诱骗受体3在HBeAg阴性的慢性乙型肝炎患者中的表达及意义

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侯彦强, 梁冬雨, 彭亮, 娄晓丽, 刘涛, 张鲁榕. 诱骗受体3在HBeAg阴性的慢性乙型肝炎患者中的表达及意义. 2013, 28(1): 37-39
HOU Yanqiang, LIANG Dongyu, PENG Liang, LOU Xiaoli, LIU Tao, ZHANG Lurong. Expression and significance of decoy receptor 3 in HBeAg negative chronic hepatitis B patients. Labratory Medicine, 2013, 28(1): 37-39 复制到剪切板

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诱骗受体3在HBeAg阴性的慢性乙型肝炎患者中的表达及意义

侯彦强¹, 梁冬雨¹, 彭亮¹, 娄晓丽¹, 刘涛¹, 张鲁榕²

1. 上海市松江区中心医院中心实验室,上海 201600

2. 美国佛罗里达大学肿瘤研究中心,美国佛罗里达 32610

作者简介：侯彦强,男,1971年生,博士,副主任技师,主要从事临床感染免疫基础及检验研究。

基金:基金项目上海市科委科研课题资助项目（12ZR142800）;

摘要

目的

探讨诱骗受体3(DcR3)在乙型肝炎病毒(HBV)e抗原 (HBeAg) 阴性的慢性乙型肝炎(CHB)患者中的表达及意义。

方法

采用酶联免疫吸附试验(ELISA)检测80例血清HBeAg阴性的CHB患者和96名正常人血清DcR3;全自动生化分析仪检测丙氨酸氨基转移酶(ALT);聚合酶链反应(PCR)检测HBV DNA。

结果

HBeAg阴性的CHB患者血清DcR3水平[1.58(0.483.23) ng/mL]明显高于正常对照组[0.83(0.162.32) ng/mL,P<0.000 1)]。HBeAg阴性的CHB患者血清DcR3水平与HBV DNA和ALT水平呈正相关(r=0.704、0.732,P均<0.000 1) 。

结论

DcR3在HBeAg阴性的CHB患者中高表达,并与HBV DNA和ALT强相关,具有潜在的作为HBeAg阴性CHB患者诊断、治疗观测的新指标的应用价值。

关键词: 诱骗受体3; 慢性乙型肝炎; 丙氨酸氨基转移酶; DNA

中图分类号:R446.62 文献标志码:A 文章编号:1673-8640(2013)01-0037-03

Expression and significance of decoy receptor 3 in HBeAg negative chronic hepatitis B patients

HOU Yanqiang¹, LIANG Dongyu¹, PENG Liang¹, LOU Xiaoli¹, LIU Tao¹, ZHANG Lurong²

1. Department of Central Laboratory,Songjiang Central Hospital,Shanghai 201600,China

2. Shands Cancer Center,University of Florida,Florida 32610,U.S.A

Abstract

Objective

To investigate the expression and significance of decoy receptor (DcR3) in hepatitis B virus (HBV) e-antigen (HBeAg) negative chronic hepatitis B (CHB) patients.

Methods

Serum DcR3 levels were measured by enzyme-linked immunosorbent assay (ELISA) in 80 HBeAg negative CHB patients and 96 healthy subjects. Alanine aminotransferase (ALT) was determined by automatic analyzer. Serum HBV DNA was determined by polymerase chain reaction (PCR).

Results

DcR3 level in HBeAg negative CHB patients [1.58(0.48-3.23)ng/mL]was significantly higher than that in healthy controls [0.83(0.16-2.32) ng/mL,P<0.000 1]. DcR3 level was correlated positively with HBV DNA (r=0.704,P<0.000 1) and ALT (r=0.732,P<0.000 1).

Conclusions

DcR3 has high expression and correlates with HBV DNA and ALT in HBeAg negative CHB patients,and DcR3 may serve as a diagnosis and treatment monitoring indicator for HBeAg negative CHB patients.

