Abstract
Liver fibrosis is a consequence of chronic liver disease that can progress to liver cirrhosis or even hepatocarcinoma. Fuzheng Huayu (FZHY), a Chinese herbal formula, has been shown to exert anti-fibrotic effects. To better understand the molecular mechanisms underlying the anti-fibrotic effects of FZHY, we analyzed transcriptomic and proteomic combination profiles in CCl4-induced liver fibrosis in rats, which were treated with extracted FZHY powder (0.35 g·kg−1·d−1, ig) for 3 weeks. We showed that FZHY administration significantly improved liver function, alleviated hepatic inflammatory and fibrotic changes, and decreased the hydroxyproline content in the livers of CCl4-treated rats. When their liver tissues were examined using microarray and iTRAQ, we found 255 differentially expressed genes (fold change ≥1.5, P<0.05) and 499 differentially expressed proteins (fold change ≥1.2, P<0.05) in the FZHY and model groups. Functional annotation with DAVID (The Database for Annotation, Visualization and Integrated Discovery) showed that 15 enriched gene ontology terms, including drug metabolic process, response to extracellular stimulus, response to vitamins, arachidonic acid metabolic process, response to wounding, and oxidation reduction might be involved in the anti-fibrotic effects of FZHY; whereas KEGG pathway analysis revealed that eight enriched pathways, including arachidonic acid metabolism, retinol metabolism, metabolism of xenobiotics by cytochrome P450, and drug metabolism might also be involved. Moreover, the protein-protein interaction network demonstrated that 10 core genes/proteins overlapped, with Ugt2a3, Cyp2b1 and Cyp3a18 in retinol metabolism pathway overlapped to a higher degree. Compared to the model rats, the livers of FZHY-treated rats had significantly higher mRNA and protein expression levels of Ugt2a3, Cyp2b1 and Cyp3a18. Furthermore, the concentration of retinoic acid was significantly higher in the FZHY-treated rats compared with the model rats. The results suggest that the anti-fibrotic effects of FZHY emerge through multiple targets, multiple functions, and multiple pathways, including FZHY-regulated retinol metabolism, xenobiotic metabolism by cytochrome P450, and drug metabolism through up-regulated Ugt2a3, Cyp2b1, and Cyp3a18. These genes may play important anti-fibrotic roles in FZHY-treated rats.
Similar content being viewed by others
Log in or create a free account to read this content
Gain free access to this article, as well as selected content from this journal and more on nature.com
or
References
Su TH, Kao JH, Liu CJ . Molecular mechanism and treatment of viral hepatitis-related liver fibrosis. Int J Mol Sci 2014; 15: 10578–604.
Sebastiani G, Gkouvatsos K, Pantopoulos K . Chronic hepatitis C and liver fibrosis. World J Gastroenterol 2014; 20: 11033–53.
Czaja AJ . Hepatic inflammation and progressive liver fibrosis in chronic liver disease. World J Gastroenterol 2014; 20: 2515–32.
Elpek GÖ . Cellular and molecular mechanisms in the pathogenesis of liver fibrosis: An update. World J Gastroenterol 2014; 20: 7260–76.
Dong S, Su SB . Advances in mesenchymal stem cells combined with traditional Chinese medicine therapy for liver fibrosis. J Integr Med 2014; 12: 147–55.
Zhao CQ, Wu YQ, Xu LM . Curative effects of Fuzheng Huayu capsules on hepatic fibrosis and the functional mechanisms: a review. Zhong Xi Yi Jie He Xue Bao 2006; 4: 467–72.
Dong S, Chen QL, Feng Q, Hu YY, Liu P, Su SB . Study of molecular mechanism of Fuzheng-Huayu Formula improved rats liver fibrosis based on analysis of gene expression profile. China J Tradit Chin Med Pharm 2015; 30: 1812–7.
