丁型肝炎患者肝组织中Fas/FasL和肝细胞凋亡表达

目的探讨Fas/FasL及肝细胞凋亡在丁型肝炎患者肝组织中的表达及作用.方法采用免疫组化双重染色和TUNEL技术,检测79例丁肝患者肝组织HDAg,Fas/FasL表达,以及肝细胞凋亡,以54例乙型肝炎作对照.结果Fas以肝细胞浆表达为主,HDAg以核浆型表达为主,二抗原表达及分布有相关性.FasL主要表达在浸润淋巴细胞浆、肝细胞核,与HDAg表达及分布不全一致.HDAg,Fas/FasL和肝细胞凋亡在各型肝炎中的表达强度有显著性差别意义.结论HDV感染可诱导肝细胞Fas/FasL表达,增强Fas/FasL途径介导的肝细胞凋亡,这一机制在丁型肝炎发病机制中可能有一定的重要作用.     

An aggressive nasal lymphoma accompanied by high levels of soluble Fas ligand

Fas ligand (FasL), either in the membrane bound form or soluble form, has cytotoxic activity against Fas-expressing cells. We report a case of nasal lymphoma accompanied by liver damage and pancytopenia. The serum level of soluble FasL (sFasL) was very high on admission, but rapidly decreased to normal levels after chemotherapy for lymphoma. Liver damage and pancytopenia also improved with the decrease in serum sFasL. Since Fas is expressed on both hepatocytes and haemopoietic cells, these facts suggest that FasL was expressed on lymphoma cells and directly associated with pathogenesis of liver damage and pancytopenia through its cytotoxic activity.     

An aggressive nasal lymphoma accompanied by high levels of soluble Fas ligand

Fas ligand (FasL), either in the membrane bound form or soluble form, has cytotoxic activity against Fas-expressing cells. We report a case of nasal lymphoma accompanied by liver damage and pancytopenia. The serum level of soluble FasL (sFasL) was very high on admission, but rapidly decreased to normal levels after chemotherapy for lymphoma. Liver damage and pancytopenia also improved with the decrease in serum sFasL. Since Fas is expressed on both hepatocytes and haemopoietic cells, these facts suggest that FasL was expressed on lymphoma cells and directly associated with pathogenesis of liver damage and pancytopenia through its cytotoxic activity.     

In situ investigation of Fas/FasL expression in chronic hepatitis B infection and related liver diseases

To evaluate Fas/FasL expression in hepatitis B virus-related chronic liver disease, liver biopsies from 44 such cases were studied immunohistochemically. FasL was detected in the infiltrating lymphocytes and both FasL and Fas were found in the hepatocytes. The Fas and FasL-positive cells were mostly found at the advancing edges of interphase hepatitis, and Fas/FasL expression was closely correlated with the inflammatory activity. Unexpectedly, FasL was also expressed in liver cirrhotic nodules, particularly in those with hepatocellular carcinoma with or without inflammation. These results suggest that the factors which induce hepatocyte transformation might also trigger FasL expression and promote FasL/Fas-mediated apoptosis.     

乙型肝炎患者肝组织中Fas及Fas配体的表达

为探讨乙型肝炎肝组织中Ｆａｓ及Ｆａｓ配体（ＦａｓＬ）表达和分布的相互关系，采用免疫组织化学技术，以兔抗－Ｆａｓ及兔抗－ＦａｓＬ多克隆抗体，对６０例急性轻型肝炎、慢性活动性肝炎及活动性肝硬化患者石蜡包埋肝组织中的Ｆａｓ及ＦａｓＬ进行了检测。Ｆａｓ及ＦａｓＬ的检出率分别为７６．７％（４６／６０）及７０．０％（４２／６０），Ｆａｓ于肝细胞胞浆内表达，ＦａｓＬ多在肝组织中浸润的淋巴细胞胞浆内表达（３４／４２，８０．９％），也见于肝细胞胞浆中（２５／４２，５９．５％）。ＦａｓＬ阳性淋巴细胞主要分布于汇管区，肝小叶内很少见，ＦａｓＬ阳性肝细胞的分布类同Ｆａｓ阳性肝细胞；在急性轻型肝炎，阳性细胞多在小叶内弥散分布，在慢性活动性肝炎和活动性肝硬化则更多集聚于碎屑样坏死灶和假小叶的周边。免疫组化双标记染色显示，Ｆａｓ及ＦａｓＬ可在同一或不同肝细胞胞浆内表达，但多分布于同一区域。本研究在正常肝组织中未查见ＦａｓＬ的表达，提示在病理状态下，肝细胞才出现ＦａｓＬ的表达。Ｆａｓ及ＦａｓＬ在肝组织中的分布表明，Ｆａｓ－ＦａｓＬ系统在乙型病毒性肝炎肝细胞损伤中起着重要的作用。     

