胸腺因子D抗乙型肝炎病毒的体外实验  

An in Vitro Study of Anti-Hepatitis B Virus Effect of Thymic Factor D

作　　者：李丹 刘小朋 施水兰 陈少华 邓永岳 李东良 杨才生 陈紫榕 

机构地区：[1]空军福州肝病研究所,福州350002

出　　处：《海峡药学》2004年第1期73-75,共3页Strait Pharmaceutical Journal

摘　　要：目的 　探讨胸腺因子 D( TFD)在体外对乙型肝炎病毒的抑制作用及其作用的分子机制。方法 　以 He PG2 .2 .15细胞株为模型 ,应用酶联免疫吸附法 ( EL ISA)观察 TFD在不同浓度、不同作用时间对 He PG2 .2 .15细胞培养上清中乙型肝炎病毒表面抗原 ( HBs Ag)和 e抗原 ( HBe Ag)水平的影响 ,计算药物对 HBs Ag和 HBe Ag的抑制率。采用四甲基噻唑兰 ( MTT)比色法测定细胞毒性作用 ,并计算细胞的存活率和治疗指数( TI)。采用 DNA斑点杂交技术检测 Hep G2 .2 .2 .15细胞培养上清中的 HBV DNA含量的变化 ,初步探讨药物作用的分子机制。同时以 IFNα- 2 b为对照 ,综合评价 TFD的体外抗 HBV效果。结果 　 TFD对 HBs Ag、HBe Ag的 5 0 %抑制浓度分别为 48.4mg· m L- 1和 75 1.7m g·m L- 1 ,治疗指数 ( TI)分别为 2 4.2和 1.6。在用药的第 9天 ,5 0 0 mg· m L - 1的 TFD对 HBs Ag和 HBe Ag的抑制率分别 80 .63 %和 5 1.2 6%。 DNA斑点杂交结果显示 ,TFD对细胞中游离 DNA无明显抑制作用。结论 　TFD及 IFNα- 2 b在体外具有较显著的抗 HBV作用 ,TFD对 HBs Ag的抑制作用较IFNα- 2 b更为明显 ,但对 HBe Ag的抑制不如 IFNα- 2 b。TFD对 HBV的抑制可能不是通过抑制 HBV DNA的复制来实现的。OBJECTIVE To investigate the inhibitory ablity on hepatitis B virus(HBV) and mechanism of thymic factor D (TFD) in vitro.METHODS ELISA was used to observe the effect of various concentrations of TFD on the hepatitis B virus surface antigen (HBsAg) and hepatitis B virus e antigen (HBeAg)in the HePG2.2.15 cell line, and the inhibitory rate was calculated. MTT method was used to evaluate the cytotoxicity. The cell viability and therapeutic index (TI) was calculated. DNA dot blot hybridization technique was also used to detect the amounts of HBV DNA in the TFD treated and untreated cells. At the same time, we took interferon alpha-2b (IFN α-2b )as control to evaluate the anti-HBV activity of TFD in vitro.RESULTS The results showed that ID50 (the drug concentration that inhibits HBsAg or HBeAg secretion by 50%) was 48.4mg·mL -1 and 751.7mg·mL -1 respectively. The theraputic index (TI) was 24.2 and 1.6.The inhibitory rate of 500mg·mL -1 TFD on HBsAg and HBeAg was 80.63% and 51.26% respectively when 9 d. The results of the dot blotting showed TFD had no remarkable inhibitory effect on the free HBV DNA synthesis in the cell.CONCLUSION TFD and IFN α-2b showed remarkably inhibitory effect on HBV viral protei nexpression. the inhibitory effcet on HBsAg of TFD is superior to IFN α-2b in vitro,but the inhibitory effect on HBeAg of TFD is underabundant to IFN α-2b . The results of the dot blotting suggested that TFD probably has not affected HBV DNA synthesis.

关 键 词：胸腺因子D 干扰素 乙型肝炎病毒 HEPG2.2.15细胞株 体外实验 抗病毒活性 

分 类 号：R512.62[医药卫生—内科学] R373.21[医药卫生—临床医学]