國立臺灣大學醫學院外科袁瑞晃2006-07-262018-07-112006-07-262018-07-112001-07-31http://ntur.lib.ntu.edu.tw//handle/246246/24445肝臟是人體中極為重要的器官，但是 肝臟的疾病如B 型肝炎、C 型肝炎、肝硬 化、肝癌等向來是東方人尤其是中國人健 康極大的問題，肝臟疾病末期病人往往令 人束手無策，近年來雖然可以進行肝臟移 植，但由於捐贈者不多因此受益者有限， 因此希望可發展出新的技術以照顧肝臟疾 病末期病患。 在許多的研究發現肝臟有很強的再生 作用，切除百分之七十左右的肝臟也可以 再生，以往分離出肝細胞雖然可以加以培 養，但數代之後肝細胞便呈現老化現象無 法長期培養，因此尋找肝臟幹細胞也就是 所謂的卵圓細胞便顯的非常重要。由於目 前實驗所知對於肝臟的再生作用主要都是 原本存在的成熟肝細胞分裂所產生，而 Acetylaminofluorene (AAF)可以在肝臟中 被肝細胞代謝成有毒性的N-hydroxy 衍生 物，抑制成熟肝細胞的生長分裂，因而本 實驗希望探討除了阻斷大部分肝臟血流造 成肝細胞大量喪失後（阻斷肝臟血流的方 式是保留供給右葉之肝動脈及門靜脈，而 將肝臟中葉以及左葉的血流完全阻斷，由 先驅研究知道其相當於百分之七十左右的 肝臟切除），再藉由此藥物抑制肝成熟臟 細胞的再生作用，進而刺激肝臟卵圓細胞 的大量活化。 本實驗將小鼠任意分成四組，第一組 (V-group)不給AAF 而只經由手術綁掉血 管；第二組(AV-L-group)術前五天開始餵低 劑量AAF (5mg/kg)，綁完血管後繼續餵食 AAF 直到犧牲；第三組(AV-H-group)術前 五天開始餵高劑量AAF (7.5mg/kg)，綁完 血管後繼續餵食AAF 直到犧牲；第四組 (C-group)為控制組，只進行Sham operation，不餵食AAF 也不綁掉血管。各 組小鼠在分別術後第三、五、七、九及十 四天時犧牲，取其肝臟作分析。 至於肝臟細胞再生能力的分析是利用 BrdU incorporation 方式，而cytokeratin18, cytokeratin19, cytokeratin7, Thy-1 的測定則 用以分析肝臟卵圓細胞的活化現象。Liver is a very important visceral organ for the life. In the oriental country, especially in China and Taiwan, hepatic disease including hepatitis and liver cirrhosis is common and plays an important role in the health problem. For patients with end-staged liver disease, nothing can be done except liver transplantation. It’s not easy to have a Zeon PDF Driver Trialwww.zeon.com.tw 3 donor suitable for the recipient from time to time and further new ways for this problem is required in the near future. The liver is usually able to mount a prompt proliferative response to parenchymal loss and other hyperplastic stimuli. Following 70% partial hepatectomy in the rat, hepatocyte proliferation is greatly accelerated for a relatively short period while the deficit in hepatocyte number is replaced. Even though primary culture of the hepatocytes had been done for years the proliferative ability and viability decreased after several generations. So further researches for the hepatic stem cells that can be cultured for a long period and have the ability to differentiate to mature hepatocytes become more and more important. The principle in animal model is that when majority of the hepatic parenchyma is damaged, the ability of surviving hepatocytes can be regenerated quickly without activation of hepatic stem cells (oval cells). Further hepatic insult that inhibits hepatocyte regeneration is required for activation of oval cells. Acetylaminofluorene (AAF), an aromatic amine, can be metabolized in hepatocytes and resulted in N-hydroxy derivatives which was cytotoxic and mitoinhibitory for hepatocytes but not biliary cells or oval cells. The purpose of this study is to evaluate the additional effect of major hepatic vascular ligation (ligation of the vascular inflow to the middle and left lobes and preserve the right lobe only, that equal to about 70% partial hepatectomy in our preliminary study) and hepatotoxic agent administration in activation of hepatic oval cells. Animals were randomly assigned to four different groups. Mice receiving vascular occlusion only (V-group); mice receiving low dose (5mg/kg) of AAF and vascular ligation (AV-L-group); mice receiving high dose (7.5mg/kg) of AAF and vascular ligation (AV-H-group); and control group (C-group). AAF was given continuously until the mouse was sacrificed in all groups. After various time intervals (3, 5, 7, 9, 14 days) postoperatively, the mice were sacrificed and the liver was taken for further studies. BrdU incorporation with BrdU-FLUOS kit was used for regeneration evaluation and immunohistochemical studies about the cytokeratin18, cytokeratin19, cytokeratin7, Thy-1, were used to evaluate the activation of hepatic oval cell.application/pdf44603 bytesapplication/pdfzh-TW國立臺灣大學醫學院外科肝臟血流阻斷肝臟卵圓細胞cytokeratinThy-1hepatic vascular inflow ligationhepatic oval cell[SDGs]SDG3reporthttp://ntur.lib.ntu.edu.tw/bitstream/246246/24445/1/892314B002436.pdf