HBcAg在肝细胞中的表达及其与抗病毒疗效的关系

付喜花; 黄璇; 沈国俊; 娄海波; 毛玉巧

doi:10.3969/j.issn.1001-5256.2021.06.017

HBcAg在肝细胞中的表达及其与抗病毒疗效的关系

DOI: 10.3969/j.issn.1001-5256.2021.06.017

1.
广州市番禺区中心医院感染科，广州 511400
2.
南方医科大学南方医院感染科，广州 510515
3.
九江市第三人民医院肝病防治专科，江西九江 332000

基金项目:

广州市科技计划项目 (201904010065)

利益冲突声明：本研究不存在研究者、伦理委员会成员、受试者监护人以及与公开研究成果有关的利益冲突。

作者贡献声明：付喜花负责课题设计，资料分析，撰写论文; 黄璇、娄海波、毛玉巧参与收集数据，资料分析; 沈国俊负责指导修改文章并最后定稿。

详细信息

作者简介:
付喜花(1980—)，女，博士，副主任医师，主要从事病毒性肝炎的临床研究

通信作者:
付喜花，nsndys@163.com

中图分类号: R512.62
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出版历程
- 收稿日期: 2020-11-05
- 录用日期: 2020-12-22
- 出版日期: 2021-06-20

Expression of HBcAg in hepatocytes and its association with the efficacy of antiviral therapy

1.
Department of Infectious Diseases, Panyu Central Hospital, Guangzhou 511400, China
2.
Department of Infectious Diseases, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China
3.
Department of Prevention and Treatment of Liver Diseases, Jiujiang Third People's Hospital, Jiujiang, Jiangxi 332000, China

摘要

摘要: 目的研究HBcAg在肝细胞中的表达对血清抗-HBc水平和核苷类药物抗病毒治疗后HBeAg血清学转换的影响。方法收集南方医院和番禺区中心医院2015年1月—2018年6月101例接受核苷类药物抗病毒治疗的慢性乙型肝炎患者的血清样本和肝组织石蜡切片，酶联免疫吸附测定血清中的抗-HBc定量，免疫组化检测肝内HBcAg表达。通过GEO数据库(GSE96851)分析获得HBcAg阳性肝炎活动患者肝内差异基因表达。计量资料两组间比较采用两独立样本t检验, 多组间比较采用多个独立样本非参数Kruskal-Wrallis H检验，进一步两两比较采用Dunnett法。计数资料多组间比较采用χ²检验。结果 HBcAg在肝细胞中，根据表达类型可分为无表达型、核型、浆型和核浆型，根据表达量可分为Ⅰ、Ⅱ、Ⅲ和Ⅳ级。抗病毒96周的HBeAg血清学转换在无表达型、核型、浆型和核浆型患者的比例分别为5.88%、16.67%、22.73%、24.24%(χ²=4.753，P=0.037);抗病毒96周的HBeAg血清学转换在Ⅰ、Ⅱ、Ⅲ和Ⅳ级患者的比例分别为5.88%、13.04%、27.59%、26.67%(χ²=6.580，P=0.016)。血清IgM型抗-HBc、总抗-HBc在无表达型、核型、浆型和核浆型患者中差异均有统计学意义(H值分别为9.760、21.46，P值分别为0.021＜0.001);血清IgM型抗-HBc、总抗-HBc在Ⅰ、Ⅱ、Ⅲ和Ⅳ级患者中差异均有统计学意义(H值分别为18.80、26.03，P值均＜0.001)。肝内差异基因分析显示，HBcAg阳性急性肝衰竭患者，肝内抗体相关基因表达上调。结论肝细胞内HBcAg在胞浆表达类型及表达水平与血清抗-HBc有关，其检测有利于预测核苷类药物抗病毒治疗的效果。
- 乙型肝炎，慢性 /
- 乙型肝炎核心抗原 /
- 乙型肝炎e抗原 /
- 治疗结果
Abstract: Objective To investigate the effect of the expression of HBcAg in hepatocytes on the serum level of HBcAb and seroconversion of HBeAg after antiviral therapy with nucleos(t)ide analogues (NUCs). Methods Serum samples and liver tissue paraffin sections were collected from 101 chronic hepatitis B (CHB) patients who received antiviral therapy with NUCs in Nanfang Hospital and Panyu Central Hospital from January 2015 to June 2018. ELISA was used to measure the serum level of HBcAb, and immunohistochemistry was used to measure the expression of HBcAg in the liver. The GEO database (GSE96851) was analyzed to obtain differentially expressed genes in the liver of patients with HBcAg-positive hepatitis. The two-independent-samples t test was used for comparison of continuous data between two groups; the multiple-independent-samples nonparametric Kruskal-Wallis H test was used for comparison of continuous data between multiple groups, and Dunnett method was used for further comparisons; the chi-square test was used for comparison of categorical data between groups. Results The expression pattern of HBcAg in hepatocytes was classified as absent expression, nuclear expression, cytoplasmic expression, and nuclear/cytoplasmic expression, and according to expression level, HBcAg expression was classified as grades Ⅰ, Ⅱ, Ⅲ, and Ⅳ expression. HBeAg seroconversion rates after 96 weeks of antiviral therapy were 5.88%, 16.67%, 22.73%, and 24.24%, respectively, in the patients with absent expression, nuclear expression, cytoplasmic expression, and nuclear/cytoplasmic expression (χ²=4.753, P=0.037), and HBeAg seroconversion rates after 96 weeks of antiviral therapy were 5.88%, 13.04%, 27.59%, and 26.67%, respectively, in the patients with grade Ⅰ, Ⅱ, Ⅲ, and Ⅳ expression (χ²=6.580, P=0.016). There were significant differences in the serum levels of HBcAb-IgM and total HBcAb between the patients with absent expression, nuclear expression, cytoplasmic expression, and nuclear/cytoplasmic expression of HBcAg (HBcAb-IgM: H=9.760, P=0.021; total HBcAb: H=21.46, P < 0.001), and there were also significant differences in the serum levels of HBcAb-IgM and total HBcAb between the patients with grade Ⅰ, Ⅱ, Ⅲ, and IV expression of HBcAg (HBcAb-IgM: H=18.80, P < 0.001; total HBcAb: H=26.03, P < 0.001). The analysis of differentially expressed genes in the liver showed that the expression of antibody-related genes was upregulated in the liver of patients with HBcAg-positive acute liver failure. Conclusion The expression pattern and level of HBcAg in the cytoplasm of hepatocytes are associated with serum HBcAb, and the measurement of HBcAg may help to predict the efficacy of antiviral therapy with NUCs.
- Hepatitis B, Chronic /
- Hepatitis B Core Antigens /
- Hepatitis B e Antigens /
- Treatment Outcome

