Basic and Clinical Studies on the Mechanism of Liver Cell Necrosis in Hepatits Type B

乙型肝炎肝细胞坏死机制的基础与临床研究

基本信息

批准号：
01570396
负责人：
TSUJI Takao
金额：
$ 1.34万
依托单位：
Okayama University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1989
资助国家：
日本
起止时间：
1989 至 1991
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-01570396/
关键词：
Hepatitis type B Viral Hepatitis Autoimmune Hepatitis Liver Membrane Antigen Anti-liver Membrane Antibody Mechanism of Liver Cell Necrosis Polymerized Human Serum Albumin Adhesion Molecules ICAMー1 LFAー1 肝細胞レセプタ-ウイルスレセプタ-

项目摘要

(1)Purification and Analysis of Hepatocyte Receptor for Polymerized Human Serum Albumin : Supernatant of homogenate from the normal human liver was separated by SDS-PAGE and transferred to nitrocellulose membrane. The transferred proteins were reacted with polymerized human serum albumin(pHSA)labeled with horseradish peroxidase and two bands with molecular weight of 15 kD and 14.8 kD were detected. The two proteins were hepatocyte receptors for pHSA(hepatocyte-pAR). Immunofluorescence microscopy using antibody to hepatocyte-pAR revealed hepatocyte-pAR was localized in cytoplasm and cell membrane of normal human hepatocytes.(2)Clinical Significance of Serum Hepatocyte-pAR and Antibody to Hepatocyte-pAR in Patients with Liver Diseases : Purified hepatocyte-pAR was coated on a microplate and serum antibody to hepatocyte-pAR was detected by ELISA. All sera from normal subjects were negative for antibody to hepatocyte-pAR. In patients with positive HBsAg. 56 % in HBeAg positive patients and … More 58 % in HBeAb-positive patients were positive for the antibody. Patients with positive HBsAb. especially with positive anti-liver membrane antibody, showed high frequency in hepatocyte-pAR antibody. Hepatocyte-pAR was detected in sera of some patients with positive HBeAg, suggesting the intimate relationship with liver cell injury.(3)Fate of Polymerized Human Serum Albumin(pHSA)in vivo : pHSA was conjugated with colloidal gold and injected into the portal vein of rats. Human liver biopsy specimen was also perfused with colloidal gold conjugated with pHSA. Colloidal gold particles were taken up by sinusoidal endothelial cells and Kupffer cells through coated pits and vesicles. Further studies are needed to elucidate the interaction between pHSA and human hepatocytes.(4)Significance of Adhesion Molecules in Liver Cell Necrosis : Adhesion molecules such as lymphocyte associate antigen-1(LFA-1)and intercellular adhesion molecules-1(ICAM-1)were examined in biopsy specimens from patients with chronic hepatitis type B. LFA-1 was expressed on lymphocytes infiltrating into the area such as piecemeal necrosis and focal necrosis. In such area, ICAM-1 was expressed on liver cell membrane in association with HLA class 1 antigen. These results suggest that adhesion molecule may play an important role in the process of liver cell necrosis. Less

(1)聚合人血清白蛋白的肝细胞受体的纯化和分析：通过SDS-PAGE分离正常人肝脏的匀浆上清液，并将转移的蛋白质与标记的聚合人血清白蛋白(pHSA)反应。检测到辣根过氧化物酶和两条分子量分别为15 kD和14.8 kD的条带，这两种蛋白是肝细胞受体。 pHSA(肝细胞-pAR)。使用肝细胞-pAR抗体进行免疫荧光显微镜检查显示肝细胞-pAR定位于正常人肝细胞的细胞质和细胞膜中。(2)肝细胞-pAR患者血清肝细胞-pAR和肝细胞-pAR抗体的临床意义。肝脏疾病：将纯化的肝细胞-pAR包被在微孔板上，并通过以下方法检测肝细胞-pAR的血清抗体： ELISA。在 HBsAg 阳性患者中，所有血清均呈阴性，而在 HBeAb 阳性患者中，尤其是 58% 呈阳性。抗肝膜抗体阳性，表明在一些HBeAg阳性患者的血清中检测到肝细胞-pAR抗体的频率较高。 (3)聚合人血清白蛋白(pHSA)的体内命运：将pHSA与胶体金结合并注射到大鼠门静脉中，同时用胶体金结合物灌注人肝活检标本。胶体金颗粒通过包被的小凹和囊泡被窦内皮细胞和库普弗细胞吸收。阐明pHSA与人类肝炎之间的相互作用。(4)粘附分子在肝细胞坏死中的意义：在活检中检查淋巴细胞相关抗原-1(LFA-1)和细胞间粘附分子-1(ICAM-1)等粘附分子慢性乙型肝炎患者标本中，LFA-1表达于片状坏死、局灶等浸润区域的淋巴细胞上。在该区域，ICAM-1在肝细胞膜上表达，并与HLA 1类抗原相关。这些结果表明，粘附分子可能在肝细胞坏死过程中发挥重要作用。