Vitamin E-coupled nanoparticles-delivered siRNA inhibits expression of hepatitis C virus core protein <italic style="font-style: italic">in vitro</italic>

Xiaoping LI; Yan YAN; Hong CHEN; Weixian CHEN; Ying HUANG

doi:10.16016/j.1000-5404.201705123

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Vitamin E-coupled nanoparticles-delivered siRNA inhibits expression of hepatitis C virus core protein in vitro

Basic Medicine | 更新时间：2026-01-09

- Vitamin E-coupled nanoparticles-delivered siRNA inhibits expression of hepatitis C virus core protein in vitro
- Journal of Third Military Medical University Vol. 39, Issue 20, Pages: 1980-1985(2017)
- 作者机构：
  
  1.400010 重庆, 重庆医科大学附属第二医院: 科研处
  2.400010 重庆, 重庆医科大学附属第二医院: 检验科
  3.400010 重庆, 重庆医科大学附属第二医院: 感染科
- 作者简介：
- 基金信息：
  
  the General Program of National Natural Science Foundation of China(81171628)
- DOI：10.16016/j.1000-5404.201705123
  CLC： R373.21;R394.6;R977.25
- Received：31 May 2017，
  
  Revised：2017-07-18，
  
  Published：30 October 2017
- 稿件说明：
移动端阅览
Xiaoping LI, Yan YAN, Hong CHEN, et al. Vitamin E-coupled nanoparticles-delivered siRNA inhibits expression of hepatitis C virus core protein in vitro[J]. Journal of Third Military Medical University, 2017, 39(20): 1980-1985.
DOI：

Xiaoping LI, Yan YAN, Hong CHEN, et al. Vitamin E-coupled nanoparticles-delivered siRNA inhibits expression of hepatitis C virus core protein in vitro[J]. Journal of Third Military Medical University, 2017, 39(20): 1980-1985. DOI： 10.16016/j.1000-5404.201705123.

摘要

目的

研究携带针对丙型肝炎病毒(hepatitis C virus

HCV)RNA的小干扰RNA(siRNA)的维生素E(Vitamin E)脂质体纳米颗粒在体外靶向抑制HCV复制的作用。

方法

采取“薄膜水化法”制备Vitamin E脂质体纳米颗粒，应用激光粒度分析仪测量纳米颗粒VE-DC/siRNA的颗粒大小及zeta电位，应用透射电子显微镜观察VE-DC形态及分散性。以Huh7.5.1细胞株作为载体。CCK-8法检测纳米颗粒的细胞毒性，琼脂糖凝胶电泳检测纳米颗粒携带siRNA的血清稳定性，蛋白印迹技术和免疫荧光技术检测该siRNA纳米颗粒的转染效率及对HCV的抑制作用。

结果

Vitamin E脂质体纳米颗粒形态呈球形，粒度分布为(125.5±9.0) nm，zeta电位为(39.1±4.8) mV，分散均一。相较同浓度商品化脂质体，VE-DC/siRNA对细胞的毒性相对较小。血清稳定性实验显示，VE-DC/siRNA在24 h后siRNA无降解，较裸siRNA稳定性显著提高。携带siRNA的Vitamin E脂质体纳米颗粒可以较高效率进入Huh7.5.1细胞内，并抑制HCV核心蛋白的表达。

结论

Vitamin E脂质体纳米颗粒可以转运siRNA至Huh7.5.1细胞，并抑制HCV核心蛋白的表达。

Abstract

Objective

To study the inhibitory effect of small interfering RNAs (siRNAs) targeting hepatitis C virus (HCV) delivered by vitamin E-coupled nanoparticles on the viral replication

in vitro

Methods

Vitamin E-coupled nanoparticles were prepared by thin-film hydration method. The size and surface charge (zeta potential) of the VE-DC/siRNAs nanoparticles were determined with a laser particle size analyzer. The homogeneity and dispersivity of the nanoparticles were analyzed by transmission electron microscopy. CCK-8 assay was used to assess the cytotoxicity of VE-DC/siRNAs. The stability of the nanoparticles was determined by agarose gel electrophoresis. The transfection efficiency and the inhibition of HCV was analyzed with Western blotting and immunofluorescent staining.

Results

The VE-DC/siRNAs nanoparticles were in spherical shape and homogeneous distribution

with their size distribution of 125.5±9.0 nm and zeta potential of 39.1±4.8 mV. These nanoparticles were characterized for their lower cytotoxicity and higher stability. The CCK-8 assay data showed that cell viability was higher for VE-DC/siRNAs treated cells than those treated with commercial liposomes at the same concentration. In serum

siRNA was intact in the VE-DC/siRNAs nanoparticles after 24 h

which was degraded in the naked siRNA within 6 h. TheVE-DC/siRNAs nanoparticles were efficiently taken up by hepatoma cells(huh7.5.1-HCV

huh7.5.1-core) observed under fluorescence microscope. Western blot analysis detected decreased levels of HCV nonstructural protein 3 (NS3) and HCV-core protein in the VE-DC/siRNAs treated hepatoma cells.

Conclusion

Targeted delivery of siRNAs by vitamin E-coupled nanoparticles effectively inhibits HCV replication and core protein production

suggesting that it may potentially use as delivery vehicle for target therapy.

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Keywords

references

KOHLI A, SHAFFER A, SHERMAN A, et al. Treatment of hepatitis C: a systematic review[J]. JAMA, 2014, 312(6): 631-640. DOI: 10.1001/jama.2014.7085.

GONZ LEZ-REIMERS E, QUINTERO-PLATT G, RODR GUEZ-GASPAR M, et al. Liver steatosis in hepatitis C patients[J]. World J Hepatol, 2015, 7(10): 1337-1346. DOI: 10.4254/wjh.v7.i10.1337.

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Vitamin E-coupled nanoparticles-delivered siRNA inhibits expression of hepatitis C virus core protein in vitro

DOI：10.16016/j.1000-5404.201705123

摘要

Abstract

关键词

Keywords

references