Targeted delivery of siRNA against hepatitis B virus by preS1 peptide molecular ligand
Targeted delivery of siRNA against hepatitis B virus by preS1 peptide molecular ligand
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preS1肽分子配体靶向递送针对乙型肝炎病毒的siRNA
DOI：
10.1111/hepr.12189
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发表时间：
2014-08-01
影响因子：
4.2
通讯作者：
Chen, Weixian
中科院分区：
文献类型：
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作者：
Huang, Wenjuan;Li, Xia;Chen, Weixian
目的：对于慢性B型肝炎病毒（HBV）感染，目前的治疗效果有限。RNA干扰病毒特异性基因已成为一种潜在的抗病毒机制。然而，仍有待开发合适的递送载体。本研究通过构建一种靶向肝细胞的siRNA载体，在体内外研究其对HBV相关肝病的治疗作用，为HBV相关肝病的治疗提供新的思路。为了验证体内抗病毒功效，通过皮下注射建立HBV病毒血症动物模型。将HepG 2 2.2.15肿瘤细胞接种于裸鼠中。通过凝胶电泳检测对siRNA迁移的最小阻滞作用，以确认组合和最佳比例。半定量酶联免疫吸附试验检测细胞表面B型肝炎表面抗原（HBsAg）水平，实时定量聚合酶链反应检测RNA水平，免疫印迹和免疫组化检测蛋白水平。结论：PreS 1 - 9Arg-siRNA分子偶联物在体内外均能有效抑制HBsAg和HBV DNA的产生，且无肝毒性。
Aim: For chronic hepatitis B virus (HBV) infection, the effects of current therapies are limited. RNA interference of virus-specific genes has emerged as a potential antiviral mechanism. However, a suitable delivery vector is still to be developed. We studied a novel vector transferring siRNA targeting hepatic cells in vivo and in vitro in order to find a new way to cure HBV-related live diseases.Methods: The preS1-9Arg ligand was used to deliver siRNA to HepG2 and to HepG2 2.2.15 cells. To validate the antiviral efficacy in vivo, a HBV viremic animal model was established by s.c. inoculation of HepG2 2.2.15 tumor cells in nude mice. The minimal retardation effect on the migration of siRNA was detected by gel electrophoresis to confirm the combination and the optimal ratio. Hepatitis B surface antigen (HBsAg) levels were detected by semiquantitatively enzyme-linked immunosorbent assay RNA levels were quantified with quantitative real-time polymerase chain reaction and protein levels were determined with immunoblots and immunohistochemistry.Results: PreS1-9Arg peptide strongly combined and transferred siRNA into HepG2 cells. PreS1-9Arg-siRNA molecular conjugate effectively reduced the production of HBsAg and HBV DNA without liver toxicity in vitro and in vivo.Conclusion: The results indicated that preS1-9Arg may be a potential novel vector to deliver siRNA targeting liver cells.