Keyword: Decoy receptor 3; Chronic hepatitis B; Alanine aminotransferase; DNA

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引言

近年乙型肝炎病毒(HBV)e抗原(HBeAg)阴性的慢性乙型肝炎(chronic hepatitis B,CHB)患者比例逐年增加。有报道亚洲CHB患者中,HBeAg阴性的乙型肝炎在日本占19%,在中国大陆占21%40%,中国台湾为20%,香港为11%,印度为16%^{[ 1]}。这类CHB患者由于HBeAg为阴性,常被认为病毒活动性低而忽视治疗,但许多HBeAg阴性的CHB患者病毒仍处于活动状态。HBV DNA检测是目前最常用的评估病毒活动情况的实验室指标,但该项目检测需要昂贵的仪器设备、试剂,需要专业培训的技术人员,这在一定程度上限制了该项目的广泛应用。诱骗受体3(decoy receptor 3,DcR3)是新近发现的一种缺乏穿膜结构域的可溶性肿瘤坏死因子受体超家族成员,能够竞争性结合凋亡相关因子配体(FasL)、肿瘤坏死因子超家族第14个成员(LIGHT)和肿瘤坏死因子配体相关分子1A(TL1A)3种肿瘤坏死因子超家族配体,在调控细胞增殖、分化和凋亡中发挥重要作用。有研究发现,DcR3具有潜在的作为炎性疾病的多功能免疫调节分子和标志物的临床应用价值,其在许多炎症性疾病中表达上调,如细菌性感染、肾炎、类风湿性关节炎、急性溃疡性结肠炎、阑尾炎等^{[ 2, 3]}。我们通过检测HBeAg阴性的CHB患者血清DcR3的表达水平,探讨DcR3是否具有作为HBeAg阴性的CHB标志物的临床应用价值。

材料和方法

一、研究对象及材料

选取2009年2月至2010年11月上海市松江区中心医院传染科就诊的乙型肝炎患者。收集80例HBeAg阴性CHB患者和96名正常人血清样本,-80 ℃保存备用。CHB患者诊断符合《慢性乙型肝炎防治指南》(2010年)中病毒性肝炎的诊断标准,排除脂肪肝、酒精性肝病、肥胖症、肝癌、丙型肝炎、丁型肝炎、人类免疫缺陷病毒(HIV)感染、自身免疫性肝病及其他肝病。HBeAg阴性CHB组中男57例,女23例,年龄(44±12)岁,丙氨酸氨基转移酶(ALT)(78.5±49.3)U/L,HBV DNA为(4.5±1.9)log拷贝/mL。正常对照组男65名,女31名,年龄(41±15)岁,ALT为(13.9±9.7)U/L。DcR3单克隆抗体和生物素标记的DcR3抗体由美国佛罗里达大学肿瘤研究中心张鲁榕教授惠赠。

二、方法

1. ALT和HBV DNA分析

ALT测定采用日立7600全自动生化分析仪,HBV DNA定量检测采用罗氏聚合酶链反应(PCR)诊断系统(Roche PCR diagnostic system),操作均严格按仪器及试剂说明书进行。

2. 血清DcR3检测

本研究利用自行建立的血清DcR3检测方法对各组样本进行检测。具体步骤如下:DcR3 单克隆抗体包被酶联免疫吸附试验(ELISA)板过夜,次日洗板,5%牛血清白蛋

白(BSA)封阻。加入不同浓度DcR3纯标准品(0、0.25、0.5、1、2、4、8和16 ng/mL)和各待测样本,室温2 h,充分洗板;加入生物素标记的DcR3单克隆抗体,室温2 h,洗板;加抗生物素蛋白链菌素-辣根过氧化物酶(streptavidin-HRP)孵育30 min,洗板;加四甲基联苯胺(TMB)避光反应10 min,加终止液终止反应,酶标仪上450 nm测量吸光度( A)值,由标准曲线求出各血清样本浓度。

三、统计学方法

采用SPSS 13.0软件进行数据分析,数据用中位数(范围)表示,采用非参数Mann-Whitney方法比较各组间差异,采用Spearman方法进行相关性统计分析, P<0.05表示差异有统计学意义。