丙型肝炎肝组织Fas配体的免疫组化研究

以免疫组化法研究Ｆａｓ配体（Ｌ）在丙型肝炎病人肝组织内的表达情况，发现在碎屑样坏死（ＰＮ）、桥样坏死（ＢＮ）及灶性坏死性浸润的单个核细胞有明显的ＦａｓＬ表达，阳性率为４１．２％。ＦａｓＬ阳性细胞与肝脏炎症分级程度密切相关，Ｇ０－１组与Ｇ３－４组比较有显著差异。ＦａｓＬ在肝细胞内亦有表达，表达程度似与炎症程度有关。不论是Ｆａｓ或ＦａｓＬ在慢性丙肝中的表达均较急性肝炎显著，提示在病程慢性化中可能起着重     

乙型肝炎患者肝组织中FasL的检测

目的探讨乙型病毒性肝炎患者肝组织中Ｆａｓ配体（ＦａｓＬ）的表达与分布及其与Ｆａｓ抗原的相互关系。方法应用免疫组织化学双标记技术，以免抗－ＦａｓＬ及兔抗一Ｆａｓ多克隆抗体，对４６例慢性肝炎及活动性肝硬化患者石蜡包埋肝组织中的ＦａｓＬ及Ｆａｓ进行了检测。结果Ｆａｓ及ＦａｓＬ的检出率分别为７１．７％（３３／４６）及６５．２％（３０／４６），Ｆａｓ于肝细胞胞浆内表达；ＦａｓＬ多在肝组织中浸润的淋巴细胞胞浆内表达（２３／３０，７ｔｉ，７％），也见于肝细胞浆中（２０／３０，６６．７％）。ＦａｓＬ阳性淋巴细胞主要分布于汇管区，肝小叫内很少见，ＦａｓＬ阳性肝细胞的分布类同Ｆａｓ阳性肝细胞：即多集聚于碎屑样坏死灶和假小叶的周边。免疫组化双标记染色显示Ｆａｓ及ＦａｓＬ可在同一或不同肝细胞胞浆内表达，但多分布于同一区域。结论病毒性肝炎肝组织中ＦａｓＬ的分布与Ｆａｓ抗原密切相关，Ｆａｓ／ＦａｓＬ系统在肝细胞损伤中起了重要的作用，肝细胞内ＦａｓＬ表达的意义尚待进一步研究。     

血吸虫病小鼠肝纤维化肝组织中Fas/FasL的表达

目的 探讨Fas／FasL在血吸虫病肝纤维化肝组织中的表达及意义。 方法 采用免疫组化技术检测血吸虫病小鼠肝纤维化肝组织中Fas／FasL的表达,并对照经吡喹酮、已酮可可碱治疗前后其表达的变化,分析其意义。 结果Fas以肝细胞浆表达为主,FasL主要在浸润淋巴细胞浆、肝细胞核表达。经吡喹酮及高剂量已酮可可碱治疗后其肝组织Fas／FasL表达明显减少(P<0.01)。 结论 血吸虫感染可诱导肝细胞Fas／FasL的表达,增强Fas／FasL途径介导的肝细胞凋亡,这一机制在血吸虫病肝纤维化的发病中可能起重要作用。吡喹酮及高剂量已酮可可碱可明显下调Fas／FasL的表达,抑制肝细胞凋亡,改善肝功能。     

丁型肝炎病人肝组织HDAg和 Fas表达及其相互关系

目的 研究丁型肝炎病人肝组织Fas和HDAg表达及其相互关系,了解Fas在HDV致病机制中的作用。方法 应用免疫组化单、双标记技术,检测48例丁型肝炎病人肝组织Fas、HDAg表达,以54例乙型肝炎病人肝组织Fas表达作对照。结果 Fas以肝细胞浆表达为主,HDAg以肝细胞核和胞浆表达为主,二抗原表达及分布呈一致性。各型肝炎中Fas和HDAg表达有统计学差别,两种抗原的分布和表达强度与肝细胞炎症活动和病理损害程度相关。结论 Fax和HDAg表达及分布密切相关,与肝组织炎症活动和病理损害相关,此提示HDV感染可诱导肝细胞表达Fas,Fas及其所介导的肝细胞凋亡在HDV致病机制中可能有重要作用。     

