Basic Study
Copyright ©The Author(s) 2015. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. May 14, 2015; 21(18): 5473-5481
Published online May 14, 2015. doi: 10.3748/wjg.v21.i18.5473
Dihydromyricetin alleviates carbon tetrachloride-induced acute liver injury via JNK-dependent mechanism in mice
Jun Xie, Jie Liu, Tu-Ming Chen, Qing Lan, Qing-Yu Zhang, Bin Liu, Dong Dai, Wei-Dong Zhang, Li-Ping Hu, Run-Zhi Zhu
Jun Xie, Wei-Dong Zhang, Li-Ping Hu, Run-Zhi Zhu, Clinical Research Center, Xuyi People’s Hospital, Xuyi 211700, Jiangsu Province, China
Jie Liu, Tu-Ming Chen, Qing-Yu Zhang, Bin Liu, Dong Dai, Run-Zhi Zhu, Department of Hepatobiliary Surgery of Affiliated Hospital of Guangdong Medical College, Zhanjiang Key Laboratory of Hepatobiliary Diseases, Zhanjiang 524001, Guangdong Province, China
Qing Lan, Department of Stomatology, Affiliated Hospital of Guangdong Medical College, Zhanjiang 524001, Guangdong Province, China
Author contributions: Xie J, Liu J and Chen TM contributed equally to this work; Xie J, Liu J and Chen TM performed the majority of experiments; Lan Q, Liu B and Zhang QY participated in the mouse sample collection; Dai D and Zhang WD participated in the preparation of the paraffin sections; Zhu RZ and Hu LP designed these experiments and guided their implementation.
Supported by Initial Fund of Guangdong Medical College, No. XB1338; the Medical Research Fund of Guangdong Province, No. B2014306; and the Research Fund of Guangdong Medical College, No. M2013024.
Ethics approval: The protocol of this study was approved by the Committee on the Ethics of Animal Experiments of Guangdong Medical College (Permit Number: SYXK 2008-0007).
Institutional animal care and use committee: All procedures involving animals were reviewed and approved by the Institutional Animal Care and Use Committee of the Guangdong Medical College (IACUC protocol number: AP3324).
Conflict-of-interest: Zhu RZ has received research funding from Guangdong Medical College; Liu J has received research funding from the public health and family planning council of Guangdong Province and Guangdong Medical College; Xie J, Zhang WD and Hu LP are employees of Xuyi People’s Hospital; Liu J, Chen TM, Lan Q, Dai D and Zhu RZ are employees of Affiliated Hospital of Guangdong Medical College; all authors declare that there is no conflict of interest in this work.
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:
Correspondence to: Run-Zhi Zhu, PhD, Clinical Research Center, Xuyi People’s Hospital, No. 3 East Huaihe Road, Xucheng Town, Xuyi 211700, Jiangsu Province, China.
Telephone: +86-759-2387596 Fax: +86-759-2387596
Received: November 7, 2014
Peer-review started: November 8, 2014
First decision: December 11, 2014
Revised: December 31, 2014
Accepted: February 12, 2015
Article in press: February 13, 2015
Published online: May 14, 2015
Processing time: 192 Days and 0.8 Hours

AIM: To assess the effects of dihydromyricetin (DHM) as a hepatoprotective candidate in reducing hepatic injury and accelerating hepatocyte proliferation after carbon tetrachloride (CCl4) treatment.

METHODS: C57 BL/6 mice were used in this study. Mice were orally administered with DHM (150 mg/kg) for 4 d after CCl4 treatment. Serum and liver tissue samples were collected on days 1, 2, 3, 5 and 7 after CCl4 treatment. The anti-inflammatory effect of DHM was assessed directly by hepatic histology detection and indirectly by serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), albumin, and superoxide dismutase (SOD). Inflammatory cytokines, such as interleukin (IL)-1β, IL-6 and tumor necrosis factor-α (TNF-α), were detected using ELISA kits. Proliferating cell nuclear antigen (PCNA) staining was used to evaluate the role of DHM in promoting hepatocyte proliferation. Hepatocyte apoptosis was measured by TUNEL assay. Furthermore, apoptosis proteins Caspases-3, 6, 8, and 9 were detected by Western blot. SP600125 were used to confirm whether DHM regulated liver regeneration through JNK/TNF-α pathways.

RESULTS: DHM showed a strong anti-inflammatory effect on CCl4-induced liver injury in mice. DHM could significantly decrease serum ALT, AST, IL-1β, IL-6 and TNF-α and increase serum albumin, SOD and liver SOD compared to the control group after CCl4 treatment (P < 0.05). PCNA results indicated that DHM could significantly increase the number of PCNA positive cells compared to the control (348.9 ± 56.0 vs 107.1 ± 31.4, P < 0.01). TUNEL assay showed that DHM dramatically reduced the number of apoptotic cells after CCl4 treatment compared to the control (365.4 ± 99.4 vs 90.5±13.8, P < 0.01). Caspase activity detection showed that DHM could reduce the activities of Caspases- 8, 3, 6 and 9 compared to the control (P < 0.05). The results of Western blot showed that DHM increased the expression of JNK and decreased TNF-α expression. However, DHM could not affect TNF-α expression after SP600125 treatment. Furthermore, DHM could significantly improve the survival rate of acute liver failure (ALF) mice (73.3% vs 20.0%, P < 0.0001), and SP600125 could inhibit the effect of DHM.

CONCLUSION: These findings demonstrate that DHM alleviates CCl4-induced liver injury, suggesting that DHM is a promising candidate for reversing liver injury and ALF.

Keywords: Dihydromyricetin, Liver regeneration, Tumor necrosis factor-α

Core tip: Our research confirmed that dihydromyricetin (DHM) plays an anti-inflammatory role in the carbon tetrachloride (CCl4) induced acute liver injury mice. It was shown that DHM could alleviate CCl4-induced acute liver injury by reducing apoptosis and accelerating proliferation of hepatocytes. Furthermore, DHM treatment up-regulated Jun kinase expression in liver tissue, and the mice which were injected with SP600125 could not survive in the acute liver failure induced by lethal dose CCl4 injection.