一、血清DcR3检测

HBeAg阴性CHB组DcR3为1.58(0.483.23) ng/mL,正常对照组DcR3为0.83(0.162.32) ng/mL;2组比较差异有统计学意义( P<0.000 1)。见图1。

	Figure Option View Download New Window
	图1 血清DcR3水平

二、HBeAg阴性CHB患者中血清DcR3与ALT和HBV DNA表达相关性分析

HBeAg阴性CHB患者血清DcR3水平与HBV DNA和ALT水平呈正相关( r值分别为0.704、0.732, P均<0.000 1)。见图2。

	Figure Option View Download New Window
	图2 DcR3与ALT、HBV DNA 的相关性注:(a)DcR3与ALT的相关性( r=0.732, P<0.000 1);(b)DcR3与HBV DNA的相关性( r=0.704, P<0.000 1)

讨论

HBeAg阴性CHB在CHB自然史中处于较晚期阶段,患者年龄往往较HBeAg阳性CHB患者大,平均为40岁左右,以男性为主。HBeAg阴性CHB患者肝脏组织学损害明显重于HBeAg阳性CHB患者,发病率近年来不断上升。抗病毒是治疗CHB的有效手段,通常将HBeAg转为HBV e抗体(抗HBe)作为评价治疗效果的标志之一,但HBeAg阴性CHB患者不可能出现这种血清学转换。ALT和HBV DNA是目前评估肝脏损伤和病毒活动状况的最常用指标,但由于HBV DNA成本较贵、需要专业技术人员、操作复杂等原因,其广泛应用受到限制,尤其是在许多低收入国家和地区。

本研究结果显示,HBeAg阴性CHB组血清DcR3[1.58(0.483.23) ng/mL]明显高于正常对照组[0.83(0.162.32) ng/mL, P<0.000 1];HBeAg阴性CHB患者血清DcR3水平与HBV DNA和ALT水平呈正相关( r=0.704、0.732, P均<0.000 1)。因此,DcR3的表达水平在一定程度上可以反映ALT和HBV DNA的变化,这具有重要的临床实际应用价值。

DcR3在HBV感染中的作用目前尚不清楚。已知炎性免疫反应在HBV感染中发挥重要作用。DcR3表达水平的升高,可能是机体对病毒感染所致炎症代偿性保护反应的结果。有研究者利用转基因技术研究发现,DcR3能影响巨噬细胞的定向分化,从而抑制机体免疫应答,发挥抗炎作用^{[ 4, 5]}。在一些炎症性疾病中,DcR3在患者体内都有不同程度的表达上调。Kim等^{[ 6]}研究发现,急性阑尾炎患者的阑尾上皮细胞中高表达DcR3,体外利用脂多糖(LPS)刺激肠上皮细胞系SW480、SW620和HT29,发现DcR3释放明显增加。在急性溃疡性结肠炎发病过程中,DcR3的表达与炎症程度密切相关,在炎症高发期患者外周血DcR3呈现高水平表达,经有效治疗后,外周血DcR3含量明显降低^{[ 7]}。Yang等^{[ 8]}研究表明,肝癌患者血清中DcR3明显升高,且血清中含量与肝硬化、肿瘤转移和复发相关,与肝癌组织中DcR3表达和分布高度相关,提示DcR3在原发性肝癌的发病、转移和复发过程中发挥了重要作用。

综上所述,DcR3具有作为HBeAg阴性CHB标志物的潜在应用价值,可作为除HBV DNA和ALT之外的一项辅助实验指标,或作为HBV DNA的替代指标应用于临床。但DcR3的具体临床价值还有待于进一步深入研究。

The authors have declared that no competing interests exist.

