Basic Study
Copyright ©The Author(s) 2018. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Clin Cases. Aug 16, 2018; 6(8): 183-191
Published online Aug 16, 2018. doi: 10.12998/wjcc.v6.i8.183
Antiviral effects of hepatitis B virus S gene-specific anti-gene locked nucleic acid in transgenic mice
Shu-Rong Xiao, Gui-Dan Xu, Wu-Jun Wei, Bin Peng, Yi-Bin Deng
Shu-Rong Xiao, Gui-Dan Xu, Wu-Jun Wei, Bin Peng, Yi-Bin Deng, Department of Medical Laboratory Science, the Affiliated Hospital of Youjiang Medical College for Nationalities, Baise 533000, Guangxi Zhuang Autonomous Region, China
Yi-Bin Deng, Department of Hepatobiliary Disease Center, Guangxi Clinic Medicine Research, Baise 533000, Guangxi Zhuang Autonomous Region, China
Author contributions: Xiao SR and Deng YB conceived the study, analyzed the data, drafted the manuscript; Xu GD and Wei WJ helped revise the manuscript critically for important intellectual content; Peng B collect the data; Deng YB helped design the study.
Supported by National Natural Science Foundation of China, No. 81460123; Guangxi Graduate Innovation Program, No. 201601005; and Guangxi Clinic Medicine Research Center of Hepatobiliary Disease, No. AD17129025.
Institutional review board statement: This study was approved by the Affiliated Hospital of Youjiang Medical College for Nationalities, Baise, China.
Institutional animal care and use committee statement: All procedures involving animals were reviewed and approved by the Animal Ethics and Welfare Committee of Affiliated Hospital of Youjiang Medical College for Nationalities.
Conflict-of-interest statement: The authors declare that they have no competing interests.
ARRIVE guidelines statement: The authors have read the ARRIVE guidelines, and the manuscript was prepared and revised according to the ARRIVE guidelines.
Open-Access: This article is an open-access article which was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/
Correspondence to: Yi-Bin Deng, PhD, Professor, Department of Medical Laboratory Science, the Affiliated Hospital of Youjiang Medical College for Nationalities, 18 Zhongshan 2nd Road, Baise 533000, Guangxi Zhuang Autonomous Region, China. enbin0776@sina.com
Telephone: +86-776-2852592 Fax: +86-776-2852592
Received: January 24, 2018
Peer-review started: February 2, 2018
First decision: March 7, 2018
Revised: March 29, 2018
Accepted: June 7, 2018
Article in press: June 8, 2018
Published online: August 16, 2018
Abstract
AIM

To assess the antiviral effects of hepatitis B virus (HBV) S gene-specific anti-gene locked nucleic acid (LNA) in transgenic mice.

METHODS

Thirty HBV transgenic mice were acclimatized to laboratory conditions and positive for serum HBV surface antigen (HBsAg) and HBV DNA, were randomly divided into 5 groups (n = 7), including negative control (blank control, unrelated sequence control), positive control (lamivudine, anti-sense-LNA), and anti-gene-LNA experimental group. LNA was injected into transgenic mice by tail vein while lamivudine was administered by gavage. Serum HBV DNA and HBsAg levels were determined by fluorescence-based PCR and enzyme-linked immune sorbent assay, respectively. HBV S gene expression amounts were assessed by reverse transcription polymerase chain reaction. Positive rates of HBsAg in liver cells were evaluated immunohistochemistry.

RESULTS

Average rate reductions of HBsAg after treatment on the 3rd, 5th, and 7th days were 32.34%, 45.96%, and 59.15%, respectively. The inhibitory effect of anti-gene-LNA on serum HBsAg peaked on day 7, with statistically significant differences compared with pre-treatment (0.96 ± 0.18 vs 2.35 ± 0.33, P < 0.05) and control values (P < 0.05 for all). Average reduction rates of HBV DNA on the 3rd, 5th, and 7th days were 38.55%, 50.95%, and 62.26%, respectively. This inhibitory effect peaked on the 7th day after treatment with anti-gene-LNA, with statistically significant differences compared with pre-treatment (4.17 ± 1.29 vs 11.05 ± 1.25, P < 0.05) and control values (P < 0.05 for all). The mRNA levels of the HBV S gene (P < 0.05 for all) and rates of HBsAg positive liver cells (P < 0.05 for all) were significantly reduced compared with the control groups. Liver and kidney function, and histology showed no abnormalities.

CONCLUSION

Anti-gene-LNA targeting the S gene of HBV displays strong inhibitory effects on HBV in transgenic mice, providing theoretical and experimental bases for gene therapy in HBV.

Keywords: Anti-gene therapy, Hepatitis B virus, Locked nucleic acid, Hepatitis B, Transgenic mice, Anti-sense-therapy

Core tip: We assess the antiviral effects of hepatitis B virus (HBV) S gene-specific anti-gene locked nucleic acid (LNA) in transgenic mice, to provide an experimental basis for gene therapy in patients with Chronic B-related Hepatitis. The inhibitory effect of anti-gene-LNA on serum HBV surface antigen (HBsAg) and HBV DNA peaked on day 7, with statistically significant differences compared with pre-treatment and control values. The mRNA levels of the HBVS gene and rates of HBsAg positive liver cells were significantly reduced compared with the control groups.