甲泼尼龙和他克莫司在体外对HepG2．2．15细胞中乙型肝炎病毒cccDNA合成的影响  

Effects of methylprednisolone and tacrolimus on cccDNA replication of hepatitis B virus in HepG 2. 2. 15 cells

作　　者：沈中阳[1] 郑卫平[1] 宋红丽[1] 王建[1] 

机构地区：[1]天津市第一中心医院器官移植中心,300192

出　　处：《中华器官移植杂志》2009年第12期745-748,共4页Chinese Journal of Organ Transplantation

基　　金：天津市卫生局科研资助项目（05KYZ24）

摘　　要：目的探讨甲泼尼龙（MP）和他克莫司（Tac）在体外对乙型肝炎病毒（HBV）cccDNA合成的影响。方法采用HepG2．2．15细胞（HBV永久转染人肝癌细胞系）为体外实验模型，经四甲基偶氮唑盐（MTT）试验确定MP的安全浓度在0-250ng／ml，Tac的安全浓度在0-500ng／ml。将不同安全浓度的MP（0、10、50、100及250ng／ml）和Tac（0、50、100及500ng／ml）分别作用于该细胞系，72h后收集细胞培养上清液及细胞，采用实时定量聚合酶链反应检测上清液中HBV DNA及细胞内HBV cccDNA水平。结果与不含MP者比较，当MP浓度分别为10、50、100及250μg／L时，HBV DNA和HBV cccDNA水平明显降低，分别为（5．7823±0.1861）、（5．1337±0．1364）、（4．7865±0．0398）及（4．1468±0．1016）log10拷贝／ml（P〈0．015，P〈0．01）；HBV cccDNA水平分别为（5．8975±0．0296）、（5．7471±0．0524）、（5．1162±0．0039）及（4．8272±0．1394）log10拷贝／ml（P〈0．05，P〈0．0t），该抑制作用呈剂量依赖性。不同Tac浓度组的HBV DNA和HBV cccDNA水平虽然低于不含Tac者，但差异均无统计学意义（P〉0．05）。结论在体外．MP能抑制HepG2．2．15细胞中HBV DNA的表达，同时可以减少细胞内HBV cccDNA的合成，该作用呈剂量依赖性；Tac无此作用。Objective To investigate the effect of methylprednisolone （MP） and tacrolimus （Tac） on hepatitis B virus （HBV） cccDNA in vitro. Methods The HepG 2.2. 15 cells were the line of HBV genome permanently transfected to human liver cancer cells. MTT was used to determine the nontoxic concentrations of MP and Tac. HepG 2.2. 15 cells were treated with safe concentrations of MP and Tac for 72 h. Intracellular HBV cccDNA and supernatant HBV DNA were analyzed by real time polymerase chain reaction （RT-PCR）. Results MTT revealed that the nontoxic concentrations of MP and Tac were 0-250 ng/ml and 0-500 ng/ml, Respectively. After treatment with MP at the concentrations of 10.50, 100 and 250 ng/ml, the HBV DNA replication levels were 5. 7823±0. 1861, 5. 1337 ＋ 0. 1364,4. 7865 ±0. 0398 and 4. 1468 ＋ 0. 1016 log10 copy/ml, respectively （P〈0. 05 and P〈0.01 ）,and the cccDNA replication levels were 5. 8975 ± 0. 0296, 5. 7471±0. 0524, 5. 1162 ±0. 0039 and 4. 8272 ±0. 1394 log10 copy/ml, respectively （P〈0. 115 and P〈0. 01 ）, which were significantly lower than in control group （6. 11859 ± 0. 2014 and 6. 8164±0. 0753 log10 copy/ml）. MP suppressed the replication of HBV DNA and cccDNA. However, Tac at different nontoxic concentrations showed no significantly inhibitory effects on the levels of HBV DNA and cccDNA. Conclusion MP dose- dependently inhibits the HBV replication in vitro. Reducing the synthesis of cccDNA may be involved in the mechanism of the inhibitory activity of MP on HBV replication. Tac does not exercise the similar effects.

关 键 词：甲泼尼龙 他克莫司 肝炎病毒 乙型 基因 合成 

分 类 号：R735.7[医药卫生—肿瘤] R512.62[医药卫生—临床医学]