Basic Study
Copyright ©The Author(s) 2017. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Aug 28, 2017; 23(32): 5904-5912
Published online Aug 28, 2017. doi: 10.3748/wjg.v23.i32.5904
Effects and mechanism of adenovirus-mediated phosphatase and tension homologue deleted on chromosome ten gene on collagen deposition in rat liver fibrosis
Shu-Rui Xie, Jun-Yan An, Li-Bo Zheng, Xiao-Xia Huo, Jian Guo, David Shih, Xiao-Lan Zhang
Shu-Rui Xie, Jun-Yan An, Li-Bo Zheng, Xiao-Xia Huo, Jian Guo, Xiao-Lan Zhang, Department of Gastroenterology, The Second Hospital of Hebei Medical University, Hebei Key Laboratory of Gastroenterology, Hebei Institute of Gastroenterology, Shijiazhuang 050000, Hebei Province, China
Shu-Rui Xie, Department of Gastroenterology, Xingtai People's Hospital, Xingtai 054031, Hebei Province, China
David Shih, Inflammatory Bowel and Immunobiology Research Institute, F. Widjaja Foundation, Cedars-Sinai Medical Center, Los Angeles, CA 90048, United States
Author contributions: Xie SR and An JY contributed equally to this work; Xie SR and An JY performed the majority of the experiments; Zheng LB and Guo J assisted with various experiments and helped to analyze the data; Huo XX and Zhang XL drafted and edited the manuscript; Shih D performed critical revision of the manuscript.
Supported by the National Natural Science Foundation of China, No. 30872513.
Institutional review board statement: This study was reviewed and approved by the Second Hospital of Hebei Medical University Institutional Review Board.
Institutional animal care and use committee statement: All procedures involving animals were reviewed and approved by the Institutional Animal Care and Use Committee of the Experimental Animal Center of Hebei Medical University (No. 911102).
Conflict-of-interest statement: We declare that there are no conflicts of interest to disclose.
Data sharing statement: Technical appendix, statistical code, and dataset available from the corresponding author at xiaolanzh@126.com. Participants gave informed consent for data sharing.
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: Xiao-Lan Zhang, MD, PhD, Professor, Department of Gastroenterology, The Second Hospital of Hebei Medical University, Hebei Key Laboratory of Gastroenterology, Hebei Institute of Gastroenterology, 215 West Heping Road, Shijiazhuang 050000, Hebei Province, China. xiaolanzh@hb2h.com
Telephone: +86-311-66007370 Fax: +86-311-66007370
Received: January 26, 2017
Peer-review started: February 4, 2017
First decision: April 21, 2017
Revised: June 19, 2017
Accepted: July 22, 2017
Article in press: July 24, 2017
Published online: August 28, 2017
Processing time: 213 Days and 8.5 Hours
Abstract
AIM

To evaluate the effects of phosphatase and tension homologue deleted on chromosome ten (PTEN) gene on collagen metabolism in hepatic fibrosis and the underlying mechanisms.

METHODS

Rat primary hepatic stellate cells (HSCs) and human LX-2 cells were transfected with adenovirus containing cDNA constructs encoding wild-type PTEN (Ad-PTEN), PTEN mutant G129E gene (Ad-G129E), and RNA interference constructs targeting the PTEN sequence PTEN short hairpin RNA to up-regulate and down-regulate the expression of PTEN. HSCs were assayed using fluorescent microscopy, real-time polymerase chain reaction, and western blotting. Moreover, a CCl4-induced rat hepatic fibrosis model was established to investigate the in vivo effects. Hematoxylin and eosin, and Masson’s trichrome were used to assess the histological changes. The expression of collagen I and III was assessed using immunohistochemistry and western blot analysis.

RESULTS

Elevated expression of PTEN gene reduced serum levels of alanine transaminase and aspartate transaminase, decreased collagen deposition in the liver, and reduced hepatocyte necrosis. In contrast, knockdown of PTEN expression had an opposite effect, such as increased collagen deposition in the liver, and was molecularly characterized by the increased expression of matrix metalloproteinase (MMP)-13 (P < 0.01) and MMP-2 (P < 0.01), as well as decreased expression of the tissue inhibitor of metalloproteinase (TIMP)-1 (P < 0.01) and TIMP-2 (P < 0.01).

CONCLUSION

These data indicated that gene therapy using recombinant adenovirus encoding PTEN might be a novel way of treating hepatic fibrosis.

Keywords: Collagen metabolism; Hepatic stellate cells; Phosphatase and tension homologue deleted on chromosome ten; PTEN; Gene therapy; Hepatic fibrosis

Core tip: Phosphatase and tension homologue deleted on chromosome ten (PTEN) has a negative relation with the activation and proliferation of hepatic stellate cells (HSCs), which is the central event in liver fibrogenesis as HSCs are the major source of collagens and matrix metalloproteinases in fibrotic liver. In this study, adenoviruses containing cDNA constructs encoding wild-type PTEN (Ad-PTEN) and PTEN mutant G129E gene (Ad-G129E) were constructed to over-express the PTEN gene in both rat primary HSCs and human LX-2 cells as well as in the CCl4-induced rat liver fibrosis model. The adenovirus-mediated over-expression of the PTEN gene attenuated extracellular matrix (ECM) synthesis (collagens I and III) and promoted ECM degradation, representing a possible novel anti-fibrosis therapy.