THE MECHANISM OF LIVER FAILURE AFTER EXCESSIVE HEPATECTOMY INVESTIGATED USING CDAN MICROARRAY

使用 CDAN 微阵列研究过度肝切除术后肝衰竭的机制

• 批准号: 13671322
• 负责人: TOGO Shinji
• 金额: $ 1.66万
• 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: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13671322/
• 关键词: apoptosis; postoperative liver failure; cDNA Microarray; Liver regeneration; hepatectomy; cDNA microarra

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.1Excessive hepatectomy often causes fatal liver failure. We have reported that this is mainly mediated by apoptosis, characterized pathologically by TUNEL assay positive hepatocytes and a ladder pattern in DNA fragmentation assays. To investigate the mechanism, we used a cDNA microarray analysis to compare clearly differentiated rat partial hepatectomy (PHx) models (90%PHx, and 95%PHx). All 90%PHx rats survived, but the 95%PHx animals died of hepatic failure within 96 hours. The remnant liver was obtained at 4 time points (1,3,12, and 24 hrs after PHx). After RNA extraction, two samples were labeled with different fluorescent dyes and hybridized to the RIKEN set of 18,816 full-length enriched mouse cDNA arra … More ys. Scanning for fluorescent dye signals was performed, and many caspases were upregulated at 1 hr after PHx in the 95%PHx group. On the other hand, genes of Bcl-2, heat shock proteins and gluthatione-S-transferase were downregulated. We concluded that fatal hepatic after excessive hepatectomy was characterized by increased apoptosis and diminished liver regeneration.Exp.2Background/Aim : The liver has the capacity to regenerate after partial hepatectomy. In order to clarify the mechanism of liver regeneration, we observed the initial stage, especially the mechanism of gene expression during progress from G0 to S phase (0〜24 hrs), and attempted to identify the new gene controlling progress to the S phase.Methods : We applied large-scale gene expression analysis with complementary DNA (cDNA) microarrays in mouse hepatectomy models to clarify the mechanism of liver regeneration after partial hepatectomy.Results : As a result, 23 new immediate-early gene candidates such as IRAK-1 (interleukin-1 receptor associated kinase-1) and karyopherin alpha 1, which are involved in transportation within the nucleus, were discovered. Candidates for new genes concerned with the progress to the S phase were discovered : ID2 (inhibitor of DNA binding 2) and ID3 (inhibitor of DNA binding 3), both new liver regeneration factors that promoted progress to the S phase, and Gadd45 gamma (growth arrest and DNA-damage-inducible protein) as a factor inhibiting that process.Conclusions : The above result not only suggests the importance of NF-κB in the initial stage of liver regeneration but also points to the orderly maintenance of the proliferation of the cells in liver regeneration. Less

大面积肝切除术常引起致死性肝功能衰竭。其机制主要有两种学说：微循环紊乱引起的间接损伤导致肝细胞坏死，细胞毒性紊乱引起的直接损伤导致肝细胞凋亡。实验1过度肝切除常导致致死性肝功能衰竭。我们已经报道了这主要是由凋亡介导的，其病理特征是TUNEL检测阳性肝细胞和DNA片段化检测中的梯形图。为了研究这一机制，我们使用cDNA微阵列分析来比较明显分化的大鼠部分肝切除（PHx）模型（90%PHx和95%PHx）。所有90%PHx大鼠均存活，但95%PHx动物在96小时内死于肝功能衰竭。在PHx后1、3、12和24 h 4个时间点取剩余肝脏。RNA提取后，两个样品用不同的荧光染料标记，并与RIKEN的18，816个全长富集的小鼠cDNA阵列杂交。 ...更多信息 是的。扫描荧光染料信号，在95%PHx组中PHx后1小时，许多半胱天冬酶上调。Bcl-2、热休克蛋白和谷胱甘肽-S-转移酶基因表达下调。结论：过度肝切除后致死性肝损伤的特征是细胞凋亡增加和肝再生减少。实验2背景/目的：肝部分切除后肝脏具有再生能力。为了阐明肝再生的机制，我们观察了肝再生的起始阶段，特别是从G 0期到S期的基因表达机制（0 ~ 24小时），并试图鉴定控制向S期进展的新基因。我们应用互补DNA（cDNA）进行大规模基因表达分析，微阵列在小鼠肝切除模型中的应用，以阐明部分肝切除后肝再生的机制。结果，发现了23个新的立即早期基因候选者，如IRAK-1（白细胞介素-1受体相关激酶-1）和karyopherin alpha 1，它们参与细胞核内的运输。发现了与S期进展有关的新基因的候选者：ID 2（DNA结合抑制剂2）和ID 3（DNA结合抑制剂3），两种新的肝再生因子，促进进展到S期，和Gadd 45 γ（生长停滞和DNA损伤诱导蛋白）作为抑制该过程的因子。上述结果不仅提示NF-κB在肝再生初期的重要性，而且提示在肝再生过程中细胞增殖的有序维持。少

项目成果

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Fujii Y 等人：“门静脉栓塞术后肝切除术后肝功能衰竭的危险因素。”J Hep Bil Pancr Surg.. 10. 226-232 (2003)

Nishizuka I, Togo S et al.: "Analysis of Gene Expression Involved in Brain Metastasis from Brest Cancer Using cDNA Microarray"Brest Cancer. 9(1). 26-32 (2002)

Nishizuka I、Togo S 等人：“使用 cDNA 微阵列分析参与乳腺癌脑转移的基因表达”乳腺癌。

Fujii Y, Togo S et al.: "Risk factor of posthepatectomy liver failure after portal vein embolization"J Hepatobiliary Pancreat Surg. 10. 226-232 (2003)

Fujii Y、Togo S 等：“门静脉栓塞术后肝切除术后肝功能衰竭的危险因素”J Hepatobiliary Pancreat Surg。

Nagano Y, Togo S et al.: "Improved functional reserve of hypertrophied contra lateral liver after Portal vein ligation in rats"J.Hepatology. 37(7). 72-77 (2002)

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Shimizu D, Togo S et al.: "Current progress in the prediction of chemosensitivity for breast cancer"Breast Cancer. 11(1). 42-48 (2004)

Shimizu D、Togo S 等人：“乳腺癌化疗敏感性预测的当前进展”乳腺癌。