针对HBV preC/C双靶区反义锁核酸抑制HBV基因表达体内实验研究

objective To investigate the curative effects of dual-target antisense LNA targeting the preC and C regions in the genome of hepatitis B virus. methods Antisense LNA complementary to the preC and C regions in the genome of HBV was synthesized respectively. Cationic liposomes were used as drugs carrier which could transport antisense LNA to mice liver. 30 HBV transgenic mice were randomly divided into 5 equal groups: single-target preC group, single-target C group, dual-target preC/C group, blank liposomes control group, and 5%GLU control group. LNA was incubated with cationic liposomes for 60 minutes to produce the targeted Lipo-LNA mixture which was injected into the mice via caudal vein one time every day. While in the control group, each mouse received the same volume 5%GLU solution or blank liposomes in the same way. Venous blood samples were collected before, 1, 3 and 5 d after injection.HBV DNA was detected by Real Time PCR. While serum Alb, ALT, BUN and CR were measured by automatic biochemistry analyzer. 5 d later the mice were killed and RT-PCR was used to detect the expression of mRNA. results The serum HBV DNA concentrations 1, 3 and 5 d after injection were significantly lower than that before injection in the dual-target preC/C group(P <0.05). The inhibition rate was 18.52%, 36.12% and 53.72%. HBV C- mRNA in liver were significantly lower than control group in 5 d later. No significant abnormality was found with the liver function in all groups. conclusion Dual-target antisense LNA targeting the preC and C regions in the genome of HBV inhibits the replication and expression of HBV, significantly stronger than single-target antisense LNA.