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图 1 HBcAg在肝细胞的表达类型

注：箭头代表HBcAg阳性表达。

下载: 全尺寸图片幻灯片

图 2 肝内不同HBcAg表达类型、分级患者血清抗-HBc的水平

下载: 全尺寸图片幻灯片

图 3 HBcAg阳性急性肝衰竭组织(4例)与对照肝组织(10例肝移植供者肝脏组织和7例血管瘤切除肝组织)肝内差异基因表达(数据来源于GSE96851)

下载: 全尺寸图片幻灯片

表 1 CHB患者基本资料

指标	HBeAg转换(n=17)	HBeAg未转换(n=84)	统计值	P值
男/女	10/7	64/20	χ²=2.177
年龄(岁)	33.7±8.5	30.2±9.6	t=1.323	0.189
ALT (U/L)	85.6±13.3	73.0±22.1	t=2.136	0.035
AST(U/L)	51.3±20.2	45.3±22.6	t=0.963	0.338
TBil (μmol/L)	5.2±2.8	6.7±3.1	t=1.752	0.083
HBV DNA (log₁₀ IU/ml)	5.81±1.81	5.94±1.53	t=0.295	0.768
HBsAg(IU/ml)	3.76±0.65	3.93±0.45	t=1.257	0.212

下载: 导出CSV

表 2 不同HBcAg表达类型、分级的患者HBeAg血清学转换情况

HBcAg表达类型	例数	HBeAg血清学转换[例(%)]	HBcAg分级	例数	HBeAg血清学转换[例(%)]
无表达组	34	2(5.88)	Ⅰ级	34	2(5.88)
核型表达组	12	2(16.67)	Ⅱ级	23	3(13.04)
浆型表达组	22	5(22.73)	Ⅲ级	29	8(27.59)
核浆表达组	33	8(24.24)	Ⅳ级	15	4(26.67)

下载: 导出CSV

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