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

（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分离来自正常人肝的匀浆上清液，并转移到硝酸纤维素膜上。用辣根过氧化物酶标记的聚合人血清白蛋白（pHSA）与转移蛋白反应，检测到分子量为15 kD和14.8kD的两条带。这两种蛋白是pHSA的肝细胞受体（hepatocyte-pAR）。免疫荧光显微镜观察显示，正常人肝细胞pAR定位于细胞质和细胞膜。（2）肝病患者血清肝细胞-pAR及抗肝细胞-pAR抗体检测的临床意义：纯化的肝细胞-pAR包被于微孔板上，ELISA法检测血清抗肝细胞-pAR抗体。正常人血清中抗肝细胞pAR抗体均为阴性。HBsAg阳性患者。HBeAg阳性患者中56%， ...更多信息 58%的HBeAg阳性患者抗体阳性。抗人球蛋白抗体阳性患者。尤其是抗肝膜抗体阳性者，肝细胞pAR抗体阳性率高。部分HBeAg阳性患者血清中检测到肝细胞pAR，提示其与肝细胞损伤密切相关。（3）聚合人血清白蛋白（pHSA）的体内转归：将pHSA与胶体金结合，注入大鼠门静脉。用pHSA标记的胶体金对人肝活检标本进行灌注。肝窦内皮细胞和枯否细胞通过包被的小凹和囊泡摄取胶体金颗粒。需要进一步的研究来阐明pHSA和人肝细胞之间的相互作用。（4）粘附分子在肝细胞坏死中的意义：检测慢性B型肝炎患者肝活检组织中淋巴细胞相关抗原-1（LFA-1）和细胞间粘附分子-1（ICAM-1）的表达。LFA-1表达于浸润到片状坏死和局灶性坏死区域的淋巴细胞上。ICAM-1表达于肝细胞膜上，与HLA-1抗原相关。提示粘附分子在肝细胞坏死过程中起重要作用。少

(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