Basic Study
Copyright ©The Author(s) 2015. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Aug 14, 2015; 21(30): 9079-9092
Published online Aug 14, 2015. doi: 10.3748/wjg.v21.i30.9079
Protective effect of Salvia miltiorrhiza and Carthamus tinctorius extract against lipopolysaccharide-induced liver injury
Li-Na Gao, Kuo Yan, Yuan-Lu Cui, Guan-Wei Fan, Yue-Fei Wang
Li-Na Gao, Kuo Yan, Yuan-Lu Cui, Guan-Wei Fan, Yue-Fei Wang, Research Center of Traditional Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China
Li-Na Gao, Kuo Yan, Yuan-Lu Cui, Guan-Wei Fan, Tianjin State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China
Yue-Fei Wang, Research and Development Center of TCM, Tianjin International Joint Academy of Biotechnology and Medicine, Tianjin 300457, China
Author contributions: Gao LN and Yan K performed the majority of experiments; Gao LN and Cui YL performed the literature research; Gao LN, Fan GW, Wang YF and Yan K performed the data acquisition and data analysis; Gao LN drafted the manuscript and made critical revisions related to important intellectual content of the manuscript; Cui YL designed the whole study and reviewed the manuscript; all authors read and approved the final manuscript.
Supported by National Natural Science Foundation of China, No. 81173469 and No. 81273891; and the Key New Drug Creation and Manufacturing Program, No. 2012ZX09304007.
Institutional review board statement: The study was reviewed and approved by the Ethics Committee of Tianjin University of Traditional Chinese Medicine Institutional Review Board.
Institutional animal care and use committee statement: All procedures involving animals were reviewed and approved by the Institutional Animal Care and Use Committee of the Ethics Committee of Tianjin University of Traditional Chinese Medicine (IACUC protocol number: TCM-2009-037-E10).
Conflict-of-interest statement: Gao LN and Yan K are students of Tianjin University of Traditional Chinese Medicine. Cui YL, Fan GW and Wang YF are employees of Tianjin University of Traditional Chinese Medicine. All authors declare no conflict of interest.
Data sharing statement: Technical appendix, statistical code, and dataset available from the corresponding author at cuiyl@tju.edu.cn. Participants gave informed consent for 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: Yuan-Lu Cui, Professor, Research Center of Traditional Chinese Medicine, Tianjin University of Traditional Chinese Medicine, No. 88 YuQuan Road, Nankai District, Tianjin 300193, China. cuiyl@tju.edu.cn
Telephone: +86-22-59596170 Fax: +86-22-59596170
Received: March 8, 2015
Peer-review started: March 11, 2015
First decision: April 24, 2015
Revised: May 9, 2015
Accepted: June 9, 2015
Article in press: June 10, 2015
Published online: August 14, 2015
Abstract

AIM: To investigate the hepatoprotective effects and mechanisms of an extract of Salvia miltiorrhiza and Carthamus tinctorius in vivo.

METHODS: C57BL/6J mice were randomly assigned to five groups and intraperitoneally administered 0.9% saline, Salvia miltiorrhiza and Carthamus tinctorius extract [Danhong injection (DHI), 0.75 and 3 g/kg mixed extract] or reduced glutathione for injection (RGI, 300 mg/kg) for 30 min before exposure to lipopolysaccharide (LPS, 16 mg/kg). After intraperitoneal LPS stimulation for 90 min or 6 h, the mice were sacrificed by ether anaesthesia, and serum and liver samples were collected. Histological analysis (H&E) and terminal deoxynucleotidyl transferase-mediated dUTP nick end-labelling (TUNEL) staining were performed. Alanine transferase (ALT), aspartate transaminase (AST), total bilirubin (TBil), glutathione-S-transferase (GST), malondialdehyde (MDA), tumour necrosis factor (TNF)-α, interleukin (IL)-6, and caspase-3 levels were measured. Bax, Bcl-2, P-IκBα, IκBα, P-NF-κB p65, and NF-κB p65 protein levels were determined by Western blot. TNF-α, IL-6, caspase-3, Bax and Bcl-2 mRNA expression was measured by real-time reverse transcription-polymerase chain reaction (RT-PCR).

RESULTS: Hematoxylin-eosin staining and TUNEL results suggested that DHI (3 g/kg) treatment alleviated inflammatory and apoptotic (P < 0.01) injury in the liver of mice. DHI treatment dose-dependently blunted the abnormal changes in biochemical parameters such as ALT (72.53 ± 2.83 for 3 g/kg, P < 0.01), AST (76.97 ± 5.00 for 3 g/kg, P < 0.01), TBil (1.17 ± 0.10 for 3 g/kg, P < 0.01), MDA (0.81 ± 0.36 for 3 g/kg, P < 0.01), and GST (358.86 ± 12.09 for 3 g/kg, P < 0.01). Moreover, DHI (3 g/kg) remarkably decreased LPS-induced protein expression of TNF-α (340.55 ± 10.18 for 3 g/kg, P < 0.01), IL-6 (261.34 ± 10.18 for 3 g/kg, P < 0.01), and enzyme activity of caspase-3 (0.93 ± 0.029 for 3 g/kg, P < 0.01). The LPS-induced mRNA expression of TNF-α, IL-6 and caspase-3 was also decreased by DHI. Western blot analysis revealed that DHI antagonised LPS-stimulated decrease of Bcl-2 and increase of Bax protein expression. Furthermore, DHI inhibited LPS-induced IκBα and NF-κB p65 phosphorylation.

CONCLUSION: DHI may be a multi-function protectant against acute hepatic injury in mice through its anti-inflammatory, anti-oxidative and anti-apoptotic activities.

Keywords: Salvia miltiorrhiza, Carthamus tinctorius, Apoptosis, Anti-inflammatory, Antioxidant, Acute liver injury

Core tip:Salvia miltiorrhiza and Carthamus tinctorius extract [Danhong injection (DHI)] effectively protected against hepatic injury. DHI intervention significantly reduced alanine transferase, aspartate transaminase, total bilirubin, malondialdehyde, glutathione-S-transferase, tumour necrosis factor-α, interleukin-6, and caspase-3 levels in an lipopolysaccharide (LPS)-induced mouse model of acute liver injury. Moreover, DHI antagonised LPS-induced Bcl-2 and Bax expression and inhibited IκBα and nuclear factor-κB p65 phosphorylation. These findings suggest that Salvia miltiorrhiza and Carthamus tinctorius extract (such as DHI) acts as a multi-function protectant against acute hepatic injury in mice through its anti-inflammatory, anti-oxidative and anti-apoptotic activities.