半乳糖受体介导的拉米夫定肝细胞导向作用的体内外研究

ANTI-HBV EFFECT OF GALACTOSIDES MODIFIED LAMIVUDINE TARGETED TO LIVER

探讨修饰后拉米夫定 (lamivudine,LA)对乙型肝炎病毒 (HBV)的抑制作用及肝细胞的靶向性。以半乳糖苷为载体 ,构建十六酸拉米夫定酯固体脂质纳米粒(LAP GSLN) ,将其导入 2 2 15细胞 ,用免疫荧光法及ELISA观察 2 2 15细胞内HBeAg及用免疫定量PCR检测细胞培养上清中HBVDNA ;将LAP GSLN经尾静脉注入小鼠体内 ,用RP HPLC测定LA在血清、肝、肾、肺及脾中的分布。结果显示 ,LA及LAP GSLN对 2 2 15细胞HBeAg的表达和对HBV DNA的复制均有抑制作用 ;LAP GSLN组肝脏的含药量为LA组的3 3倍。表明LAP GSLN能有效地抑制HBeAg表达和HBVDNA的复制 。

The aim of the present study was to investigate the inhibitory effect of galactosides modified lamivudine (LA) on hepatitis B virus (HBV) and examine the liver targeting ability of lamivudine modified by galactosides in vitro and in vivo (mice). Lamivudine nanoparicles modified by galactosides (LAP GSLN) were prepared and delivered into 2.2.15 cells. After 10 days, hepatitis virus B e antigen (HBeAg) expression in 2.2.15 cells was detected by ELISA, and immune fluorescence levels of HBV DNA in the medium were examined by quantitative polymerase chain reaction (PCR). The cytotoxicity of LAP GSLN on 2.2.15 cells was observed as well. In the in vivo experiment, ten male mice were randomly divided into 2 groups: lap GSLN group (i.v.injection of LAP GSLN) and LA group (i.v.injection of LA). Lamivudine levels in serum, hepatic, renal, pulmonary, and splenic tissues were detected by reversed phase high performance liquid chromatography (RP HPLC). On the 6th day, the expression of HBeAg was found inhibited by LPA GSLN. HBV DNA replication was also inhibited by LAP GSLN on the 4th day. Hepatic LAP GSLN concentrations in LAP GSLN group were 3.3 fold higher that of the LA group. The above results suggested galactosides modified lamivudine could effectively inhibit the antigen expression and DNA replication of HBV, and it showed a high liver targeting ability in vivo .