功能衰竭模型后作为受体,供肝门静脉套入受体门静脉,供肝流出道静脉与受体的肝上下腔静脉左侧用11-0prolene线行端侧吻合。移植肝体积约占受体肝的35%。术后观察7天生存率、肝功能指标、自身肝细胞凋亡、供肝和自身肝再生等情况。结果急性肝功能衰竭组小鼠术后48小时的总体生存率为13.6%(3/22),辅助性部分肝移植组的术后48小时生存率为81.8%(27/33),两者存在明显统计学差异。在术后的结果对比中,辅助性部分原位肝移植组血浆中的ALT、AST、TB和血氨浓度与急性肝功能衰竭组相比有明显的降低,辅助性部分原位肝移植组的自身肝细胞的凋亡有显著的降低,辅助性部分原位肝移植组的自身肝比急性肝功能衰竭组的自身肝细胞坏死和肝小叶结构紊乱程度明显减轻,而且辅助性部分原位肝移植组的自身肝在供肝的支持下,发生明显的肝细胞分裂和再生。结论辅助性部分原位肝移植作为治疗急性肝功能衰竭的一种有效手段,其机制是在一定体积的辅助肝脏的支持下,通过减少自身肝细胞的凋亡、促进自身肝再生,恢复肝脏功能等途径来提高生存率,同时供肝在移植后会迅速发生再生,也对肝功能的恢复起到重要作用。
[Abstract]:The first part of the model of acute liver failure in mice
Objective to establish a relatively simple and reproducible mouse model of acute liver failure with a time window. It will provide a theoretical tool for studying the pathophysiology mechanism of auxiliary partial liver transplantation for the treatment of acute liver failure and acute liver failure.
Methods isoflurane and oxygen inhaled general anesthesia. After the first hepatic portal occlusion, 82% volumes of liver and gallbladder were removed by using silk thread ligature. Only the right part of the middle lobe of mice was left. The first time of blocking the porta hepatis was 15 and 25 minutes at room temperature.
Results when the first hepatic portal occlusion time was 25 minutes, the mice with acute liver failure after 48 hour survival rate was 22.2% after 7 days, the survival rate is only 16.6%, after the operation of mouse liver tissue HE staining showed that the morphology of disorder, a lot of inflammatory cells and macrophages, the remaining liver cells apoptosis. Postoperative liver cell edema and necrosis of the hepatic lobules disorder. Postoperative 6h, plasma ALT, 24h and 36h three time points AST, TB control group was significantly increased compared with the blood ammonia concentration.
Conclusion the mouse first established a combined hepatectomy and warm ischemia induced, repeatable, simple operation, no acute liver failure animal model of biological harm, at the same time, this model not only can be used in auxiliary partial orthotopic liver transplantation on the treatment of acute exhaustive decline of liver function, can also be used for study on acute liver failure pathophysiology and molecular mechanism itself.
Experimental study on second parts of auxiliary partial orthotopic liver transplantation in the treatment of acute liver failure in mice
Objective To observe the effect of auxiliary partial orthotopic liver transplantation on acute liver failure in mice by using two models of acute liver failure and auxiliary partial orthotopic liver transplantation.
Methods C57 mice left lateral lobe of the liver as donor liver transplantation, using 4 C heparin saline infusion of donor liver, successfully removed, 22G cuff sleeved and 11-0 wire bundle suture by portal vein, hepatic bile duct using PE material of the hollow pipe is sleeved in fixed for biliary enteric drainage in liver preservation 4 C HTK. To establish acute hepatic failure model in C57 mice as receptor for portal vein in portal vein for the receptor, the receptor of hepatic outflow tract vein and suprahepatic inferior vena cava left with the 11-0prolene line for end to side anastomosis. The liver volume accounted for about 35%. receptor of liver were observed after surgery within 7 days rate, liver function index, the liver cell apoptosis, liver and its liver regeneration and so on.
The results of mice with acute liver failure after 48 hours, the overall survival rate was 13.6% (3/22), auxiliary partial liver transplantation group after 48 hours survival rate was 81.8% (27/33), there was statistical difference. In contrast to the postoperative results of auxiliary partial orthotopic liver transplantation group in plasma ALT, AST, TB were significantly lower compared with the blood ammonia concentration and acute liver failure group, apoptosis of auxiliary partial orthotopic liver transplantation group the liver cells were significantly reduced, auxiliary partial orthotopic liver transplantation group the liver significantly reduced than acute liver failure group the necrosis of liver cells and the degree of disorder of hepatic lobules, and auxiliary partial orthotopic liver transplantation group the liver in the donor liver support, liver cell division and regeneration significantly.
Conclusion the auxiliary partial orthotopic liver transplantation is an effective treatment for acute liver failure, the mechanism is in a certain volume of liver support, by reducing the apoptosis of hepatocytes and promote liver regeneration, liver function recovery and other ways to improve the survival rate, while liver regeneration occurs rapidly in transplantation after the recovery of liver function play an important role.

【学位授予单位】：华中科技大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R657.3

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