地塞米松、环磷酰胺和他克莫司三种免疫抑制剂影响HBV复制的体外研究

肝脏 ›› 2018, Vol. 23 ›› Issue (5): 391-394.

地塞米松、环磷酰胺和他克莫司三种免疫抑制剂影响HBV复制的体外研究

黄顺梅, 王俊忠, 张小勇, 李宝林, 宋志韬, 朱珍妮, 朱彬, 王宝菊, 陆蒙吉, 杨东亮

430022 武汉华中科技大学同济医学院附属协和医院感染科(黄顺梅,王俊忠,宋志韬,朱珍妮,朱彬,王宝菊,杨东亮);德国杜伊斯堡埃森大学病毒研究所(张小勇,李宝林,陆蒙吉)

收稿日期:2018-02-04 出版日期:2018-05-31 发布日期:2020-04-28
通讯作者: 王宝菊,Email:bjwang73@yahoo.com
基金资助:
国家艾滋病和病毒性肝炎等重大传染病防治专项(No.2008ZX10002011)

Effect of three immunosupressive drugs dexamethasone, cyclophosphamide and tacrolimus on HBV replication in vitro

HUANG Shun-mei, WANG Jun-zhong, ZHANG Xiao-yong, LI Bao-lin, SONG Zhi-tao, ZHU Zhen-ni, ZHU Bin, WANG Bao-ju, LU Meng-ji, YANG Dong-liang

Department of Infectious Disease, Union Hospital of Tongji Medical college, Huazhong University of Science and Technology, Wuhan, China; and Institute of Virology, University Hospital Essen, Germany

Received:2018-02-04 Online:2018-05-31 Published:2020-04-28
Contact: WANG Bao-ju, Email: bjwang73@yahoo.com

摘要/Abstract

摘要： 目的探讨地塞米松(DEX)、环磷酰胺(CYP)和他克莫司(FK506)在体外对HBV蛋白合成、DNA复制的影响。方法将不同浓度的DEX、CYP和FK506作用于HepG2.2.15细胞系,通过MTT实验确定药物安全浓度范围,通过检测细胞培养上清液中HBsAg和HBeAg水平,以及细胞内HBV DNA水平来评价HBV的表达和复制情况。结果 DEX、CYP和FK506的药物安全浓度范围分别为0～500、0～1000和0～10 μg/mL。与培养基对照处理相比,FK506处理后HBsAg、HBeAg分泌及HBV DNA复制变化差异无统计学意义;CYP处理增加HBsAg分泌,与对照相比差异有统计学意义(P<0.05);但HBeAg分泌及HBV DNA复制变化差异无统计学意义;DEX作用减少HBsAg和HBeAg的分泌,与对照相比差异有统计学意义(P<0.05);但是细胞内HBV DNA复制水平差异无统计学意义(P>0.05)。结论在药物安全浓度范围内,FK506对HepG2.2.15细胞HBV DNA复制无直接抑制或促进作用。CYP处理后增加HepG2.2.15细胞HBsAg表达,但HBeAg表达及HBV DNA复制变化差异无促进作用;DEX处理抑制HepG2.2.15细胞HBsAg和HBeAg表达,但对 HBV DNA复制无抑制作用。

关键词: 免疫抑制剂, 乙型肝炎病毒, HepG2.2.15细胞

Abstract: Objective To investigate the effect of dexamethasone (DEX), cyclophosphamide (CYP) and tacrolimus (FK506) on DNA replication and protein synthesis of hepatitis B virus (HBV) in vitro. Methods HepG2.2.15 cell line was treated with different concentrations of DEX, CYP or FK506, and safe concentrations of the three immunosupressive drugs were identified using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Levels of hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) in the cell culture supernatant were detected with electrochemiluminescence immunoassay. The intracellular HBV DNA level was evaluated with Southern blot. HBV expression and replication were assessed according to HBsAg, HBeAg and HBV DNA level. Results The safe concentration range of DEX, CYP and FK506 was 0～500 μg/mL, 0～1000 μg/mL and 0～10 μg/mL, respectively. Compared with control group, FK506 group showed no significant difference in the levels of HBsAg, HBeAg and HBV DNA. CYP group showed higher levels of HBsAg (P<0.05), while no significant difference in the levels of HBeAg and HBV DNA (P>0.05). DEX group showed lower levels of HBsAg and HBeAg (P<0.05), while no significant difference in HBV DNA (P>0.05). Conclusion In the safe concentration range, FK506 has no direct inhibitory or promoting effect on HBV DNA replication. CYP treatment could increase HBsAg expression, but have no effect on HBeAg expression and HBV DNA replication. DEX treatment could inhibit the expression of HBsAg or HBeAg, but has no inhibitory effect on HBV DNA replication.

