慢性乙型肝炎患者肝组织HBV cccDNA、总HBV DNA与血清HBV DNA的相关性及其与临床的关系  

The relationship among HBV cccDNA,HBV total DNA in liver tissue and serum HBV DNA and their clinic - pathological significances in patients with chronic hepatitis B

作　　者：饶敏[1] 陆伟[1] 张占卿[1] 张小楠[1] 曹婕[1] 

机构地区：[1]复旦大学附属公共卫生临床中心肝炎二科,上海201508

出　　处：《肝脏》2012年第6期381-384,共4页Chinese Hepatology

摘　　要：目的探讨慢性乙型肝炎患者肝组织HBV共价闭合环状DNA（cccDNA）、肝组织总HBV DNA（HBV tDNA）与血清HBV DNA之间的相关性及其与临床的关系。方法78例慢性乙型肝炎患者入选本研究。肝组织β—globin DNA、HBV cccDNA和HBV tDNA采用实时荧光定量PCR方法检测，平均每个肝细胞HBV cccDNA和HBV tDNA含量（拷贝／cell）=HBV cccDNA（实测值）／β-globin DNA（实测值）和HBV tDNA（实测值）／β-globin DNA（实测值），肝组织HBV cccDNA和HBV tDNA含量的计算单位定义为log10拷贝／10^6 cell；采用实时荧光定量PCR、ELISA法检测血清HBV DNA和HBV标志物；采用免疫组织化学方法检测肝细胞中HBsAg和HBcAg的表达。统计分析采用pearson相关分析及t检验。结果（1）肝组织HBV cccDNA与HBV tDNA定量呈正相关（r=0．696，P〈0．001）；肝组织HBV cccDNA与血清HBV DNA定量呈正相关（r=0．304，P％0．01）；肝组织HBV tDNA与血清HBV DNA定量呈正相关（r=0．341，P〈0．01）；（2）肝细胞内HBcAg定性检测阳性患者的血清HBV DNA定量明显高于阴性患者，且差异有统计学意义（P〈0．05）；肝细胞内HBcAg定性检测阳性患者与阴性患者的肝组织HBV cccDNA和HBV tDNA定量差异均无统计学意义和（P均〉0．05）；（3）肝细胞内HBsAg定性检测阳性患者的血清HBV DNA定量明显高于阴性患者，且差异有统计学意义（P〈0．05）；肝细胞内HBsAg定性检测阳性患者与阴性患者的肝组织HBV cccDNA和HBV tDNA定量差异均无统计学意义（P〉0．05）；（4）HBeAg（＋）／抗HBe（-）患者血清HBV DNA定量明显高于HBeAg（-）／抗-HBe（＋）患者，且差异有统计学意义（P〈0．05）；HBeAg（＋）／抗-HBe（-）患者肝组织HBV cccDNA和HBV tDNA定量与HBeAg（-）／抗-HBe（＋）患者比较差异均无统计学意义（均P〉0.05）；（5）肝组织HBV cccDNA、HBV tDNA以及血清HBV DNA三者与肝脏炎症活动度及纤维化�Objective To investigate the relationship among covalent closed circular DNA （cccDNA）, HBV total DNA （tDNA） and serum HBV DNA and their clinic-pathological significances in patients with chronic hepatitis B. Methods Seventy eight patients with chronic hepatitis B were enrolled in this study. The β- globin DNA, HBV cccDNA and HBV tDNA in liver tissue were detected by real-time PCR. The content of single hepatocyte HBV cccDNA and HBV tDNA （cop- ies/cell） equals to HBV cccDNA （measured value）/β-globin DNA measured value（copies）, and HBV tDNA measured value （copies）/β-globin DNA measured value（copies）. The content of intrahepatic HBV cccDNA and HBV tDNA was recorded as log10 copies/10^6 cell. Real time PCR and ELISA were performed to determine serum HBV DNA and HBV markers respec- tively. The expression of HBsAg and HBcAg in hepatocytes was detected by immunohistochemical method. SPSS 13.0 soft ware was adopted for statistical analysis. Results （1） There was a positive correlation between the hepatocyte HBV cccDNA and HBV tDNA （r = 0. 696, P〈（0. 001 ） , Similar positive correlation was found between hepatocyte HBV cccDNA and serum HBV DNA （r= 0. 304, P〈0.01 ） ,. There was a positive correlation between hepatocyte HBV tDNA and serum HBV DNA （r= 0. 341, P〈0. 01） as well. （2） Serum HBV DNA in patients with intrahepatic HBcAg（ ＋ ） was higher than that in patients with intrahepatic HBcAg（ - ）, and there was a significant difference（P〈0.05）. There was no significant differ enee between hepatocyte HBV cccDNA and HBV tDNA in patients with intrahepatic HBcAg（ ＋ ） and intrahepatic HBcAg （ - ）with P〉0.05 respectively. （3） Serum HBV DNA with intrahepatic HBsAg（ ＋ ） was higher than that in patients with intrahepatic HBsAg（ - ）, and there was a significant difference（P〈0.05）. There was a positive correlation between hepa tocyte HBV cccDNA and HBV tDNA in patients with intrahepatic HBsAg（ ＋ ） and intrahepatic HBsAg（ -

关 键 词：乙型肝炎病毒 共价闭合环状DNA 肝组织 实时定量PCR 

分 类 号：R512.62[医药卫生—内科学]