HDV/HBV感染树鼩肝组织Fas/FasL表达与肝细胞凋亡

目的 探讨HDV感染树鼩肝组织中Fas/FasL表达与HDV感染之间的关系,以及Fas/FasL在丁型肝炎肝细胞凋亡中的作用。 方法 采用免疫组化和原位杂交技术对45份HDV感染树鼩肝组织中HDAg,Fas/FasL和Fas/FasL mRNA的表达进行了检测;应用原位末端标记技术对肝细胞凋亡进行了检测;并应用免疫组化双重染色对HDAg,Fas/FasL的表达以及肝细胞凋亡进行了检测。 结果 45份肝组织中有39份可检出Fas/FasL(阳性率87％),有41份可检出凋亡细胞(阳性率91％),HDAg表达与Fas/FasL表达之间有显著相关性(X_1~2=29.2,X_2~2=27.9,P<0.05),HDAg表达越强,Fas和FasL表达也越强,凋亡在HDAg阳性和阴性细胞中均可发生,以HDAg阳性细胞发生为主,Fas/FasL表达与肝细胞凋亡之间有显著相关性(X_1~2=35.1,X_2~2=40.2,p<0.05),Fas和FasL表达越强,凋亡阳性细胞越多。 结论 丁型肝炎病毒感染和未感染的肝细胞均可发生凋亡,但凋亡只在少数细胞发生;肝细胞内的病毒抗原表达可诱导Fas/FasL的表达;Fas/FasL肝细胞凋亡中起重要作用。     

Fas、FasL在慢性乙型肝炎肝组织中的表达

探讨Fas、FasL在慢性乙型肝炎肝组织中的表达特点。对 30例慢性乙型肝炎患者进行肝穿刺肝组织病理检查及免疫组化法检测肝组织中的Fas/FasL表达强度。 (1)Fas、FasL在肝组织肝细胞膜及胞浆均有不同程度表达 ,其表达阳性细胞主要位于汇管区小叶内及周边碎屑状坏死区 ,呈弥漫分布 ,在其周围浸润的淋巴细胞上多为FasL阳性细胞 ,亦可见到Fas阳性细胞。 (2 )肝组织中Fas、FasL表达随着慢性肝炎程度即炎症和纤维化程度加重而加强 (P均 <0 0 0 1)。Fas-FasL系统介导的细胞凋亡 ,确实参与了慢性乙型肝炎的发病机制 ,且在慢性乙型肝炎的发生发展中起重要作用。     

Severe necroinflammatory reaction caused by natural killer cell-mediated Fas/Fas ligand interaction and dendritic cells in human hepatocyte chimeric mouse

The necroinflammatory reaction plays a central role in hepatitis B virus (HBV) elimination. Cluster of differentiation (CD)8-positive cytotoxic T lymphocytes (CTLs) are thought to be a main player in the elimination of infected cells, and a recent report suggests that natural killer (NK) cells also play an important role. Here, we demonstrate the elimination of HBV-infected hepatocytes by NK cells and dendritic cells (DCs) using urokinase-type plasminogen activator/severe combined immunodeficiency mice, in which the livers were highly repopulated with human hepatocytes. After establishing HBV infection, we injected human peripheral blood mononuclear cells (PBMCs) into the mice and analyzed liver pathology and infiltrating human immune cells with flow cytometry. Severe hepatocyte degeneration was observed only in HBV-infected mice transplanted with human PBMCs. We provide the first direct evidence that massive liver cell death can be caused by Fas/Fas ligand (FasL) interaction provided by NK cells activated by DCs. Treatment of mice with anti-Fas antibody completely prevented severe hepatocyte degeneration. Furthermore, severe hepatocyte death can be prevented by depletion of DCs, whereas depletion of CD8-positive CTLs did not disturb the development of massive liver cell apoptosis. CONCLUSION: Our findings provide the first direct evidence that DC-activated NK cells induce massive HBV-infected hepatocyte degeneration through the Fas/FasL system and may indicate new therapeutic implications for acute severe/fulminant hepatitis B.     

38例急性乙型病毒性肝炎肝组织中Fas抗原和Fas配体的表达研究

目的:探讨Fas-FasL系统在急性病毒性肝炎发病中的作用.方法:采用免疫组织化学技术.结果:38例急性乙型肝炎肝组织中的Fas和FasL表达的检出率分别为55.3%和66.5%;Fas和FasL的表达强度与患者的年龄、性别和血清A、A/G和PTA无关,而与血清ALT和TBIL水平有关.较重型(伴桥样坏死)的急性乙型肝炎其Fas和FasL表达比轻型急性乙型肝炎要强.结论:由CTL-Fas-FasL系统介导的肝细胞凋亡在急性乙型肝炎的发病中起了较重要的作用.     

