Basic Study
Copyright ©The Author(s) 2015. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Apr 14, 2015; 21(14): 4184-4194
Published online Apr 14, 2015. doi: 10.3748/wjg.v21.i14.4184
Early activated hepatic stellate cell-derived molecules reverse acute hepatic injury
Wen-Ju Chang, Lu-Jun Song, Tuo Yi, Kun-Tang Shen, Hong-Shan Wang, Xiao-Dong Gao, Min Li, Jian-Min Xu, Wei-Xin Niu, Xin-Yu Qin
Wen-Ju Chang, Lu-Jun Song, Tuo Yi, Kun-Tang Shen, Hong-Shan Wang, Xiao-Dong Gao, Min Li, Jian-Min Xu, Wei-Xin Niu, Xin-Yu Qin, Department of General Surgery, Zhongshan Hospital, Institute of General Surgery, Fudan University, Shanghai 200032, China
Author contributions: Chang WJ and Qin XY designed the research; Chang WJ, Song LJ, and Yi T performed the research; Shen KT, Wang HS, Gao XD, and Li M contributed new reagents or analytic tools; Xu JM and Niu WX analyzed the data; Chang WJ and Yi T wrote the paper; Chang WJ, Song LJ, and Yi T contributed equally to this work.
Supported by Doctoral Fund of the Ministry of Education, No. 2013007110041; Young Investigator Funding of Zhongshan Hospital, No. 2014ZSQN37.
Ethics approval: The study was reviewed and approved by the Zhongshan Hospital Review Board.
Institutional animal care and use committee: All procedures involving animals were reviewed and approved by the Institutional Animal Care and Use Committee of the Zhongshan Hospital, Fudan University (IACUC protocol number: Y2013-0019).
Conflict-of-interest: The authors have no conflict of interest related to the manuscript.
Data sharing: No additional 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: Xin-Yu Qin, MD, Department of General Surgery, Zhongshan Hospital, Institute of General Surgery, Fudan University, No. 220 Handan Road, Shanghai 200032, China. qin.xinyu@zs-hospital.sh.cn
Telephone: +86-21-64041990 Fax: +86-21-64037224
Received: September 26, 2014
Peer-review started: September 29, 2014
First decision: October 29, 2014
Revised: December 16, 2014
Accepted: January 16, 2015
Article in press: January 16, 2015
Published online: April 14, 2015
Processing time: 200 Days and 12.6 Hours
Abstract

AIM: To test whether hepatic stellate cells (HSCs) at different activation stages play different roles in acetaminophen (APAP)-induced acute liver injury (ALI).

METHODS: HSCs were isolated from mouse liver and cultured in vitro. Morphological changes of initiation HSCs [HSCs (5d)] and perpetuation HSCs [HSCs (p3)] were observed by immunofluorescence and transmission electron microscopy. The protective effects of HSC-derived molecules, cell lysates and HSC-conditioned medium (HSC-CM) were tested in vivo by survival and histopathological analyses. Liver injury was determined by measuring aminotransferase levels in the serum and by histologic examination of tissue sections under a light microscope. Additionally, to determine the molecular mediators of the observed protective effects of initiation HSCs, we examined HSC-CM using a high-density protein array.

RESULTS: HSCs (5d) and HSCs (p3) had different morphological and phenotypic traits. HSCs (5d) presented a star-shaped appearance with expressing α-SMA at non-uniform levels between cells. However, HSCs (p3) evolved into myofibroblast-like cells without lipid droplets and expressed a uniform and higher level of α-SMA. HSC-CM (5d), but not HSC-CM (p3), provided a significant survival benefit and showed a dramatic reduction of hepatocellular necrosis and panlobular leukocyte infiltrates in mice exposed to APAP. However, this protective effect was abrogated at higher cell masses, indicating a therapeutic window of effectiveness. Furthermore, the protein array screen revealed that HSC-CM (5d) was composed of many chemokines and growth factors that correlated with inflammatory inhibition and therapeutic activity. When compared with HSC-CM (p3), higher levels of monocyte chemoattractant protein-1, macrophage inflammatory protein-1γ, hepatocyte growth factor, interleukin-10, and matrix metalloproteinase-2, but lower levels of stem cell factor and Fas-Ligand were observed in HSC-CM (5d).

CONCLUSION: These data indicated that initiation HSCs and perpetuation HSCs were different in morphology and protein expression, and provided the first experimental evidence of the potential medical value of initiation HSC-derived molecules in the treatment of ALI.

Keywords: Hepatic stellate cells; Acute liver injury; Initiation and perpetuation

Core tip: In this study, we isolated hepatic stellate cells (HSCs) from mice by in situ perfusion of the liver and created primary and secondary cultures in plastic tissue culture dishes. Then, we observed different morphologies and phenotypes between initiation HSCs and perpetuation HSCs and described the first use of molecules secreted from HSCs in acetaminophen-induced acute liver injury. Initiation HSC-derived molecules showed hepatocyte-protective effects. Our findings provide novel insight into the mechanisms of HSCs in liver injury therapy. Whether the potential value of initiation HSC-derived molecular therapy is derived from the effect of a single cytokine or a combination of cytokines should be explored in future.