本文選題：生物人工肝　切入點(diǎn)：急性肝功能衰竭　出處：《南方醫(yī)科大學(xué)》2017年博士論文　論文類型：學(xué)位論文

【摘要】：第一章食蟹猴急性肝功能衰竭模型的建立及評(píng)價(jià)目的:穩(wěn)定的、重復(fù)性好的大動(dòng)物急性肝衰竭(ALF)模型是人工肝研究不可或缺的平臺(tái)。本章研究旨在建立一種藥物和外科食蟹猴ALF模型。方法:1、16只食蟹猴隨機(jī)分為4組(A,B,C,D),分別經(jīng)外周靜脈注射0.30,0.25,0.20 + 0.05(間隔 24 h),0.20 g/kg 的 D-氨基半乳糖(D-gal);給藥后觀察受試猴的臨床表現(xiàn),生存時(shí)間,生化、凝血、顱內(nèi)壓(ICP)和肝臟的病理變化;2、6只食蟹猴行袖套式插管門靜脈-右腎靜脈轉(zhuǎn)流聯(lián)合膽總管結(jié)扎、切斷術(shù)(PRRS+CBDLT),術(shù)后觀察動(dòng)物的臨床表現(xiàn),生化、凝血和肝臟的病理和組織學(xué)變化。結(jié)果:1、給藥后受試猴均出現(xiàn)不同程度的精神萎靡,進(jìn)食減少、黃疽等表現(xiàn),生存時(shí)間ABC三組分別為56±8.7h、95±5.5h和99±2.2h,D組除1只136h死亡外,其余猴全部存活。血清肝酶、TBiL、Cr、BUN、血Amm等明顯升高,ALB水平降低,PT明顯延長(zhǎng),ICP增高等,病理學(xué)鑒定有明顯肝細(xì)胞片狀壞死、伴出血。2、術(shù)后實(shí)驗(yàn)猴血清AST、ALT、CK、LDH、ALP、TBiL、DBiL等明顯升高,PT明顯延長(zhǎng),ALB水平降低,術(shù)后第10天肝標(biāo)本病理學(xué)和組織學(xué)檢測(cè)出明顯的肝細(xì)胞壞死、變性、凋亡等炎癥反應(yīng)。結(jié)論:成功構(gòu)建了 D-gal誘導(dǎo)的藥物性食蟹猴ALF模型和PRRS + CBDLT誘導(dǎo)的簡(jiǎn)化外科食蟹猴ALF模型。第二章基于微囊化豬肝細(xì)胞的生物人工肝安全性及有效性的評(píng)估目的:以具有自主知識(shí)產(chǎn)權(quán)的往返翻轉(zhuǎn)灌注型生物反應(yīng)器為核心,裝載微囊化的豬肝細(xì)胞,組建生物人工肝(BAL),通過(guò)對(duì)食蟹猴ALF模型的救治實(shí)驗(yàn)對(duì)其安全性和有效性進(jìn)行評(píng)價(jià),為其進(jìn)一步臨床試驗(yàn)奠定基礎(chǔ)。方法:采用D-gal分次誘導(dǎo)建立食蟹猴藥物性ALF模型,離體膠原酶兩步灌流法分離西藏小型豬原代肝細(xì)胞,制備海藻酸鈉-聚賴氨酸(APA)裹肝細(xì)胞微囊,裝入往返翻轉(zhuǎn)灌注型生物反應(yīng)器中,構(gòu)建BAL,用于ALF食蟹猴的治療。15只ALF食蟹猴隨機(jī)分為三組:(1)ALF組(空白對(duì)照):模型猴僅觀察不治療;(2)BAL組:模型猴接受裝載微囊化豬肝細(xì)胞的BAL治療;(3)Sham組(設(shè)備對(duì)照):模型猴接入BAL系統(tǒng)體外循環(huán)6小時(shí),反應(yīng)器內(nèi)裝載不含細(xì)胞的APA空微囊。比較三組實(shí)驗(yàn)猴治療前后的生存時(shí)間、生化指標(biāo)、PT及ICP等變化。結(jié)果:治療過(guò)程中實(shí)驗(yàn)猴均耐受良好。與兩個(gè)對(duì)照組相比,BAL組受試猴生存時(shí)間分別延長(zhǎng)14小時(shí)、16小時(shí)。BAL可以顯著降低受試猴血清肝酶、TBiL、血Amm等濃度和ICP,具有明顯的肝支持作用。結(jié)論:基于微囊化豬肝細(xì)胞的生物人工肝系統(tǒng)能夠改善ALF食蟹猴的血生化指標(biāo)和ICP,延長(zhǎng)生存時(shí)間。
[Abstract]:Chapter 1 Establishment and evaluation of acute hepatic failure model of crab eating monkey objective: stable, The animal model of acute liver failure (ARF) with good reproducibility is an indispensable platform for the study of artificial liver. This chapter aims to establish a drug and surgical ALF model of crab-eating monkey. The clinical manifestations of the tested monkeys were observed after intravenously injection of 0.30g / kg D-galactosamine (0.20g / kg D galactosamine at 24 h interval). Survival time, Biochemistry, Blood Coagulation, Intracranial pressure (ICP) and pathological changes of liver. Six crab-eating monkeys were treated with cuff intubation of portal vein and right renal vein bypass combined with common bile duct ligation. PRRS CBDLTT was performed after operation. The clinical manifestations and biochemistry of the animals were observed after operation. Results the pathological and histological changes of blood