双靶区反义RNA对HBV转基因鼠抗病毒疗效的研究

发布时间：2018-02-11 07:06

本文关键词： 乙型肝炎病毒反义RNA 转基因小鼠　出处：《兰州大学》2006年硕士论文　论文类型：学位论文

【摘要】：目的探讨针对乙型肝炎病毒(HBV)x、p双靶区反义RNA对乙型肝炎病毒转基因小鼠HBV复制和表达的影响。方法将表达HBV X、P双靶区正、反义RNA的重组载体质粒与脂质体混合,经尾静脉注入小鼠体内,用酶联免疫法检测其血清HBsAg:用荧光聚合酶链反应定量方法检测血清HBV DNA含量;用免疫组化法检测心脏、肝脏、脾脏、肺脏、肾脏细胞HBsAg、HBcAg的表达;小鼠心脏、肝脏、脾脏、肺脏、肾脏常规病理切片,观察反义RNA对小鼠脏器的影响。结果与给药前相比,小鼠血清HbsAg表达,在PLXSN-asX组和PLXSN-asXP组均于第四周后出现明显下降,最高抑制率分别达24%、27%;其余组未出现明显变化。与给药前相比,小鼠血清HBV DNA的复制,在PLXSN-asX组和PLXSN-asP组分别于第2周、第8周达到抑制高峰,抑制率均为58%;在PLXSN-asXP组于第1周、第8周出现两次抑制高峰,抑制率分别为66%、77%;其余组未出现明显变化。8周后,PLXSN-asX组、PLXSN-asP组和PLXSN-asXP组小鼠肝细胞HBsAg、HBcAg的表达显著低于其它组。所有小鼠心脏、脾脏、肺脏未见HBsAg及HBcAg的表达;肾脏组织细胞中未见HBsAg的表达,但除PLXSN-asXP组外,其余组均有小鼠肾曲小管上皮细胞表达HBcAg。小鼠脏器组织学观察未见异常。结论 HBV X、P双靶区反义RNA对乙型肝炎病毒转基因鼠有显著抗HBV作用,且在作用效率和作用时间上优于单靶区反义RNA。
[Abstract]:Objective to investigate the effect of antisense RNA targeting HBV RNA on the replication and expression of HBV in HBV transgenic mice. Methods Recombinant vector plasmid expressing sense and antisense RNA of HBV XNP was mixed with liposome and injected into mice via tail vein. Serum HBsAg was detected by enzyme linked immunosorbent assay (Elisa) and serum HBV DNA was detected by fluorescence polymerase chain reaction (FPCR). The expression of HBcAg in heart, liver, spleen, lung and kidney was detected by immunohistochemical method, and the effects of antisense RNA on viscera were observed by routine pathological sections of heart, liver, spleen, lung and kidney of mice. Results compared with before administration, the expression of HbsAg in serum of mice was significantly decreased in PLXSN-asX group and PLXSN-asXP group after the fourth week, and the highest inhibitory rate was 240.27%, while no significant change was found in the other groups. Compared with before administration, the replication of serum HBV DNA was observed in mice. In PLXSN-asX group and PLXSN-asP group, the inhibition peak was reached at the 2nd week and the 8th week, respectively, and the inhibition rate was 58% in the PLXSN-asXP group, and the inhibition peak occurred twice in the PLXSN-asXP group at the first week and the eighth week in the PLXSN-asXP group. The inhibitory rate of HBcAg was 66 / 777.The expression of HBcAg in hepatocytes of mice in PLXSN-asP group and PLXSN-asXP group was significantly lower than that in other groups. No expression of HBsAg and HBcAg was found in heart, spleen and lung of all mice. The expression of HBsAg was not found in renal tissue cells, but the expression of HBcAg in the epithelial cells of renal tubules was found in all the other groups except the PLXSN-asXP group. There was no abnormal expression of HBcAg in the viscera of the mice with the exception of the PLXSN-asXP group. Conclusion the antisense RNA with double target region of HBV XNP has a significant anti-HBV effect on HBV transgenic mice, and is superior to the antisense RNA of single target region in the effect efficiency and time.
【学位授予单位】：兰州大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R346

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