Key words: Immunosupressive drugs, Hepatitis B virus, HepG2.2.15 cell

黄顺梅, 王俊忠, 张小勇, 李宝林, 宋志韬, 朱珍妮, 朱彬, 王宝菊, 陆蒙吉, 杨东亮. 地塞米松、环磷酰胺和他克莫司三种免疫抑制剂影响HBV复制的体外研究[J]. 肝脏, 2018, 23(5): 391-394.

HUANG Shun-mei, WANG Jun-zhong, ZHANG Xiao-yong, LI Bao-lin, SONG Zhi-tao, ZHU Zhen-ni, ZHU Bin, WANG Bao-ju, LU Meng-ji, YANG Dong-liang. Effect of three immunosupressive drugs dexamethasone, cyclophosphamide and tacrolimus on HBV replication in vitro[J]. Chinese Hepatolgy, 2018, 23(5): 391-394.

[1]	黄珊珊, 赵静雅, 陈洁, 杨扬. 前列腺素E2促进乙型肝炎病毒复制的机制研究[J]. 肝脏, 2020, 25(3): 277-281.
[2]	龙云铸, 贺萧瑾, 李丹, 周娟, 周青, 谭英征. HBx介导SOCS-1基因甲基化致肝细胞癌发生的机制[J]. 肝脏, 2020, 25(2): 202-205.
[3]	王贤, 林涛发, 詹志瑜, 王少扬. 不同免疫状态慢性HBV感染患者的T淋巴细胞及NKG2D表达差异研究[J]. 肝脏, 2020, 25(2): 131-134.
[4]	王欣茹, 肖丽, 杨莉莉, 张会, 汤小霞, 孙鸿展, 咸建春. 不同年龄孕妇HBV感染现状的血清流行病学分析[J]. 肝脏, 2020, 25(1): 76-77.
[5]	华燕美. 两种乙型肝炎病毒核酸定量检测试剂的检验重复性与一致性比较[J]. 肝脏, 2019, 24(6): 692-694.
[6]	钟慧筠, 殷思纯. HBV相关慢加急性肝衰竭器官衰竭危险因素及短期预后分析[J]. 肝脏, 2019, 24(6): 658-661.
[7]	曹香芸, 管艳萍, 李倩, 周晓敏, 刘高伟. 替诺福韦、替比夫定分别联合双重免疫方案对HBV母婴传播阻断效果的比较[J]. 肝脏, 2019, 24(5): 503-506.
[8]	苏金良, 陈慧, 朱莉, 陆敏嘉, 李明. HBV相关慢加急性肝衰竭前期外周血中Toll样受体3、4的表达[J]. 肝脏, 2019, 24(5): 499-502.
[9]	薛扬, 郭宏强. 血清miRNA-130a与HBV相关慢加急性(亚急性)肝衰竭患者预后的相关性分析[J]. 肝脏, 2019, 24(2): 158-160.
[10]	谢玲, 程丰, 孙文锦, 龚国富. 血清同型半胱氨酸联合降钙素原检测对HBV相关慢加急性肝衰竭患者短期预后的临床价值[J]. 肝脏, 2019, 24(12): 1402-1405.
[11]	吴燕玉, 陈红敏, 徐秋霞. 乙型肝炎病毒感染对妊娠期肝内胆汁淤积症患者妊娠结局的影响[J]. 肝脏, 2019, 24(11): 1303-1306.
[12]	武文辉, 崔立春, 党升强, 王剑. HBV相关性肝癌患者血清B7-H3、IL-21水平及MRI影像学变化与患者病情严重程度的关系[J]. 肝脏, 2019, 24(11): 1251-1254.
[13]	顾静, 陈祖涛, 孙蔚, 甘建和. 乙型肝炎相关慢加急性肝衰竭患者血清碱性磷酸酶的变化及意义[J]. 肝脏, 2019, 24(11): 1234-1236.
[14]	王艳巧, 陈传杰, 张光文. 乙型肝炎病毒相关原发性肝癌的危险因素分析[J]. 肝脏, 2019, 24(10): 1180-1181.
[15]	王兆勋, 周渐, 施文娟, 张妍, 万红. 98例HBV相关慢加急性肝衰竭患者的临床特征与分析[J]. 肝脏, 2019, 24(10): 1176-1177.