coagulation and liver were observed in all the monkeys treated with different degrees of mental retardation, decreased food intake and yellow gangrene. The survival time of ABC group was 56 鹵8.7 h, 95 鹵5.5 h and 99 鹵2.2 h respectively, except that 1 monkey died at 136h. All of the other monkeys survived. The serum TBiLX, Amm, etc., and the levels of serum Amm were significantly increased and PT was significantly prolonged. The histopathological identification showed that there was obvious hepatocyte necrosis. After operation, the levels of ALB in the serum of experimental monkeys were significantly increased, and the levels of ALB were significantly prolonged and decreased. On the 10th day after operation, liver necrosis and degeneration were observed in liver specimens. Conclusion: the ALF model induced by D-gal and the simplified ALF model induced by PRRS CBDLT were successfully constructed. Chapter 2 is based on the safety and safety of bioartificial liver of microencapsulated porcine hepatocytes. Effectiveness assessment purpose: to focus on round-trip reverse perfusion bioreactors with autonomous intellectual property rights, The microencapsulated porcine hepatocytes were loaded into the bioartificial liver BALA. The safety and efficacy of the bioartificial liver were evaluated by the treatment experiment of the ALF model of the crab monkey. Methods: the drug-induced ALF model of crab monkey was established by D-gal fractionation, and the primary liver cells of Tibetan miniature pig were isolated by two steps perfusion of collagenase in vitro. Sodium alginate-polylysine (APA) encapsulated hepatocyte microcapsules were prepared and loaded into a reverse perfusion bioreactor. BAL.15 ALF capped monkeys were randomly divided into three groups: control group (blank control group: model monkey only observed and not treated). Model monkey was treated with BAL loaded with microencapsulated porcine hepatocytes. The model monkey was connected to the BAL system for 6 hours after cardiopulmonary bypass. APA empty microcapsules without cells were loaded in the reactor. The survival time before and after treatment was compared in three groups of experimental monkeys. Results: the experimental monkeys were well tolerated in the course of treatment. Compared with the two control groups, the survival time was prolonged by 14 hours or 16 hours. Conclusion: the bioartificial liver system based on microencapsulated porcine hepatocytes can improve the blood biochemical index and the survival time of ALF monkey.
【學(xué)位授予單位】：南方醫(yī)科大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2017
【分類號(hào)】：R575.3

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本文編號(hào)：1570994