Basic Study
Copyright ©The Author(s) 2017. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Aug 7, 2017; 23(29): 5333-5344
Published online Aug 7, 2017. doi: 10.3748/wjg.v23.i29.5333
Salvianolic acid B protects hepatocytes from H2O2 injury by stabilizing the lysosomal membrane
Xiao-Feng Yan, Pei Zhao, Dong-Yan Ma, Yi-Lu Jiang, Jiao-Jiao Luo, Liu Liu, Xiao-Ling Wang
Xiao-Feng Yan, Pei Zhao, Xiao-Ling Wang, Department of Biology, School of Basic Medical Science, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China
Dong-Yan Ma, Women and Infant Hospital of Zhengzhou, Zhengzhou 450000, Henan Province, China
Yi-Lu Jiang, Jiao-Jiao Luo, Liu Liu, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China
Author contributions: Wang XL and Yan XF designed the research; Yan XF, Zhao P, Ma DY, Jiang YL, Luo JJ and Liu L performed the research; Yan XF, Zhao P and Ma DY analyzed the data; Wang XL wrote the paper.
Supported by National Natural Science Funds of China, No. 81503367; and the Budget Research Project of Shanghai Education Commission, No. 2014YSN03 and No. 2014YSN22.
Institutional review board statement: This study was reviewed and approved by the Shanghai University of Traditional Chinese Medicine Institutional Review Board (No. 201409011).
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 Shanghai University of Traditional Chinese Medicine [License number: SCXK (Shanghai) 2014-0008].
Conflict-of-interest statement: The authors declare that there are no conflicts of interest.
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: Dr. Xiao-Ling Wang, Department of Biology, School of Basic Medical Science, Shanghai University of Traditional Chinese Medicine, 1200 Cailun Road, Pudong, Shanghai 201203, China. wangxiaoling1033@shutcm.edu.cn
Telephone: +86-21-51322585
Received: January 17, 2017
Peer-review started: January 19, 2017
First decision: February 23, 2017
Revised: March 27, 2017
Accepted: May 9, 2017
Article in press: May 9, 2017
Published online: August 7, 2017
Abstract
AIM

To investigate the capability of salvianolic acid B (Sal B) to protect hepatocytes from hydrogen peroxide (H2O2)/carbon tetrachloride (CCl4)-induced lysosomal membrane permeabilization.

METHODS

Cell Counting Kit-8 assay was used to measure cell viability. Apoptosis and death were assayed through flow cytometry. BrdU incorporation was used to detect cell proliferation. Serum alanine aminotransferase activity and liver malondialdehyde (MDA) content were measured. Liver histopathological changes were evaluated using hematoxylin-eosin staining. Lysosomal membrane permeability was detected with LysoTracker Green-labeled probes and acridine orange staining. The levels of protein carbonyl content (PCC), cathepsins (Cat)B/D, and lysosome-associated membrane protein 1 (LAMP1) were evaluated through western blotting. Cytosol CatB activity analysis was performed with chemiluminescence detection. The mRNA level of LAMP1 was evaluated through quantitative real-time polymerase chain reaction.

RESULTS

Results indicated that H2O2 induced cell injury/death. Sal B attenuated H2O2-induced cell apoptosis and death, restored the inhibition of proliferation, decreased the amount of PCC, and stabilized the lysosome membrane by increasing the LAMP1 protein level and antagonizing CatB/D leakage into the cytosol. CCl4 also triggered hepatocyte death. Furthermore, Sal B effectively rescued hepatocytes by increasing LAMP1 expression and by reducing lysosomal enzyme translocation to the cytosol.

CONCLUSION

Sal B protected mouse embryonic hepatocytes from H2O2/CCl4-induced injury/death by stabilizing the lysosomal membrane.

Keywords: Lysosomal membrane permeabilization, Injury, Salvianolic acid B, Hepatocyte

Core tip: Lysosomal membrane permeabilization leads to the release of luminal contents, such as proteases and protons, into the cytosol, resulting in cell death. Salvianolic acid B (Sal B), a water-soluble compound extracted from Salvia miltiorrhiza, exerts a cell protective effect by anti-oxidation. In the present study, we elucidate that Sal B protects hepatocytes from H2O2/CCl4 -induced injury/death by stabilizing the lysosomal membrane.