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THE MECHANISM OF LIVER FAILURE AFTER EXCESSIVE HEPATECTOMY

过度肝切除术后肝衰竭的机制

基本信息

批准号：
11671261
负责人：
TOGO Shinji
金额：
$ 1.41万
依托单位：
YOKOHAMA CITY UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2000
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11671261/
关键词：
apoptosis postoperative liver failure post operative liver failure

项目摘要

Massive hepatectomy often induced lethal hepatic failure. The mechanism has been described using two theories : indirect injury caused by microcircular disturbance induces necrosis to hepatocytes, and direct injury caused by cytotoxic disturbance induces apoptosis to hepatocyte.Exp.1We investigated the mechanism using our original experimental rat partial hepatectomy (PHx) models.Method : We use male Wistar rats. Rats were anesthetized with diethylether, performed 90%PHx and 95%PHx. The rats were divided into the following two groups : group 1, 90%PHx, as a maximum procedure of hepatectomy model, group 2, 95%PHx, as a lethal hepatic failure model. We investigated serum concentration of interleukin (IL)-1b, IL-6, tumor necrosing factor (TNF)-a, as a index of hypercytokinemia. Histological findings of remnant liver were also examined by hematoxylyn and eosin (HE) staining. Apoptotic hepatocytes were determined by using DAPI and TUNEL assay. Bcl-x protein (anti-apoptotic member of the Bcl … More -2 family) expression of remnant liver was studied by western blot analysis.Result : Serum concentrations of IL-1b, IL-6, TNF-a, were higher in group 2 than in group 1. HE staining showed that more degeneration hepatocytes and little mitosis appeared in group 2. Apoptotic hepatocytes were more in group 2 than in group 1. Bcl-x protein was expressed more in group 1 than in group 2.Conclusion : Apoptosis is one of the most important factors in the hepatic failure after excessive hepatectomy.Exp.2We applicated cDNA micoarray analysis in this models to clarify the mechanism of hepatic failure after excessive hepatectomy. The cell cycle of hepatocyte was stopped by overexpression of p21, ubiquitin and many cyclins. Apoptosis of hepatocyte was progressed by overexpression of Fas, many caspases and cytochrome C.Furthermore, genes of heat shock protein which protected from liver injury were rexpressed in 95%PHx group.Exp.3Results of Exp.1 demonstrate that expression of Bcl-xL protein as an anti-apoptotic factor or regeneration factor contributes to survival after 90%PHx. We therefore transfected human bcl-2 gene (hbcl-2) to DA rat livers by an adenovirus vector. The hbcl-2 was efficiently expressed. 95%PHx was then performed. Liver damage was improved and the apoptotic cell count decreased, but the rats died.We concluded that transfection of hbc1-2 gene partly prevents cytotoxity (apoptosis), but cannot ensure survival. Thus, some other factor is required (e.g, a regeneration stimulator) to maintain life in these models. Less

大面积肝切除术常引起致死性肝衰竭。其机制有两种说法：微环干扰引起的间接损伤引起肝细胞坏死，细胞毒性干扰引起的直接损伤引起肝细胞凋亡。我们使用原始的实验性大鼠部分肝切除术（PHx）模型来研究其机制。方法：选用雄性Wistar大鼠。用二乙醚麻醉大鼠，分别给予90%PHx和95%PHx。将大鼠分为两组：1组，90%PHx，作为肝切除术最大程序模型；2组，95%PHx，作为致死性肝衰竭模型。我们研究了白细胞介素(IL)-1b、IL-6、肿瘤坏死因子(TNF)-a的血清浓度，作为高细胞素血症的指标。并用苏木精和伊红（HE）染色检查残肝的组织学表现。采用DAPI法和TUNEL法检测肝细胞凋亡。western blot检测残肝Bcl-x蛋白（Bcl…More -2家族抗凋亡成员）的表达。结果：2组大鼠血清IL-1b、IL-6、TNF-a浓度均高于1组。HE染色显示2组变性肝细胞增多，有丝分裂少。2组肝细胞凋亡明显多于1组。Bcl-x蛋白在1组的表达高于2组。结论：细胞凋亡是过度肝切除术后肝衰竭的重要因素之一。我们在这些模型中应用cDNA微阵列分析来阐明过度肝切除术后肝衰竭的机制。p21、泛素及多种细胞周期蛋白的过表达使肝细胞周期停止。过量表达Fas、多种半胱天冬酶和细胞色素c可促进肝细胞凋亡，95%PHx组可表达保护肝损伤的热休克蛋白基因。3 Exp.1的结果表明，Bcl-xL蛋白作为抗凋亡因子或再生因子的表达有助于90%PHx后的存活。因此，我们通过腺病毒载体将人bcl-2基因（hbcl-2）转染到DA大鼠肝脏。hbcl-2高效表达。95%PHx。大鼠肝损伤改善，凋亡细胞计数减少，但死亡。我们得出结论，转染hbc1-2基因部分阻止细胞毒性（细胞凋亡），但不能确保存活。因此，需要一些其他因素（例如，再生刺激器）来维持这些模型中的生命。少