HBcAg在肝细胞中的表达及其与抗病毒疗效的关系

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

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。

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 gradesⅠ,Ⅱ,Ⅲ,andⅣexpression.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 gradeⅠ,Ⅱ,Ⅲ,andⅣexpression(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 gradeⅠ,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.