反义RNA寡核苷酸及其与脂质体的复合物对于乙型肝炎病毒的抑制（英文）  

Inhibition of Hepatitis B Virus in Mice by Cationic Liposome/RNA Oligonucleotide Complex

作　　者：昝琰璐[1,2] 田波[1] 

机构地区：[1]中国科学院微生物研究所,中国科学院病原微生物与免疫学重点实验室,北京100101 [2]中国科学院大学,北京100101

出　　处：《生物化学与生物物理进展》2014年第4期362-370,共9页Progress In Biochemistry and Biophysics

基　　金：supported by grants from The National Basic ResearchProgram of China(2011CB504703,2010CB530102);The NationalNatural Science Foundation of China(81021003)~~

摘　　要：我们设计合成了特异性靶向乙型肝炎病毒(HBV)mRNA的反义RNA寡核苷酸P-2987、X-60和X-519.在瞬时转染pHBV1.3质粒(含有1.3拷贝的HBV基因组)的HepG2细胞和整合了HBV基因组的HepG2.2.15细胞中,转染2μmol/L的反义RNA寡核苷酸,ELISA和实时定量PCR结果表明,这3条寡核苷酸可以明显抑制HBV的复制和抗原表达.在HBV转基因鼠中,尾静脉注射反义RNA寡核苷酸,结果表明,肝脏中HBV的复制得到了抑制,但是血清中抗原含量和HBV DNA拷贝数没有明显变化.反义RNA寡核苷酸X-519与脂质体的复合物可以增强其对于HBV在肝脏中复制的抑制作用.在通过高压尾静脉注射pHBV1.3质粒建立的HBV急性感染模型中,反义RNA寡核苷酸X-519可以显著地抑制HBV在肝脏中的复制以及降低血清中病毒抗原水平和DNA拷贝数.上述实验结果说明,X-519及其与脂质体的复合物对于HBV的复制和抗原表达起到明显的抑制作用,可能作为一种潜在的针对HBV的基因治疗药物.We investigated the inhibitory effect of modified antisense RNA oligonucleotides and cationic liposome-RNA complexes on the repression of hepatitis B virus （HBV） replication and expression. ELISA and quantitative Real-time PCR analysis showed that HBV replication and antigens expression both in pHBV1.3 transduced HepG2 and HepG2.2.15 cells were reduced after treatment with antisense RNA oligonucleotides P-2987, X-60 and X-519. Subsequently, the antisense RNA oligonucleotides and control RNA oligonucleotides were injected via the tail vein into I-IBV transgenic mice or hydrodynamically injected mice. In the HBV transgenic mice, with the treatment of X-519, HBV pregenomic RNA in the liver decreased by 81%. Cationic liposome further increased the inhibition effectory to 91%. But no significant differences were observed for HBV antigens and HBV DNA copy number in the sera. In acute HBV infection mouse model by hydrodynamic injection, ELISA and quantitative real-time PCR analysis showed that X-519 significantly repressed HBV replication, as measured by HBV pregenomic RNA, antigens expression, and presence of HBV DNA in the sera. Taken together, the synthesized antisense RNA oligonucleotide X-519 repressed HBV replication and antigens expression in vitro and in vivo.

关 键 词：乙型肝炎病毒 反义RNA寡核苷酸 HBV转基因鼠 高压尾静脉注射 反义RNA寡核苷酸与脂质体复合物 

分 类 号：Q78[生物学—分子生物学]