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Chinese Medicine
Published in: Chinese Medicine 1/2022

Open Access 01-12-2022 | Research

Revealing potential anti-fibrotic mechanism of Ganxianfang formula based on RNA sequence

Authors: Zongyi Liu, Huanyu Xiang, Dejuan Xiang, Shuang Xiao, Hongyan Xiang, Jing Xiao, Hong Ren, Peng Hu, Huabao Liu, Mingli Peng

Published in: Chinese Medicine | Issue 1/2022

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Abstract

Background

Ganxianfang (GXF) formula as a traditional Chinese medicine (TCM) is used for liver fibrosis in clinical practice while its mechanism is unclear. The aim of this study is to explore the molecular mechanism of GXF against CCl4-induced liver fibrosis rats.

Methods

Detected the main compounds of GXF by UPLC-MS/MS. Evaluated the efficacy of GXF (1.58, 3.15, 4.73 g/kg/day) and Fuzheng Huayu (FZHY, positive control, 0.47 g/kg/day) through serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) levels and histopathological changes. Explored the underlying mechanisms by integrating our total liver RNA sequencing (RNA-seq) data with recent liver single-cell sequencing (scRNA-seq) studies. Verified potential pharmacodynamic substances of GXF by hepatic stellate cell (HSC)-T6 line.

Results

Main compounds were identified in GXF by UPLC-MS/MS, including baicalin, wogonoside and matrine etc. With GXF-high dose treatment, the elevation of ALT and AST induced by CCl4 were significantly reduced, and the protective effect of GXF-high dose treatment was better than FZHY. Liver histopathological changes were alleviated by GXF-high dose treatment, the ISHAK scoring showed the incidence of liver cirrhosis (F5/F6) decreased from 76.5 to 55.6%. The results of liver hydroxyproline content were consistent with the histopathological changes. RNA-seq analysis revealed the differential genes (DEGs) were mainly enriched in ECM-receptor interaction and chemokine signaling pathway. GXF effectively inhibited collagen deposition and significantly downregulated CCL2 to inhibit the recruitment of macrophages in liver tissue. Integrating scRNA-seq data revealed that GXF effectively inhibited the expansion of scar-associated Trem2+CD9+ macrophages subpopulation and PDGFRα+PDGFRβ+ scar-producing myofibroblasts in the damaged liver, and remodeled the fibrotic niche via regulation of ligand-receptor interactions including TGFβ/EGFR, PDGFB/PDGFRα, and TNFSF12/TNFRSF12a signaling. In vitro experiments demonstrated that baicalin, matrine and hesperidin in GXF inhibited the activation of hepatic stellate cells.