慢性乙型肝炎和肝病中Fas和FasL表达的原位研究

目的了解慢性乙型肝炎和肝病时介导凋亡的Ｆａｓ／ＦａｓＬ表现。方法以免疫组化法原位检查各种慢性肝病４５例的活检肝组织。结果肝内浸润的淋巴细胞中检出ＦａｓＬ，肝细胞Ｆａｓ／ＦａｓＬ表达与炎症活动性一致，多分布在界面性炎症区。肝细胞表达ＦａｓＬ，可能也发挥细胞毒效应。结论新的发现是ＦａｓＬ可在肝细胞结节和肝癌细胞表达，提示有细胞毒效应的ＦａｓＬ在ＨＢ相关慢性肝病中有发病学意义。肝癌细胞中未检出Ｆａｓ而检出ＦａｓＬ，可能是一种肿瘤逃避免疫攻击的机制     

肝细胞凋亡与慢性乙型肝炎病毒携带者肝组织病变的关系

目的 观察慢性HBV携带者（ASC）肝组织凋亡指数及Fas、FasL的表达,探讨肝细胞凋亡在ASC肝组织病变中的作用.方法 选取HBV携带者患者120例,并进行肝组织活检.采用原位末端转移酶标技术进行肝细胞凋亡情况检测,采用免疫组织化学技术检测肝组织中的Fas、FasL.结果 120例ASC肝组织完全正常者仅占11％,轻度肝炎组占59％,而中、重度肝炎组占30％.ASC患者的凋亡指数（AI）随着炎症程度加重而升高,正常组为1.50±1.04,而中度组、重度组分别为11.33 ±1.51和17.67 ±6.42,各组间比较差异有统计学意义（P＜0.05）.肝组织免疫组化结果显示,Fas、FasL表达程度随炎症活动度加重而增强,尤其在汇管区周围碎屑状坏死区及周边小叶更明显,正常组基本无表达,而轻度组以无表达或阳性表达为主,仅见于界面炎症区的肝细胞及淋巴细胞上;中、重度肝炎组Fas、FasL则均为阳性及强阳性表达,除界面炎症区外,肝小叶中也弥漫分布.Fas强阳性表达率在中、重度组分别为34.8％和69.2％,FasL的强阳性表达率分别为30.4％和53.8％,均明显高于正常组和轻度组（P＜0.05）,同时中、重度组间比较差异亦有统计学意义,肝组织中Fas、FasL表达程度随炎症活动度加重而增强,两者间明显相关.结论 肝细胞凋亡在ASC肝组织病变的发生发展中可能起到重要作用.     

Fas、FasL在慢性乙型肝炎患者肝细胞和外周血淋巴细胞上的表达

探讨慢性乙型肝炎患者肝细胞及外周血淋巴细胞上Fas、FasL的表达与乙型肝炎病毒（HBV）复制水平及肝组织炎症程度的关系。对30例慢性乙型肝炎患者进行肝穿刺肝组织病理检查及免疫组化SP法检测肝细胞中Fas、FasL表达强度。并采用单克隆抗体经流式细胞仪检测外周血淋巴细胞上Fas、FasL表达阳性百分率;同时,采用荧光实时标记法测定血清中病毒复制指标HBV DNA水平;采用Beckman全自动生化分析仪检测肝功能并研究其相关性。慢性乙型肝炎患者肝细胞中Fas、FasL表达强度随肝组织炎症活动度加重而增强（P均〈0．001）,与白蛋白及丙氨酸转移酶（ALT）呈负相关（P〈0．005,P〈0．001）,与球蛋白、总胆红质无明显相关性（P均〉0．05）。外周血淋巴细胞上Fas、FasL的表达阳性率与血清中肝炎病毒复制指标HBV DNA水平呈正相关;与慢性肝炎分度无明显的相关性。乙型肝炎病毒感染可诱导肝细胞及外周血淋巴细胞上Fas、FasL的表达。肝细胞Fas、FasL的表达随炎症活动度加重而表达增强,但其介导的凋亡并不引起肝细胞炎症损伤即ALT活性并不升高,相反减少;同时在某种程度上影响白蛋白的合成。提示Fas—FasL介导的肝细胞凋亡是以非细胞损伤方式参与慢性乙型肝炎的发病过程。同时,随血清HBV DNA水平增高,外周血淋巴细胞上Fas、FasL的表达亦增强,淋巴细胞的凋亡因而增多,是导致乙型肝炎慢性化的原因之一。由此可见,Fas、FasL介导的凋亡参与了慢性乙型肝炎的发病机制,且在慢性乙型肝炎的发生发展中起重要作用。     

Involvement of Fas system-mediated apoptosis in pathogenesis of viral hepatitis

Enhanced Fas system-mediated hepatocyte apoptosis is observed in hepatitis C virus (HCV)- and hepatitis B virus (HBV)-associated chronic liver diseases. In these forms of viral hepatitis, liver-infiltrating lymphocytes that recognize the viral antigen on hepatocytes become activated and express cytolytic Fas ligand (FasL) molecules. In contrast, hepatocytes exhibit enhanced Fas expression and become susceptible to FasL-mediated death. Augmentation of the Fas system has also been observed in other liver diseases and biliary disorders. Moreover, the Fas system is involved in removing aged hepatocytes. Thus, Fas-mediated apoptosis plays an important role in several liver diseases and in maintaining normal liver homeostasis.