Basic Study
Copyright ©The Author(s) 2015. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Jul 21, 2015; 21(27): 8326-8339
Published online Jul 21, 2015. doi: 10.3748/wjg.v21.i27.8326
Givinostat inhibition of hepatic stellate cell proliferation and protein acetylation
Yu-Gang Wang, Ling Xu, Ting Wang, Jue Wei, Wen-Ying Meng, Na Wang, Min Shi
Yu-Gang Wang, Ling Xu, Ting Wang, Jue Wei, Wen-Ying Meng, Na Wang, Min Shi, Department of Gastroenterology, Shanghai Tongren Hospital, Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200336, China
Author contributions: Wang YG, Shi M, and Xu L designed the study; Wang YG, Wang N, Shi M, Wei J, Xu L, and Wang T carried out the study; Wang N, Shi M, Wei J, and Wang T contributed new reagents/analytic tools; Wei J, Wang YG, and Meng WY analyzed the data; and Shi M and Wang YG wrote the paper.
Supported by Shanghai Municipal Health Bureau Key Disciplines Grant, No. ZK2012A05; Bootstrap Class Project Foundation of the Science and Technology Commission of Shanghai Municipality, No. 14411973700; and Shanghai Municipal Health Bureau, No. 20134100.
Institutional review board statement: In vivo experiments were approved by the Shanghai Tongren Hospital affiliated to Shanghai Jiao Tong University School of Medicine Ethics Committee.
Institutional animal care and use committee statement: In vivo experiments were approved by the Shanghai Tongren Hospital affiliated to Shanghai Jiao Tong University School of Medicine Institutional animal care and use committee.
Conflict-of-interest statement: The authors declare no conflicts of interest.
Data sharing statement: No additional unpublished data are available.
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: Min Shi, MD, PhD, Professor, Department of Gastroenterology, Shanghai Tongren Hospital, Affiliated to Shanghai Jiao Tong University School of Medicine, 1111 Xianxia Street, Shanghai 200336, China. shimingdyx@yeah.net
Telephone: +86-21-62909911 Fax: +86-21-62906478
Received: January 19, 2015
Peer-review started: January 20, 2015
First decision: February 26, 2015
Revised: March 2, 2015
Accepted: May 21, 2015
Article in press: May 21, 2015
Published online: July 21, 2015
Processing time: 183 Days and 14.7 Hours
Abstract

AIM: To explore the effect of the histone deacetylase inhibitor givinostat on proteins related to regulation of hepatic stellate cell proliferation.

METHODS: The cell counting kit-8 assay and flow cytometry were used to observe changes in proliferation, apoptosis, and cell cycle in hepatic stellate cells treated with givinostat. Western blot was used to observe expression changes in p21, p57, CDK4, CDK6, cyclinD1, caspase-3, and caspase-9 in hepatic stellate cells exposed to givinostat. The scratch assay was used to analyze the effect of givinostat on cell migration. Effects of givinostat on the reactive oxygen species profile, mitochondrial membrane potential, and mitochondrial permeability transition pore opening in JS-1 cells were observed by laser confocal microscopy.

RESULTS: Givinostat significantly inhibited JS-1 cell proliferation and promoted cell apoptosis, leading to cell cycle arrest in G0/G1 phases. Treatment with givinostat downregulated protein expression of CDK4, CDK6, and cyclin D1, whereas expression of p21 and p57 was significantly increased. The givinostat-induced apoptosis of hepatic stellate cells was mainly mediated through p38 and extracellular signal-regulated kinase 1/2. Givinostat treatment increased intracellular reactive oxygen species production, decreased mitochondrial membrane potential, and promoted mitochondrial permeability transition pore opening. Acetylation of superoxide dismutase (acetyl K68) and nuclear factor-κB p65 (acetyl K310) was upregulated, while there was no change in protein expression. Moreover, the notable beneficial effect of givinostat on liver fibrosis was also confirmed in the mouse models.

CONCLUSION: Givinostat has antifibrotic activities via regulating the acetylation of nuclear factor-κB and superoxide dismutase 2, thus inhibiting hepatic stellate cell proliferation and inducing apoptosis.

Keywords: Givinostat; Hepatic stellate cells; Histone deacetylase inhibitor; Nuclear factor-κB; Superoxide dismutase

Core tip: There is currently no effective therapeutic treatment for liver fibrosis. Inhibition of hepatic stellate cell activation and proliferation or induction of apoptosis is the mainstream strategy for the treatment of liver fibrosis. This study demonstrates that a histone deacetylase inhibitor, givinostat, has antifibrotic activities both in vivo and in vitro, which might be achieved by regulating the acetylation of nuclear factor-κB and superoxide dismutase, thus stimulating oxidative stress, activating mitochondrial pathways, inhibiting hepatic stellate cell proliferation, and inducing apoptosis. These results may provide new directions and evidence in the research and development of novel drugs for liver fibrosis.