Conclusions

This study clarified the potential anti-fibrotic effects and molecular mechanism of GXF in CCl4-induced liver fibrosis rats, which deserves further promotion and application.
Appendix
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Literature
1.
Kisseleva T, Brenner D. Molecular and cellular mechanisms of liver fibrosis and its regression. Nat Rev Gastroenterol Hepatol. 2021;18:151–66.PubMedCrossRef
2.
3.
4.
5.
Dong S, Su S. Advances in mesenchymal stem cells combined with traditional Chinese medicine therapy for liver fibrosis. J Integr Med. 2014;12:147–55.PubMedCrossRef
6.
Zhang L, Schuppan D. Traditional Chinese Medicine (TCM) for fibrotic liver disease: hope and hype. J Hepatol. 2014;61:166–8.PubMedCrossRef
7.
Luk JM, Wang X, Liu P, Wong K-F, Chan K-L, Tong Y, et al. Traditional Chinese herbal medicines for treatment of liver fibrosis and cancer: from laboratory discovery to clinical evaluation. Liver Int. 2007;27:879–90.PubMedCrossRef
8.
Xu L, Liu P. Guidelines for diagnosis and treatment of hepatic fibrosis with integrated traditional Chinese and Western medicine (2019 edition). J Integr Med. 2020;18:203–13.PubMedCrossRef
9.
Liu H, Lv J, Zhao Z, Xiong A, Tan Y, Glenn JS, et al. Fuzhenghuayu decoction ameliorates hepatic fibrosis by attenuating experimental sinusoidal capillarization and liver angiogenesis. Sci Rep. 2019;9:18719.PubMedPubMedCentralCrossRef
10.
Zhang M, Liu H, Huang K, Peng Y, Tao Y, Zhao C, et al. Fuzheng Huayu recipe prevented and treated CCl4-Induced mice liver fibrosis through regulating polarization and chemotaxis of intrahepatic macrophages via CCL2 and CX3CL1. Evid Based Complementary Altern Med. 2020;2020:8591892.CrossRef
11.
Fu Y, Xu B, Huang S, Luo X, Deng X, Luo S, et al. Baicalin prevents LPS-induced activation of TLR4/NF-κB p65 pathway and inflammation in mice via inhibiting the expression of CD14. Acta Pharmacol Sin. 2021;42:88–96.PubMedCrossRef
12.
Yao L, Wang S, Wei P, Bao K, Yuan W, Wang X, et al. Huangqi-Fangfeng protects against allergic airway remodeling through inhibiting epithelial–mesenchymal transition process in mice via regulating epithelial derived TGF-β1. Phytomedicine. 2019;64:153076.PubMedCrossRef
13.
Chen L, Lv D, Wang D, Chen X, Zhu Z, Cao Y, et al. A novel strategy of profiling the mechanism of herbal medicines by combining network pharmacology with plasma concentration determination and affinity constant measurement. Mol BioSyst. 2016;12:3347–56.PubMedCrossRef
14.
Kang J-W, Hong J-M, Lee S-M. Melatonin enhances mitophagy and mitochondrial biogenesis in rats with carbon tetrachloride-induced liver fibrosis. J Pineal Res. 2016;60:383–93.PubMedCrossRef
15.
Ogaly HA, Eltablawy NA, Abd-Elsalam RM. Antifibrogenic influence of Mentha piperita L. essential oil against CCl 4-induced liver fibrosis in rats. Oxid Med Cell Longev. 2018;2018:1–15.CrossRef
16.
Liu J, Kong D, Qiu J, Xie Y, Lu Z, Zhou C, et al. Praziquantel ameliorates CCl 4-induced liver fibrosis in mice by inhibiting TGF-β/Smad signalling via up-regulating Smad7 in hepatic stellate cells. Br J Pharmacol. 2019;176:4666–80.PubMedPubMedCentralCrossRef
17.
Xiong X, Kuang H, Ansari S, Liu T, Gong J, Wang S, et al. Landscape of intercellular crosstalk in healthy and NASH liver revealed by single-cell secretome gene analysis. Mol Cell. 2019;75:644-660.e5.PubMedPubMedCentralCrossRef
18.
Ramachandran P, Matchett KP, Dobie R, Wilson-Kanamori JR, Henderson NC. Single-cell technologies in hepatology: new insights into liver biology and disease pathogenesis. Nat Rev Gastroenterol Hepatol. 2020;17:457–72.PubMedCrossRef
