Basic Study
Copyright ©The Author(s) 2020. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Jun 7, 2020; 26(21): 2810-2820
Published online Jun 7, 2020. doi: 10.3748/wjg.v26.i21.2810
Gan Shen Fu Fang ameliorates liver fibrosis in vitro and in vivo by inhibiting the inflammatory response and extracellular signal-regulated kinase phosphorylation
Qing-Hong Du, Chu-Jun Zhang, Wei-Hong Li, Yan Mu, Ya Xu, Scott Lowe, Lin Han, Xue Yu, Shu-Yan Wang, Yu Li, Jian Li
Qing-Hong Du, Chu-Jun Zhang, Yan Mu, Ya Xu, Lin Han, Xue Yu, Shu-Yan Wang, Yu Li, Jian Li, Department of Histology and Embryology, School of Traditional Chinese Medicine, Beijing University of Chinese Medicine, Beijing 102488, China
Qing-Hong Du, Institute of Tibetan Medicine, University of Tibetan Medicine, Lhasa 850000, Tibet Autonomous Region, China
Wei-Hong Li, School of Nursing, Beijing University of Chinese Medicine, Beijing 102488, China
Scott Lowe, School of Molecular and Cellular Biology, University of Illinois, Urbana-Champaign, IL 61820, United States
Author contributions: Du QH and Li J designed the research; Du QH wrote this paper; Zhang CJ, Du QH, Li WH, Li J, Xu Y, Han L, Mu Y, Lowe S, Yu X and Wang SY performed the experiments; Li Y contributed new analytic tools; all authors approved the final version of the article.
Supported by the Innovation Team of the Beijing University of Chinese Medicine, No. 2019-JYB-TD-006; the National Natural Science Foundation of China, No. 81873099; Scientific Research Support Plan for the Construction of Doctoral Program of University of Tibetan Medicine.
Institutional review board statement: The study was reviewed and approved by the Institutional Review Board at Beijing University of Chinese Medicine.
Institutional animal care and use committee statement: All experimental procedures were conducted in accordance with the guidelines for the use of experimental animals and were approved by the Institutional Review Committee on Animal Care and Use at the Experimental Animal Centre of Beijing University of Chinese Medicine [certificate of conformity: SCXK (2012-0001)].
Conflict-of-interest statement: All other authors have nothing to disclose.
Data sharing statement: No additional data are available.
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 that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (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:
Corresponding author: Jian Li, PhD, Professor, Teacher, Department of Histology and Embryology, School of Traditional Chinese Medicine, Beijing University of Chinese Medicine, No.11 Bei San Huan Dong Road, Beijing 100029, China.
Received: February 17, 2020
Peer-review started: February 17, 2020
First decision: March 30, 2020
Revised: March 30, 2020
Accepted: April 27, 2020
Article in press: April 27, 2020
Published online: June 7, 2020
Research background

Liver fibrosis is a common healthy problem worldwide and there is still a lack of specific medicine. The Chinese herbal medicine,Gan Shen Fu Fang (GSFF) is composed of salvianolic acid B and diammonium glycyrrhizinate. In this study, we observe the effects of GSFF on liver fibrosis in vivo and in vitro in an attempt to provide some hope for the treatment.

Research motivation

During pre-clinical experiment, we found that GSFF could inhibit pseudo-lobule formation in common bile duct-ligated (CBDL) rats. However, the mechanisms remain unclear. Therefore, in this study, we aimed to observe the effects of GSFF on liver fibrosis in vivo and in vitro and determine whether GSFF-mediated alleviation of liver fibrosis is related to inflammation and the ERK signalling pathway.

Research objectives

The present study aimed to observe the effects of GSFF on liver fibrosis in vivo and in vitro and investigate the mechanism from the perspective of the inflammatory response and extracellular signal-regulated kinase (ERK) phosphorylation.

Research methods

CBDL rats were used for in vivo experiments. Hepatic stellate cells-T6(HSC-T6) cells were used for in vitro experiments. Hematoxylin and eosin and Masson staining, biochemical assays, hydroxyproline assays, enzyme-linked immunoasorbentassay and western blotting were performed to evaluate the degree of liver fibrosis, liver function, the inflammatory response and ERK phosphorylation. The CCK8 assay, immunofluorescence and western blotting were applied to test the effect of GSFF on HSC-T6 cell activation and determine whether GSFF had an effect on ERK phosphorylation in HSC-T6 cells.

Research results

GSFF improved liver function and inhibited liver fibrosis in CBDL rats after 3wk of treatment, as demonstrated by histological changes, hydroxyproline assays and collagen I concentrations. GSFF alleviated inflammatory cell infiltration and reduced the synthesis of pro-inflammatory cytokines (tumor necrosis factor-α and interlukin-1β) and NF-κB. In addition, GSFF decreased ERK phosphorylation. In vitro, GSFF inhibited the viability of HSC-T6 cells with and without transforming growth factor β1 stimulation and decreased the synthesis of collagen I. GSFF had the greatest effect at a concentration of 0.5 μmol/L. GSFF inhibited the expression of α-smooth muscle actin, a marker of HSC activation, in HSC-T6 cells. Consistent with the in vivo results, GSFF also inhibited the phosphorylation of ERK and downregulated the expression of NF-κB.

Research conclusions

GSFF inhibited liver fibrosis progression in vivo and HSC-T6 cell activation in vitro. These effects may be related to an alleviated inflammatory response and downregulated ERK phosphorylation.

Research perspectives

The definite anti-liver fibrosis effect and clear mechanism of GSFF provide hope for the treatment of liver fibrosis.