参考文献

View Option

[1]	Funk ML, Rosenberg DM, Lok AS. World-wide epidemiology of HBeAg-negative chronic hepatitis B and associated precore and core promoter variants[J]. J Viral Hepat, 2002, 9(1): 52-61. [本文引用:1] [JCR: 3.082]
[2]	娄晓丽, 梁冬雨, 侯彦强. 诱骗受体3在疾病检测中的发展[J]. 中华检验医学杂志, 2011, 34(10): 877-880. [本文引用:1]
[3]	Lin WW, Hsieh SL. Decoy receptor 3: a pleiotropic immunomodulator and biomarker for inflammatory diseases, autoimmune diseases and cancer[J]. Biochem Pharmacol, 2011, 81(7): 838-847. [本文引用:1] [JCR: 4.576]
[4]	Chang YC, Hsu TL, Lin HH, et al. Modulation of macrophage differentiation and activation by decoy receptor 3[J]. J Leukoc Biol , 2004, 75(3): 486-494. [本文引用:1]
[5]	Chang YC, Chen TC, Lee CT, et al. Epigenetic control of MHC class Ⅱ expression in tumor-associated macrophages by decoy receptor 3[J]. Blood, 2008, 111(10): 5054-5063. [本文引用:1] [JCR: 9.06]
[6]	Kim S, Fotiadu A, Kotoula V. Increased expression of soluble decoy receptor 3 in acutely inflamed intestinal epithelia[J]. Clin Immunol, 2005, 115(3): 286-294. [本文引用:1] [JCR: 3.771]
[7]	Bamias G, Kaltsa G, Siakavellas SI, et al. High intestinal and systemic levels of decoy receptor 3 (DcR3) and its ligand TL1A in active ulcerative colitis[J]. Clin Immunol, 2010, 137(2): 242-249. [本文引用:1] [JCR: 3.771]
[8]	Yang M, Chen G, Dang Y, et al. Significance of decoy receptor 3 in sera of hepatocellular carcinoma patients[J]. Ups J Med Sci, 2010, 115(4): 232-237. [本文引用:1]

2002

3.082

0.0

. 2002, 9(1):52-61

World-wide epidemiology of HBeAg-negative chronic hepatitis B and associated precore and core promoter variants

M. L. Funk 1,2 ,D. M. Rosenberg 3 andA. S. F. Lok 4

1 Department of Epidemiology, University of North Carolina, Chapel Hill, North Carolina, USA, 2 Worldwide Epidemiology Department, GlaxoSmithKline, Research Triangle Park, USA, 3 Greenford, UK, 4 Division of Gastroenterology, University of Michigan, Ann Arbor, Michigan, USA * Dr Daniel M. Rosenberg, GlaxoSmithKline, Greenford Road, Greenford, Middlesex UB6 0HE, UK. E-mail: dr74409@gsk.com

> Hepatitis B is a serious disease that is endemic in many parts of the world. A significant proportion of patients with chronic hepatitis B (CHB) are infected with a variant form of the hepatitis B virus (HBV) which decreases or abolishes the production of hepatitis B e-antigen (HBeAg). The purpose of this literature review is to describe the epidemiology of HBeAg-negative CHB (e-CHB) worldwide. A literature search was conducted to identify studies pertaining to e-CHB and underlying variants (precore and core promoter). Fifty studies were included in our analysis. The median prevalence of e-CHB among patients with chronic HBV infection was 33% in the Mediterranean, 15% in Asia Pacific, and 14% in the USA and Northern Europe. The pre core stop codon variant was detected in a median of 60% (range 0–100%) of HBeAg-negative patients overall, 92% in the Mediterranean, 50% in Asia Pacific and 24% in the USA and Northern Europe. There were very few data on the prevalence of core promoter variants outside Asia where the median prevalence among HBeAg-negative patients was 77%. This literature review revealed that e-CHB is more common than previously suspected and that it is present worldwide with marked variations in the prevalence of associated HBV variants across different geographical regions. Additional research using population based samples of adequate size based on a consensus definition of e-CHB and using standardized HBV DNA assays is needed to better estimate the true prevalence of e-CHB and its associated HBV variants.

... 有报道亚洲CHB患者中,HBeAg阴性的乙型肝炎在日本占19%,在中国大陆占21%40%,中国台湾为20%,香港为11%,印度为16%^[1] ...

