Original Article
Copyright ©2012 Baishideng Publishing Group Co., Limited. All rights reserved.
World J Gastroenterol. Jul 28, 2012; 18(28): 3696-3704
Published online Jul 28, 2012. doi: 10.3748/wjg.v18.i28.3696
Lentiviral vector-mediated down-regulation of IL-17A receptor in hepatic stellate cells results in decreased secretion of IL-6
Sheng-Chu Zhang, Yi-Hu Zheng, Pan-Pan Yu, Tan Hooi Min, Fu-Xiang Yu, Chao Ye, Yuan-Kang Xie, Qi-Yu Zhang
Sheng-Chu Zhang, Yi-Hu Zheng, Pan-Pan Yu, Fu-Xiang Yu, Yuan-Kang Xie, Qi-Yu Zhang, Department of General Surgery, The First Affiliated Hospital of Wenzhou Medical College, Wenzhou 325000, Zhejiang Province, China
Tan Hooi Min, Stem Cell Center, Lund University, 22184 Lund, Sweden
Chao Ye, Department of Infectious Disease, Binjiang Hospital of Hangzhou, Hangzhou 310051, Zhejiang Province, China
Author contributions: Zhang SC, Yu PP, Min TH and Yu FX performed the majority of experiments; Ye C and Xie YK provided analytical tools and were also involved in editing the manuscript; Zhang QY and Zheng YH designed the study and wrote the manuscript.
Supported by The Zhejiang Extremely Key Subject of Surgery; and The Wenzhou Key Laboratory Project in Surgery
Correspondence to: Qi-Yu Zhang, Professor, Department of General Surgery, The First Affiliated Hospital of Wenzhou Medical College, Wenzhou 325000, Zhejiang Province, China. zgzhqy@126.com
Telephone: +86-577-88069639 Fax: +86-577-88069703
Received: October 5, 2011
Revised: April 4, 2012
Accepted: May 6, 2012
Published online: July 28, 2012
Abstract

AIM: To investigate the mechanism of interleukin (IL)-6 secretion through blocking the IL-17A/IL-17A receptor (IL-17RA) signaling pathway with a short hairpin RNA (shRNA) in hepatic stellate cells (HSCs) in vitro.

METHODS: HSCs were derived from the livers of adult male Sprague-Dawley rats. IL-6 expression was evaluated using real-time quantitative polymerase chain reaction and enzyme linked immunosorbent assay. The phosphorylation activity of p38 mitogen activated protein kinases (MAPK) and extracellular regulated protein kinases (ERK) 1/2 upon induction by IL-17A and suppression by IL-17RA shRNA were examined using Western blotting.

RESULTS: IL-6 expression induced by IL-17A was significantly increased compared to control in HSCs (P < 0.01 in a dose-dependent manner). Suppression of IL-17RA using lentiviral-mediated shRNA inhibited IL-6 expression induced by IL-17A compared to group with only IL-17A treatment (1.44 ± 0.17 vs 4.07 ± 0.43, P < 0.01). IL-17A induced rapid phosphorylation of p38 MAPK and ERK1/2 after 5 min exposure, and showed the strongest levels of phosphorylation of p38 MAPK and ERK1/2 at 15 min in IL-17A-treated HSCs. IL-6 mRNA expression induced by IL-17A (100 ng/mL) for 3 h exposure was inhibited by preincubation with specific inhibitors of p38 MAPK (SB-203580) and ERK1/2 (PD-98059) compared to groups without inhibitors preincubation (1.67 ± 0.24, 2.01 ± 0.10 vs 4.08 ± 0.59, P < 0.01). Moreover, Lentiviral-mediated IL-17RA shRNA 1 inhibited IL-17A-induced IL-6 mRNA expression compared to random shRNA in HSCs (1.44 ± 0.17 vs 3.98 ± 0.68, P < 0.01). Lentiviral-mediated IL-17RA shRNA 1 inhibited phosphorylation of p38 MAPK and ERK1/2 induced by 15 min IL-17A (100 ng/mL) exposure.

CONCLUSION: Down-regulation of the IL-17RA receptor by shRNA decreased IL-6 expression induced by IL-17A via p38 MAPK and ERK1/2 phosphorylation in HSCs. Suppression of IL-17RA expression may be a strategy to reduce the inflammatory response induced by IL-17A in the liver.

Keywords: Interleukin 17A, Interleukin 6, Hepatic stellate cells, Liver fibrosis