摘要
目的 研究人纤维介素基因或纤维介素/hfgl 2凝血酶原酶(简称hfgl 2)在重型肝炎、慢性肝炎、肝硬化患者肝组织及非配对患者外周血单个核细胞(PBMC)中的表达,分析其表达与患者临床表现和肝功能的关系,探讨hfgl 2检测对预测患者临床转归和预后的价值。方法 采用免疫组织化学法和免疫细胞化学法分别检测23例重型肝炎、13例慢性肝炎、14例肝硬化患者的肝组织和30例重型肝炎、10例慢性肝炎患者、10例健康对照者PBMC中的hfgl 2表达。应用多媒体彩色图文分析系统对hfgl 2的表达进行半定量分析。结果 23例重型肝炎患者肝组织中,21例可见hfgl 2表达,而在慢性乙型肝炎、乙型肝炎肝硬化患者肝组织中均无hfgl 2表达。肝组织中hfgl 2表达水平与血清总胆红素值呈正相关。30例重型肝炎患者中,28例外周血白细胞hfgl 2高表达,10例慢性乙型肝炎患者中有1例PBMC可检测到hfgl 2,健康对照组未检测到hfgl 2。PBMC中hfgl 2表达水平与血清总胆红素值呈正相关。结论 Hfgl 2的表达为重型肝炎所特有的现象,hfgl 2的表达与乙型肝炎患者病情的严重程度密切相关,减少和阻断其表达有可能为防治重型肝炎提供新的途径和方法。PBMC中hfgl 2的检测可能有助于重型肝炎的早期诊断和临床转归的判断。
Objective Viral hepatitis remains a major public health problem and the most common type of liver disease worldwide. There are an increasing number of patients with chronic hepatitis B who develop acute hepatitis on chronic condition (AOC) and die of acute hepatic failure both as a result of lack of understanding of the pathogenesis of the disease and lack of effective treatment. The hallmark of AOC is the extreme rapidity of the necromicroinflammatory process resulting in widespread or total hepatocellular necrosis in weeks or even days. Our previous studies have shown in an experimental animal model of fulminant viral hepatitis caused by murine hepatitis virus strain 3, the importance of macrophage activation, and expression of a unique gene mfgl 2 which encodes a serine protease capable of directly cleaving prothrombin to thrombin, resulting in widespread fibrin deposition within the liver and hepatocyte necrosis. The undergoing study in this report is designed to identify the role of hfgl 2 (human fibrinogen like protein 2)/fibroleukin in patients with viral hepatitis. Methods Liver tissues were obtained from 23 patients with AOC hepatitis B, and from 13 patients with inactive chronic hepatitis B (CHB) and 14 patients with chronic hepatitis B with cirrhosis during the year of 1995 to the end of 2001, Liver biopsies were performed within 30 min after the patients were diagnosed with death as a result of acute hepatic failure. Liver samples were also obtained from 4 liver donors as normal controls. In addition, Objective peripheral blood mononuclear cells(PBMC) were isolated from 30 patients (unpaired) with AOC hepatitis B and 10 patients with CHB during the May of 2001 to March of 2002 and l0 healthy volunteer as negative control. PBMCs were freshly isolated and smeared on slides and kept at -80℃ for further use. Histological sections were stained with hemotoxylin and eosin. A 169 bp of hfgl 2 cDNA probe and a polyclonal or monoclonal antibody against hfgl 2 were used to detect the expression of hfgl 2 mRNA and protein in liver samples as well as PBMC by immune histochemistry separately. Results Liver tissues from the patients with acute on chronic hepatitis had classical pathological features of acute necroinfammation. Hfgl 2 was detected by immune histochemistry in 21 of 23 patients (91.3%) in liver sections from patients with acute on CHB, while only 1 of 13 patients (7.7%) with CHB and cirrhosis and no evidence of active disease had hfgl 2 mRNA or protein expression. 28 of 30 patients (93.3%) with acute on CHB and 1 of 10 with CHB were detected with hfgl 2 expression in PBMC. There was no hfgl 2 expression in either the liver tissue or the PBMC from the normal donors. There was positive correlation of hfgl 2 expression and the severity of the disease displayed by the value of bilirubin and PT. Conclusion The molecular and cellular results reported here in patients with acute on chronic hepatitis and who died of acute hepatic failure correlates with previous report in 8 patients with fulminant hepatic failure (FHF) and mimic closely the changes observed in the murine model of fulminant viral hepatitis in which the pathogenesis of the disease has been studied in a stepwise fashion. This study further suggests that virus induced hfgl 2 prothrombinase/fibroleukin expression and the potent function of the protein it encodes plays a pivotal role in initiating acute severe hepatitis on the baseline of chronic hepatitis. The measurement of hfgl 2/fibroleukin expression in PBMC may serve as a useful marker to monitor the severity of disease in patients with the AOC hepatitis B and a target for therapeutic intervention.
出处
《中华肝脏病杂志》
CAS
CSCD
2004年第7期385-388,共4页
Chinese Journal of Hepatology
基金
国家自然科学基金(30170846)
国家杰出青年科学基金(30225040
30125019)
国家高技术研究发展计划(国科发财字[2002]469号)
