發(fā)布時(shí)間：2018-02-26 15:30

  本文關(guān)鍵詞： 肝移植 小鼠 急性肝功能衰竭 模型 凋亡 肝再生　出處：《華中科技大學(xué)》2014年博士論文　論文類(lèi)型：學(xué)位論文

【摘要】：第一部分小鼠急性肝功能衰竭模型的建立 目的建立一種相對(duì)簡(jiǎn)便、可重復(fù)的,具有一定治療時(shí)間窗的小鼠急性肝功能衰竭模型,為研究輔助性部分肝移植治療急性肝功能衰竭以及有關(guān)急性肝功能衰竭的病理生理學(xué)機(jī)制提供理論工具。 方法使用異氟烷和氧氣吸入全身麻醉,在第一肝門(mén)阻斷的情況下使用絲線結(jié)扎切除小鼠約82%體積的肝臟以及膽囊,只留小鼠肝臟中葉的右側(cè)部分,其中常溫下第一阻斷肝門(mén)時(shí)間為15和25分鐘。 結(jié)果當(dāng)?shù)谝桓伍T(mén)阻斷時(shí)間為25分鐘時(shí),急性肝功能衰竭的小鼠術(shù)后48小時(shí)的生存率為22.2%,術(shù)后7天的生存率僅為16.6%,術(shù)后小鼠肝臟組織HE染色顯示形態(tài)紊亂,大量炎癥細(xì)胞和巨噬細(xì)胞浸潤(rùn),剩余肝細(xì)胞發(fā)生明顯凋亡,術(shù)后肝細(xì)胞嚴(yán)重水腫壞死,肝小葉結(jié)構(gòu)紊亂。術(shù)后6h、24h和36h三個(gè)時(shí)間點(diǎn)中血漿ALT、AST、TB和血氨濃度相比空白對(duì)照組明顯升高。 結(jié)論用小鼠第一次建立了一種聯(lián)合肝切除和溫缺血導(dǎo)致的、可重復(fù)的、操作簡(jiǎn)便、無(wú)生物危害性的小動(dòng)物急性肝功能衰竭模型,同時(shí)此模型不僅可以應(yīng)用于輔助性部分原位肝移植對(duì)急性肝功能衰竭的治療研究,也可以用于急性肝功能衰竭本身的病理生理和分子機(jī)制的研究。 第二部分輔助性部分原位肝移植治療小鼠急性肝功能衰竭的實(shí)驗(yàn)研究 目的利用小鼠急性肝功能衰竭模型和輔助性部分原位肝移植兩個(gè)模型,通過(guò)實(shí)驗(yàn)觀察輔助性部分原位肝移植對(duì)小鼠急性肝功能衰竭的治療效果。 方法C57小鼠左外葉肝臟作為肝移植供體,使用4℃肝素生理鹽水灌注供體肝臟,順利取下后,門(mén)靜脈采用22G袖套套入并用11-0線捆綁縫合,供肝的膽管使用PE材質(zhì)的空心管套入后固定以備膽腸引流,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為受體,供肝門(mén)靜脈套入受體門(mén)靜脈,供肝流出道靜脈與受體的肝上下腔靜脈左側(cè)用11-0prolene線行端側(cè)吻合。移植肝體積約占受體肝的35%。術(shù)后觀察7天生存率、肝功能指標(biāo)、自身肝細(xì)胞凋亡、供肝和自身肝再生等情況。 結(jié)果急性肝功能衰竭組小鼠術(shù)后48小時(shí)的總體生存率為13.6%(3/22),輔助性部分肝移植組的術(shù)后48小時(shí)生存率為81.8%(27/33),兩者存在明顯統(tǒng)計(jì)學(xué)差異。在術(shù)后的結(jié)果對(duì)比中,輔助性部分原位肝移植組血漿中的ALT、AST、TB和血氨濃度與急性肝功能衰竭組相比有明顯的降低,輔助性部分原位肝移植組的自身肝細(xì)胞的凋亡有顯著的降低,輔助性部分原位肝移植組的自身肝比急性肝功能衰竭組的自身肝細(xì)胞壞死和肝小葉結(jié)構(gòu)紊亂程度明顯減輕,而且輔助性部分原位肝移植組的自身肝在供肝的支持下,發(fā)生明顯的肝細(xì)胞分裂和再生。 結(jié)論輔助性部分原位肝移植作為治療急性肝功能衰竭的一種有效手段,其機(jī)制是在一定體積的輔助肝臟的支持下,通過(guò)減少自身肝細(xì)胞的凋亡、促進(jìn)自身肝再生,恢復(fù)肝臟功能等途徑來(lái)提高生存率,同時(shí)供肝在移植后會(huì)迅速發(fā)生再生,也對(duì)肝功能的恢復(fù)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.

【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2014
【分類(lèi)號(hào)】：R657.3

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本文編號(hào)：1538602