Dong S, Chen QL, Su SB . Curative effects of Fuzheng Huayu on liver fibrosis and cirrhosis: a meta-analysis. Evid Based Complement Alternat Med 2015; 2015: 125659.
Liu C, Jiang CM, Liu CH, Liu P, Hu YY . Effect of Fuzheng Huayu decoction on vascular endothelial growth factor secretion in hepatic stellate cells. Hepatobiliary Pancreat Dis Int 2002; 1: 207–10.
Wang QL, Tao YY, Shen L, Cui HY, Liu CH . Chinese herbal medicine Fuzheng Huayu recipe inhibits liver fibrosis by mediating the transforming growth factor-β1/Smads signaling pathway. Zhong Xi Yi Jie He Xue Bao 2012; 10: 561–8.
Wang Q, Du H, Li M, Li Y, Liu S, Gao P, et al. MAPK signal transduction pathway regulation: a novel mechanism of rat HSC-T6 cell apoptosis induced by FUZHENGHUAYU tablet. Evid Based Complement Alternat Med 2013; 2013: 368103.
Li H, Zhao L, Zhang B, Jiang Y, Wang X, Guo Y, et al. A network pharmacology approach to determine active compounds and action mechanisms of ge-gen-qin-lian decoction for treatment of type 2 diabetes. Evid Based Complement Alternat Med 2014; 2014: 495840.
Zhang GB, Li QY, Chen QL, Su SB . Network pharmacology: a new approach for Chinese herbal medicine research. Evid Based Complement Alternat Med 2013; 2013: 621423.
Cheng L, Pan GF, Zhang XD, Wang JL, Wang WD, Zhang JY, et al. Yindanxinnaotong, a Chinese compound medicine, synergistically attenuates atherosclerosis progress. Sci Rep 2015; 5: 12333.
Shi SH, Cai YP, Cai XJ, Zheng XY, Cao DS, Ye FQ, et al. A network pharmacology approach to understanding the mechanisms of action of traditional medicine: Bushenhuoxue formula for treatment of chronic kidney disease. PLoS One 2014; 9: e89123.
Li X, Wu L, Liu W, Jin Y, Chen Q, Wang L, et al. A network pharmacology study of Chinese medicine QiShenYiQi to reveal its underlying multi-compound, multi-target, multi-pathway mode of action. PLoS One 2014; 9: e95004.
An L, Feng F . Network pharmacology-based antioxidant effect study of zhi-zi-da-huang decoction for alcoholic liver disease. Evid Based Complement Alternat Med 2015; 2015: 492470.
Tang F, Tang Q, Tian Y, Fan Q, Huang Y, Tan X . Network pharmacology-based prediction of the active ingredients and potential targets of Mahuang Fuzi Xixin decoction for application to allergic rhinitis. J Ethnopharmacol 2015; 176: 402–12.
Zhang GB, Song YN, Chen QL, Dong S, Lu YY, Su MY, et al. Actions of Huangqi decoction against rat liver fibrosis: a gene expression profiling analysis. Chin Med 2015; 10: 39.
Song YN, Dong S, Wei B, Liu P, Zhang YY, Su SB . Metabolomic mechanisms of gypenoside against liver fibrosis in rats: An integrative analysis of proteomics and metabolomics data. PLoS One 2017; 12: e0173598.
Wang RQ, Mi HM, Li H, Zhao SX, Jia YH, Nan YM . Modulation of IKKβ/NF-κB and TGF-β1/Smad via Fuzheng Huayu recipe involves in prevention of nutritional steatohepatitis and fibrosis in mice. Iran J Basic Med Sci 2015; 18: 404–11.
Liu C, Liu P, Liu CH, Zhu XQ, Ji G . Effects of Fuzhenghuayu decoction on collagen synthesis of cultured hepatic stellate cells, hepatocytes and fibroblasts in rats. World J Gastroenterol 1998; 4: 548–9.