19.
Ramachandran P, Dobie R, Wilson-Kanamori JR, Dora EF, Henderson BEP, Luu NT, et al. Resolving the fibrotic niche of human liver cirrhosis at single-cell level. Nature. 2019;575:512–8.PubMedPubMedCentralCrossRef
20.
Ishak’ K, Baptista A, Grootes JD, Gudat F, Denk H, Desmet V, et al. Histological grading and staging of chronic hepatitis. J Hepatol. 1995;22:696–9.CrossRef
21.
Mortazavi A, Williams BA, McCue K, Schaeffer L, Wold B. Mapping and quantifying mammalian transcriptomes by RNA-Seq. Nat Methods. 2008;5:621–8.PubMedCrossRef
22.
Liao Y, Smyth GK, Shi W. featureCounts: an efficient general purpose program for assigning sequence reads to genomic features. Bioinformatics. 2014;30:923–30.PubMedCrossRef
23.
Bray NL, Pimentel H, Melsted P, Pachter L. Near-optimal probabilistic RNA-seq quantification. Nat Biotechnol. 2016;34:525–7.PubMedCrossRef
24.
25.
Li H. Advances in anti hepatic fibrotic therapy with Traditional Chinese Medicine herbal formula. J Ethnopharmacol. 2020;251:112442.PubMedCrossRef
26.
Zhao T, Mao L, Yu Z, Hui Y, Feng H, Wang X, et al. Therapeutic potential of bicyclol in liver diseases: lessons from a synthetic drug based on herbal derivative in traditional Chinese medicine. Int Immunopharmacol. 2021;91:107308.PubMedCrossRef
27.
Dong S, Zhong Y, Lu W, Li G, Jiang H, Mao B. Baicalin inhibits lipopolysaccharide-induced inflammation through signaling NF-κB pathway in HBE16 airway epithelial cells. Inflammation. 2015;38:1493–501.PubMedCrossRef
28.
Chen Y, Yang Y, Wang F, Yang X, Yao F, Ming K, et al. Antiviral effect of baicalin phospholipid complex against duck hepatitis A virus type 1. Poult Sci. 2018;97:2722–32.PubMedCrossRef
29.
Suo Z, Liu Y, Ferreri M, Zhang T, Liu Z, Mu X, et al. Impact of matrine on inflammation related factors in rat intestinal microvascular endothelial cells. J Ethnopharmacol. 2009;125:404–9.PubMedCrossRef
30.
Sun D, Wang J, Yang N, Ma H. Matrine suppresses airway inflammation by downregulating SOCS3 expression via inhibition of NF-κB signaling in airway epithelial cells and asthmatic mice. Biochem Biophys Res Commun. 2016;477:83–90.PubMedCrossRef
31.
Ali AM, Gabbar MA, Abdel-Twab SM, Fahmy EM, Ebaid H, Alhazza IM, et al. Antidiabetic potency, antioxidant effects, and mode of actions of Citrus reticulata fruit peel hydroethanolic extract, hesperidin, and quercetin in nicotinamide/streptozotocin-induced Wistar diabetic rats. Oxid Med Cell Longev. 2020;2020:1730492.PubMedPubMedCentralCrossRef
32.
Mao Q-Q, Zhong X-M, Feng C-R, Pan A-J, Li Z-Y, Huang Z. Protective effects of paeoniflorin against glutamate-induced neurotoxicity in PC12 cells via antioxidant mechanisms and Ca2+ antagonism. Cell Mol Neurobiol. 2010;30:1059–66.PubMedCrossRef
33.
Zhou Y, Wu R, Cai F-F, Zhou W-J, Lu Y-Y, Zhang H, et al. Xiaoyaosan decoction alleviated rat liver fibrosis via the TGFβ/Smad and Akt/FoxO3 signaling pathways based on network pharmacology analysis. J Ethnopharmacol. 2021;264:113021.PubMedCrossRef
34.
Matyas C, Haskó G, Liaudet L, Trojnar E, Pacher P. Interplay of cardiovascular mediators, oxidative stress and inflammation in liver disease and its complications. Nat Rev Cardiol. 2021;18:117–35.PubMedCrossRef
35.
36.
Wen Y, Lambrecht J, Ju C, Tacke F. Hepatic macrophages in liver homeostasis and diseases-diversity, plasticity and therapeutic opportunities. Cell Mol Immunol. 2021;18:45–56.PubMedCrossRef
37.
Sahin H, Trautwein C, Wasmuth HE. Functional role of chemokines in liver disease models. Nat Rev Gastroenterol Hepatol. 2010;7:682–90.PubMedCrossRef
