【摘要】 目的 探索纳米载体介导的LNA反基因锁核酸(LNA antigene)在乙型肝炎病毒(HBV)转基因小鼠模型中对HBV的抑制效果。方法 将LNA antigene与聚乳酸-羟基乙酸共聚物结合，制备LNA antigene纳米颗粒(NP),并通过纳米颗粒跟踪分析(NTA)和Zeta电位测定其粒径和表面电位。评估其包封率、载药量及在HepG2肝癌细胞中的摄取效率。随后，分析LNA antigene-NP的体外释放特性及其对转基因小鼠HBV DNA、HBsAg和HBeAg表达的抑制效果。结果 成功制备LNA antigene-NP纳米颗粒，其粒径集中在125～225 nm之间，表面电位较未修饰的NP有所降低。LNA antigene-NP的包封率为32%,载药量为8.448 mg。在HepG2肝癌细胞中，LNA antigene-NP显示出显著的细胞摄取效率。体外释放实验显示，LNA antigene-NP在24天内表现出持续释放的特点，最终释放率接近90%。在转基因小鼠中，LNA antigene-NP对HBV DNA、HBsAg和HBeAg的抑制效果呈剂量依赖性，5 kg/mg剂量组在第7天的抑制率分别达到约40%、60%和60%。结论 LNA antigene-NP纳米颗粒能够有效抑制HBV的复制和抗原表达，为进一步开发基于LNA的抗HBV治疗提供了实验基础。更多还原

【Abstract】 Objective To investigate the inhibitory effects of nanoparticle-mediated locked nucleic acid antigene(LNA antigene) on hepatitis B virus(HBV) in transgenic mouse model. Methods LNA antigene was conjugated with polylactic acid-glycolic acid(PLGA) to prepare LNA antigene nanoparticles(NPs), and particle size and surface charge were determined by nanoparticle tracking analysis(NTA) and Zeta potential measurement. Encapsulation efficiency, drug loading capacity, and uptake efficiency in HepG2 hepatocellular carcinoma cells were evaluated. Subsequently, the in vitro release characteristics of LNA anti gene NP and its inhibitory effects on the expressions of HBV DNA, HBsAg, and HBeAg in transgenic mice were analyzed. Results LNA antigene-NP was successfully prepared with a particle size ranging from 125 nm to 225 nm, and a lower surface charge compared to unmodified NPs. The encapsulation efficiency of LNA antigene-NP was 32%, and drug loading capacity was 8.448 mg. LNA antigene-NP exhibited significant cellular uptake efficiency in HepG2 hepatocellular carcinoma cells. The in vitro release studies demonstrated a sustained release over 24 days, with a final release rate approaching 90%. In the transgenic mouse model, LNA antigene-NP exhibited dose-dependent inhibition of HBV DNA, HBsAg, and HBeAg, with 5 kg/mg dose group achieving inhibition rates of about 40%, 60%, and 60% on day 7, respectively. Conclusion LNA antigene-NP can effectively inhibit HBV replication and antigen expression, which provides an experimental basis for further development of LNA-based anti-HBV therapies.更多还原