Basic Study
Copyright ©The Author(s) 2016. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Feb 14, 2016; 22(6): 2092-2103
Published online Feb 14, 2016. doi: 10.3748/wjg.v22.i6.2092
Human urokinase-type plasminogen activator gene-modified bone marrow-derived mesenchymal stem cells attenuate liver fibrosis in rats by down-regulating the Wnt signaling pathway
Zhi-Gang Ma, Xiao-Dan Lv, Ling-Ling Zhan, Lan Chen, Qi-Yuan Zou, Ji-Qiao Xiang, Jiao-Li Qin, Wei-Wei Zhang, Zhao-Jing Zeng, Hui Jin, Hai-Xing Jiang, Xiao-Ping Lv
Zhi-Gang Ma, Lan Chen, Qi-Yuan Zou, Ji-Qiao Xiang, Jiao-Li Qin, Wei-Wei Zhang, Zhao-Jing Zeng, Hui Jin, Hai-Xing Jiang, Xiao-Ping Lv, Department of Gastroenterology, the First Affiliated Hospital of Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China
Xiao-Dan Lv, Ling-Ling Zhan, Department of Clinical Experimental Medicine, the First Affiliated Hospital of Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China
Author contributions: Lv XP and Zhan LL designed and supervised the research; Ma ZG, Lv XD, Chen L and Zou QY performed the experiments; Ma ZG, Xiang JQ, Qin JL, Zhang WW, Zeng ZJ, Jin H and Jiang HX acquired and analyzed the data; Ma ZG drafted and wrote the manuscript; Lv XP, Zhan LL and Chen L revised the manuscript for important intellectual content; all the authors have read and approved the final version to be published.
Supported by National Natural Science Foundation of China, No. 81460114; Natural Science Foundation of Guangxi Zhuang Autonomous Region, No. 1355005-3-2 and No. 2012GXNSFAA053143; Chinese Traditional Medicine Science Foundation of Guangxi Zhuang Autonomous Region, No. GZPT1238; and Science Foundation of Guangxi Department of Education, No. 201203YB036 and No. 2013LX031.
Institutional review board statement: The study was reviewed and approved by the First Affiliated Hospital of Guangxi Medical University Ethical Review Committee.
Institutional animal care and use committee statement: All procedures involving animals were reviewed and approved by the Institutional Animal Care and Use Committee of Guangxi Medical University.
Conflict-of-interest statement: The authors declare no conflict of interests.
Data sharing statement: 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: Xiao-Ping Lv, Professor, Department of Gastroenterology, the First Affiliated Hospital of Guangxi Medical University, No. 6 Shuangyong Road, Nanning 530021, Guangxi Zhuang Autonomous Region, China. lxxp58@hotmail.com
Telephone: +86-771-3277211 Fax: +86-771-3277285
Received: August 2, 2015
Peer-review started: August 3, 2015
First decision: September 9, 2015
Revised: September 27, 2015
Accepted: November 24, 2015
Article in press: November 24, 2015
Published online: February 14, 2016
Processing time: 173 Days and 19.3 Hours
Abstract

AIM: To evaluate the therapeutic effects of bone marrow-derived mesenchymal stem cells (BMSCs) with human urokinase-type plasminogen activator (uPA) on liver fibrosis, and to investigate the mechanism of gene therapy.

METHODS: BMSCs transfected with adenovirus-mediated human urokinase plasminogen activator (Ad-uPA) were transplanted into rats with CCl4-induced liver fibrosis. All rats were sacrificed after 8 wk, and their serum and liver tissue were collected for biochemical, histopathologic, and molecular analyzes. The degree of liver fibrosis was assessed by hematoxylin and eosin or Masson’s staining. Western blot and quantitative reverse transcription-polymerase chain reaction were used to determine protein and mRNA expression levels.

RESULTS: Serum levels of alanine aminotransferase, aminotransferase, total bilirubin, hyaluronic acid, laminin, and procollagen type III were markedly decreased, whereas the levels of serum albumin were increased by uPA gene modified BMSCs treatment. Histopathology revealed that chronic CCl4-treatment resulted in significant fibrosis while uPA gene modified BMSCs treatment significantly reversed fibrosis. By quantitatively analysing the fibrosis area of liver tissue using Masson staining in different groups of animals, we found that model animals with CCl4-induced liver fibrosis had the largest fibrotic area (16.69% ± 1.30%), while fibrotic area was significantly decreased by BMSCs treatment (12.38% ± 2.27%) and was further reduced by uPA-BMSCs treatment (8.31% ± 1.21%). Both protein and mRNA expression of β-catenin, Wnt4 and Wnt5a was down-regulated in liver tissues following uPA gene modified BMSCs treatment when compared with the model animals.

CONCLUSION: Transplantation of uPA gene modified BMSCs suppressed liver fibrosis and ameliorated liver function and may be a new approach to treating liver fibrosis. Furthermore, treatment with uPA gene modified BMSCs also resulted in a decrease in expression of molecules of the Wnt signaling pathway.

Keywords: Bone marrow-derived mesenchymal stem cells; Liver fibrosis; Urokinase plasminogen activator; Wnt signaling pathway

Core tip: It has been confirmed that urokinase plasminogen activator (uPA) has a protective effect in liver fibrosis. Bone marrow-derived mesenchymal stem cells (BMSCs) have been discovered to provide effective therapy for liver fibrosis. Therefore, the present study was designed to investigate the therapeutic effects of uPA gene modified BMSCs in a rat model of CCl4-induced liver fibrosis, and the impact on the Wnt signaling pathway which is involved in the pathogenesis of liver fibrosis. uPA gene modified BMSCs can suppress liver fibrosis and ameliorate liver function. Furthermore, it also resulted in down-regulation of molecules of the Wnt signaling pathway and may be a new approach to treating liver fibrosis.