摘要
目的探讨肝硬化患者糖代谢紊乱与肝细胞胰岛素受体(IR)和酪氨酸蛋白激酶(TPK)表达,以及胰腺细胞HBV DNA阳性表达的关系。方法 应用地高辛素标记HBV DNA s 577 bp探针原位杂交技术,检测12例血清HBV标志物阳性的肝炎肝硬化患者肝、胰组织内的HBV DNA;应用图象分析系统对该12例肝细胞IR和TPK,进行抗胰岛素受体和酪氨酸蛋白激酶抗体免疫组化标记物定量测定。用免疫荧光组织化学双重染色技术,激光共聚焦扫描显微镜观察HBsAg和lR。肝、胰组织活检前,常规进行静脉葡萄糖耐量试验。结果 肝细胞HBV DNA阳性11例(11/12),糖耐量试验异常(IGT)7例中肝细胞HBV DNA均阳性(7/7),糖耐量试验正常(NGT)5例中HBV DNA阳性4例(4/5)。胰腺细胞阳性8例(8/12),IGT7例中胰腺细胞HBV DNA均阳性(7/7),NGT5例中仅1例HBV DNA阳性(1/5),IGT组与NGT组胰腺细胞检出HBV DNA差异有显著意义(P<0.05)。IGT的肝硬化患者肝细胞内IR和TPK表达较NGT的肝硬化患者差异有非常显著意义(tIR=3.617 P<0.O1,tTPK=20.143P<0.01)。肝细胞内IR与TPK表达量高度相关,r=0.82597(P<0.01)。免疫荧光组织化学双重染色显示肝细胞和胰岛细胞IR阳性处有HBsAg存在。结论HBV不仅能侵害肝细胞,也能直接侵害胰岛细胞。
Objective To investigate relationship between glucose metabolic disorders and expression of insulin receptor (IR) and tyrosine protein kinase(TPK) in posthepatitic cirrhosis hepatocyte and HBV DNA expression in pancreatic cells. Methods To detect HBV DNA in paraffin-embeded pancreatic and hepatic tissues from 12 posthepatitic cirrhosis patients with positive serum HBV markers by using in situ hybridization (ISH) with a digoxigenin labelled probe.The amount of IR and TPK have been evaluated by immunohistochemical quantitative analysis using image analyzer in hepatocyte of 12 patients positive for HBV markers with posthepatitic cirrhosis in serum. Immunofluorescent histochemical double staining technique was used. HBsAg and IR were observed under confocal laser scanning microscope. Results Eleven of 12 cirrhosis patients' hepatocytes were HBV DNA positive, including 7 patients (7/7) with impaired glucose tolerance (IGT) and 4 patients (4/5) with normal glucose tolerance (NGT). Eight of 12 pancreatic cells were HBV DNA positive, including 7 patients (7/7) with IGT, but only one patient (1/5) with NGT. HBV DNA was found positive in pancreatic cells in signifficantly more subjects in IGT group than in NGT group ( P < 0.01) . IR and TPK amount in hepatocyte of IGT was signifficantly less than that of NGT patients with posthepatitic cirrhosis (P < 0.01) . IR amount was closely related to the TPK in cirrhosis hepatocyte r=0.82597( P < 0.01) . HBV DNA was mainly localized in the nuclei of hepatocyte and pancreatic acinar and islet cells. Immunofluorescent histochemical double-staining showed that HBsAg was partly localized in the IR positive areas of hepatocytes and pancreatic islet cells. Conclusion HBV can invade acinar cells of pancreas and islet cells, which might be a direct cause of insulin-dependent diabetes mellitus-like disorder and insulin absence after HBV infection. Decrease of IR and TPK might be the main cause of noninsulin-dependent diabetes mellitus-like disorder after having hepatitis or posthepatitic cirrhosis.
出处
《中华实验和临床病毒学杂志》
CAS
CSCD
北大核心
2003年第4期372-374,F003,共4页
Chinese Journal of Experimental and Clinical Virology
