摘要
目的:探讨乙型肝炎病毒(hepatitis B virus,HBV)S基因的反义锁核酸(LNA)对乙型肝炎转基因小鼠HBV复制和表达的影响.方法:将30只HBV转基因小鼠随机分为5组,每组6只.第1组为5%GLU液对照组,第2组为空脂质体对照组,第3组为单LNA组,第4组为S-ASODN脂质体组,第5组LNA脂质体组.反义LNA经尾静脉注入小鼠体内,采用ELISA法检测血清HBsAg;PCR定量检测血清HBVDNA含量;免疫组织化学法检测肝细胞HBsAg的表达;自动生化分析仪检测ALB、ALT、BUN、CR、ApoA1、ApoB等指标;小鼠肝脏、肾脏做常规病理切片HE染色,观察反义LNA对小鼠脏器的影响.结果:注射反义LNA1d、3d、7d、14d后,LNA-脂质体组对血清HBsAg的表达抑制率分别为41.7%、52.8%、57.8%及30.5%.对HBVDNA的抑制率分别为18.5%、36.1%、52.9%和32.7%,与对照组比较均有显著性差异(P<0.05);全自动生化分析仪检测血清中ALB、ALT、BUN、CR、ApoA1、ApoB等指标,各组结果与对照组比较均无显著性差异(P>0.05);小鼠肝细胞HBsAg的表达显著低于对照组.HE染色显示小鼠肝肾功能及组织学未见异常.结论:HBVS基因反义LNA对乙型肝炎病毒转基因鼠HBV复制和表达有显著抑制作用.
AIM：To investigate the inhibitory effects of hepatitis B virus（HBV）S gene-specific antisense locked nucleic acid（LNA）on HBV replication and expression. METHODS：Thirty HBV transgenic mice were randomly divided into five groups （n=6）：glucose（5%GLU solution）control group,empty liposome control group,LNA group,S-ASODN-liposome group and LNAliposome group.Antisense LNA was injected into mice via the tail vein.Serum HBsAg was quantified by ELISA.Serum HBV DNA was quantified by PCR.The expression of HBsAg in the liver was detected by immunohistochemistry.Serum ALB,ALT,BUN,CR, ApoA1 and ApoB were measured using an automatic biochemical analyzer.The effects of antisense LNA on mouse organs were investigated by HE staining of mouse liver and kidney sections. RESULTS：On days 1,3,7 and 14 after LNA injection,serum HBsAg levels in the LNAliposome group were reduced by 41.7%,52.8%, 57.8%and 30.5%,respectively,while serum HBV DNA expression levels were decreased by 18.5%,36.1%,52.9%and 32.7%,respectively. These values were significantly higher than those in the control groups（all P〈0.05）.No significant differences were noted in serum ALB, ALT,BUN,CR,ApoA1 and ApoB between the experiment group and the control groups（all P〉0.05）.The expression level of HBsAg in the liver in the LNA-liposome group was significantly lower than those in the control groups.No significant histological abnormalities were found in the liver in all groups. CONCLUSION：HBV S gene-specific antisense LNA can significantly inhibit the replication and expression of HBV.
出处
《世界华人消化杂志》
CAS
北大核心
2009年第23期2338-2345,共8页
World Chinese Journal of Digestology
基金
广州市科技攻关基金资助项目
No.2002Z3-E4081~~
关键词
脂质体
反义锁核酸
乙型肝炎病毒
小鼠
转基因
基因治疗
Cationic liposomes Antisense locked nucleic acid Hepatitis B virus Mouse Transgene Gene therapy