Lykkedegn S, Sorensen GL, Beck-Nielsen SS, Pilecki B, Duelund L, Marcussen N, et al. Vitamin D depletion in pregnancy decreases survival time, oxygen saturation, lung weight and body weight in preterm rat offspring. PLoS One 2016; 11: e0155203.
Pandit JJ, Handy JM . Science, Anaesthesia and animal studies: what is 'evidence'? Anaesthesia 2010; 65: 223–6.
Watanabe T, Suzuki T, Natsume M, Nakajima M, Narumi K, Hamada S, et al. Discrimination of genotoxic and non-genotoxic hepatocarcinogens by statistical analysis based on gene expression profiling in the mouse liver as determined by quantitative real-time PCR. Mutat Res 2012; 747: 164–75.
Crosas-Molist E, Fabregat I . Role of NADPH oxidases in the redox biology of liver fibrosis. Redox Biol 2015; 6: 106–11.
Lurie Y, Webb M, Cytter-Kuint R, Shteingart S, Lederkremer GZ . Non-invasive diagnosis of liver fibrosis and cirrhosis. World J Gastroenterol 2015; 21: 11567–83.
Crossan C, Tsochatzis EA, Longworth L, Gurusamy K, Davidson B, Rodríguez-Perálvarez M, et al. Cost-effectiveness of non-invasive methods for assessment and monitoring of liver fibrosis and cirrhosis in patients with chronic liver disease: systematic review and economic evaluation. Health Technol Assess 2015; 19: 1–409, v–vi.
Liu P, Hu YY, Liu C, Xu LM, Liu CH, Sun KW, et al. Multicenter clinical study on Fuzhenghuayu capsule against liver fibrosis due to chronic hepatitis B. World J Gastroenterol 2005; 11: 2892–9.
Abdel-Moneim AM, Al-Kahtani MA, El-Kersh MA, Al-Omair MA . Free radical-scavenging, anti-inflammatory/anti-fibrotic and hepatoprotective actions of taurine and silymarin against CCl4 induced rat liver damage. PLoS One 2015; 10: e0144509.
Ogaly HA, Eltablawy NA, El-Behairy AM, El-Hindi H, Abd-Elsalam RM . Hepatocyte growth factor mediates the antifibrogenic action of Ocimum bacilicum essential oil against CCl4-induced liver fibrosis in rats. Molecules 2015; 20: 13518–35.
Sun S, Dai J, Wang W, Cao H, Fang J, Hu YY, et al. Metabonomic evaluation of ZHENG differentiation and treatment by Fuzhenghuayu tablet in hepatitis-B-caused cirrhosis. Evid Based Complement Alternat Med 2012; 2012: 453503.
Ippolito DL, AbdulHameed MD, Tawa GJ, Baer CE, Permenter MG, McDyre BC, et al. Gene expression patterns associated with histopathology in toxic liver fibrosis. Toxicol Sci 2016; 149: 67–88.
Dong S, Chen QL, Song YN, Sun Y, Wei B, Li XY, et al. Mechanisms of CCl4-induced liver fibrosis with combined transcriptomic and proteomic analysis. J Toxicol Sci 2016; 41: 561–72.
Mohamed Suhaimi NA, Zhuo L . Imidazolium salt attenuates thioacetamide-induced liver fibrosis in mice by modulating inflammation and oxidative stress. Dig Liver Dis 2012; 44: 665–73.
He W, Shi F, Zhou ZW, Li B, Zhang K, Zhang X, et al. A bioinformatic and mechanistic study elicits the antifibrotic effect of ursolic acid through the attenuation of oxidative stress with the involvement of ERK, PI3K/Akt, and p38 MAPK signaling pathways in human hepatic stellate cells and rat liver. Drug Des Devel Ther 2015; 9: 3989–4104.
Domitrović R, Jakovac H, Marchesi VV, Blažeković B . Resolution of liver fibrosis by isoquinoline alkaloid berberine in CCl4-intoxicated mice is mediated by suppression of oxidative stress and upregulation of MMP-2 expression. J Med Food 2013; 16: 518–28.