2011

0.0

... 有研究发现,DcR3具有潜在的作为炎性疾病的多功能免疫调节分子和标志物的临床应用价值,其在许多炎症性疾病中表达上调,如细菌性感染、肾炎、类风湿性关节炎、急性溃疡性结肠炎、阑尾炎等^[2,3] ...

2011

4.576

0.0

. 2011, 81(7):838-847 DOI:10.1016/j.bcp.2011.01.011

Decoy receptor 3: a pleiotropic immunomodulator and biomarker for inflammatory diseases,autoimmune diseases and cancer

Abstract Recently, several decoy molecules belonging to tumor necrosis factor receptor superfamily (TNFRSF) have been identified, including decoy receptor 1 (DcR1), decoy receptor 2 (DcR2), and decoy receptor 3 (DcR3). One of the tumor necrosis factor superfamily (TNFSF) members, TNF-related apoptosis-inducing ligand (TRAIL), binds to DcR1 and DcR2, which are membranous receptors with a truncated cytoplasmic domain, thus unable to transduce TRAIL-mediated signaling. In contrast to DcR1 and DcR2, DcR3 is a soluble receptor capable of neutralizing the biological effects of three other TNFSF members: Fas ligand (FasL/TNFSF6/CD95L), LIGHT (TNFSF14) and TNF-like molecule 1A (TL1A/TNFSF15). Since FasL is a potent apoptosis- and inflammation-inducing factor, LIGHT is involved in apoptosis and inflammation, and TL1A is a T cell costimulator and is involved in gut inflammation, DcR3 can be defined as an immunomodulator on the basis of its neutralizing effects on FasL, LIGHT, and TL1A. Initial studies demonstrated that DcR3 expression is elevated in tumors cells; however, later work showed that DcR3 expression is also upregulated in inflammatory diseases, where serum DcR3 levels correlate with disease progression. In addition to its neutralizing effect, DcR3 also acts as an effector molecule to modulate cell function via ‘non-decoy’ activities. This review focuses on the immunomodulatory effects of DcR3 via ‘decoy’ and ‘non-decoy’ functions, and discusses the potential of DcR3 as a biomarker to predict cancer invasion and inflammation progression. We also discuss the possible utility of recombinant DcR3 as a therapeutic agent to control autoimmune diseases, as well as the potential to attenuate tumor progression by inhibiting DcR3 expression. Graphical abstract

2004

0.0

... 有研究者利用转基因技术研究发现,DcR3能影响巨噬细胞的定向分化,从而抑制机体免疫应答,发挥抗炎作用^[4,5] ...

2008

9.06

0.0

... 有研究者利用转基因技术研究发现,DcR3能影响巨噬细胞的定向分化,从而抑制机体免疫应答,发挥抗炎作用^[4,5] ...

2005

3.771

0.0

. 2005, 115(3):null-null

Increased expression of soluble decoy receptor 3 in acutely inflamed intestinal epithelia

... Kim等^[6]研究发现,急性阑尾炎患者的阑尾上皮细胞中高表达DcR3,体外利用脂多糖(LPS)刺激肠上皮细胞系SW480、SW620和HT29,发现DcR3释放明显增加 ...

2010

3.771

0.0

. 2010, 137(2):null-null

Siakavellas SI,et al. High intestinal and systemic levels of decoy receptor 3 (DcR3) and its ligand TL1A in active ulcerative colitis

... 在急性溃疡性结肠炎发病过程中,DcR3的表达与炎症程度密切相关,在炎症高发期患者外周血DcR3呈现高水平表达,经有效治疗后,外周血DcR3含量明显降低^[7] ...

2010

0.0

... Yang等^[8]研究表明,肝癌患者血清中DcR3明显升高,且血清中含量与肝硬化、肿瘤转移和复发相关,与肝癌组织中DcR3表达和分布高度相关,提示DcR3在原发性肝癌的发病、转移和复发过程中发挥了重要作用 ...