38.
Mandrekar P, Ambade A, Lim A, Szabo G, Catalano D. An essential role for monocyte chemoattractant protein-1 in alcoholic liver injury: regulation of proinflammatory cytokines and hepatic steatosis in mice. Hepatology. 2011;54:2185–97.PubMedCrossRef
39.
Baeck C, Wehr A, Karlmark KR, Heymann F, Vucur M, Gassler N, et al. Pharmacological inhibition of the chemokine CCL2 (MCP-1) diminishes liver macrophage infiltration and steatohepatitis in chronic hepatic injury. Gut. 2012;61:416–26.PubMedCrossRef
40.
Pellicoro A, Ramachandran P, Iredale JP, Fallowfield JA. Liver fibrosis and repair: immune regulation of wound healing in a solid organ. Nat Rev Immunol. 2014;14:181–94.PubMedCrossRef
41.
Krenkel O, Tacke F. Liver macrophages in tissue homeostasis and disease. Nat Rev Immunol. 2017;17:306–21.PubMedCrossRef
42.
Wehr A, Baeck C, Heymann F, Niemietz PM, Hammerich L, Martin C, et al. Chemokine receptor CXCR6-dependent hepatic NK T cell accumulation promotes inflammation and liver fibrosis. J Immunol. 2013;190:5226–36.PubMedCrossRef
43.
Castro-Sánchez S, García-Yagüe ÁJ, Kügler S, Lastres-Becker I. CX3CR1-deficient microglia shows impaired signalling of the transcription factor NRF2: implications in tauopathies. Redox Biol. 2019;22:101118.PubMedPubMedCentralCrossRef
44.
Karlmark KR, Zimmermann HW, Roderburg C, Gassler N, Wasmuth HE, Luedde T, et al. The fractalkine receptor CX3CR1 protects against liver fibrosis by controlling differentiation and survival of infiltrating hepatic monocytes. Hepatology. 2010;52:1769–82.PubMedCrossRef
45.
Wu R, Nakatsu G, Zhang X, Yu J. Pathophysiological mechanisms and therapeutic potentials of macrophages in non-alcoholic steatohepatitis. Expert Opin Ther Targets. 2016;20:615–26.PubMedCrossRef
46.
Krenkel O, Puengel T, Govaere O, Abdallah AT, Mossanen JC, Kohlhepp M, et al. Therapeutic inhibition of inflammatory monocyte recruitment reduces steatohepatitis and liver fibrosis: metabolic liver disease. Hepatology. 2018;67:1270–83.PubMedCrossRef
47.
Minutti CM, Modak RV, Macdonald F, Li F, Smyth DJ, Dorward DA, et al. A macrophage-pericyte axis directs tissue restoration via amphiregulin-induced transforming growth factor beta activation. Immunity. 2019;50:645-654.e6.PubMedPubMedCentralCrossRef
48.
McKee C, Sigala B, Soeda J, Mouralidarane A, Morgan M, Mazzoccoli G, et al. Amphiregulin activates human hepatic stellate cells and is upregulated in non alcoholic steatohepatitis. Sci Rep. 2015;5:8812.PubMedPubMedCentralCrossRef
49.
50.
Wilhelm A, Shepherd EL, Amatucci A, Munir M, Reynolds G, Humphreys E, et al. Interaction of TWEAK with Fn14 leads to the progression of fibrotic liver disease by directly modulating hepatic stellate cell proliferation: TWEAK as a regulator of liver fibrogenesis. J Pathol. 2016;239:109–21.PubMedPubMedCentralCrossRef
51.
Makino K, Makino T, Stawski L, Mantero JC, Lafyatis R, Simms R, et al. Blockade of PDGF receptors by crenolanib has therapeutic effect in patient fibroblasts and in preclinical models of systemic sclerosis. J Investig Dermatol. 2017;137:1671–81.PubMedCrossRef
Metadata
Title
Revealing potential anti-fibrotic mechanism of Ganxianfang formula based on RNA sequence
Authors
Zongyi Liu
Huanyu Xiang
Dejuan Xiang
Shuang Xiao
Hongyan Xiang
Jing Xiao
Hong Ren
Peng Hu
Huabao Liu
Mingli Peng
Publication date
01-12-2022
Publisher
BioMed Central
Published in
Chinese Medicine / Issue 1/2022
Electronic ISSN: 1749-8546
DOI
https://doi.org/10.1186/s13020-022-00579-7

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