Di Maso V, Mediavilla MG, Vascotto C, Lupo F, Baccarani U, Avellini C, et al. Transcriptional up-regulation of APE1/Ref-1 in hepatic tumor: role in hepatocytes resistance to oxidative stress and apoptosis. PLoS One 2015; 10: e0143289.
Teraoka R, Shimada T, Aburada M . The molecular mechanisms of the hepatoprotective effect of gomisin A against oxidative stress and inflammatory response in rats with carbon tetrachloride-induced acute liver injury. Biol Pharm Bull 2012; 35: 171–7.
Park S, Choi S, Lee MG, Lim C, Oh J . Retinol binding protein-albumin domain III fusion protein deactivates hepatic stellate cells. Mol Cell 2012; 34: 517–22.
Yi SH, Zhang Y, Tang D, Zhu L . Mechanical force and tensile strain activated hepatic stellate cells and inhibited retinol metabolism. Biotechnol Lett 2015; 37: 1141–52.
Miyazaki H, Takitani K, Koh M, Inoue A, Kishi K, Tamai H . Retinol status and expression of retinol-related proteins in methionine-choline deficient rats. J Nutr Sci Vitaminol (Tokyo) 2014; 60: 78–85.
De Paula TP, Ramalho A, Braulio VB . The effectiveness of relative dose response to retinol intake as an evaluation of vitamin A status of cirrhotic patients. J Hum Nutr Diet 2010; 23: 583–9.
Chang WT, Ker CG, Hung HC, Lee KT, Chen LS, Chiang HC, et al. Albumin and prealbumin may predict retinol status in patients with liver cirrhosis. Hepatogastroenterology 2008; 55: 1681–5.
Hisamori S, Tabata C, Kadokawa Y, Okoshi K, Tabata R, Mori A, et al. All-trans-retinoic acid ameliorates carbon tetrachloride-induced liver fibrosis in mice through modulating cytokine production. Liver Int 2008; 28: 1217–25.
Wang L, Potter JJ, Rennie-Tankersley L, Novitskiy G, Sipes J, Mezey E . Effects of retinoic acid on the development of liver fibrosis produced by carbon tetrachloride in mice. Biochem Biophys Acta 2007; 1772: 66–71.
Panebianco C, Oben JA, Vinciguerra M, Pazienza V . Senescence in hepatic stellate cells as a mechanism of liver fibrosis reversal: a putative synergy between retinoic acid and PPAR-gamma signalings. Clin Exp Med 2017; 17: 269–80.
Acknowledgements
This work was supported by Key Program of National Natural Science Foundation of China (81330084), National Science and Technology Major Project of China (2009ZX09311-003), Shanghai Municipal Science and Technology Commission Project (12401900401), and E-institutes of Shanghai Municipal Education Commission (E03008).
Author information
Authors and Affiliations
Corresponding authors
Additional information
Supplementary Table S1 is available on the website of Acta Pharmacologica Sinica.
Supplementary information
Supplementary Table S1
Pearson correlation analysis of gene expression and retinoic acid production (PDF 46 kb)
Rights and permissions
About this article
Cite this article
Dong, S., Cai, Ff., Chen, Ql. et al. Chinese herbal formula Fuzheng Huayu alleviates CCl4-induced liver fibrosis in rats: a transcriptomic and proteomic analysis. Acta Pharmacol Sin 39, 930–941 (2018). https://doi.org/10.1038/aps.2017.150
Received:
Accepted:
Published:
Issue date:
DOI: https://doi.org/10.1038/aps.2017.150
Keywords
This article is cited by
-
Differential proteomics for studying action mechanisms of traditional Chinese medicines
Chinese Medicine (2019)
-
New omic and network paradigms for deep understanding of therapeutic mechanisms for Fangji of traditional Chinese medicine
Acta Pharmacologica Sinica (2018)
