Skip to main content
Top
Published in: Inflammation Research 6/2013

01-06-2013 | Original Research Paper

Effect of chlorogenic acid on LPS-induced proinflammatory signaling in hepatic stellate cells

Authors: Haitao Shi, Lei Dong, Xiaoyan Dang, Yaping Liu, Jiong Jiang, Yan Wang, Xiaolan Lu, Xiaoyan Guo

Published in: Inflammation Research | Issue 6/2013

Login to get access

Abstract

Objective and design

This study was aimed at investigating the effect of chlorogenic acid (CGA) on lipopolysaccharide (LPS)-induced proinflammatory signaling in hepatic stellate cells (HSCs).

Methods

An immortalized rat HSC line was cultured in vitro and treated with LPS in the absence or presence of CGA. Reactive oxygen species (ROS) production in the HSCs was monitored by flow cytometer using DCFH-DA. The protein expression levels of toll-like receptor 4 (TLR4), myeloid differentiation factor 88 (MyD88), nuclear factor-κB (NF-κB), and p-IκB-α were determined by Western blot. The mRNA expression levels of TLR4, MyD88, monocyte chemotactic protein 1(MCP-1), and interleukin 6 (IL-6) were detected by RT-PCR. The levels of MCP-1 and IL-6 in the culture supernatant of HSCs were measured by ELISA.

Results

CGA had no effect on expression of TLR4 and MyD88. However, the treatment of CGA can inhibit LPS-induced production of ROS in HSCs. Meanwhile, CGA can inhibit LPS-induced nuclear translocation of NF-κB and IκB-α phosphorylation in HSCs, as well as NAC (a ROS scavenger). The mRNA expression and the levels of MCP-1 and IL-6 in the culture supernatant of the HSCs in this study were elevated by LPS stimulation and inhibited by CGA treatment, as well as NAC and PDTC (a NF-κB inhibitor).

Conclusion

Our results indicate that CGA can efficiently inhibit LPS-induced proinflammatory responses in HSCs and the anti-inflammatory effect may be due to the inhibition of LPS/ROS/NF-κB signaling pathway.
Literature
1.
go back to reference Bataller R, Brenner DA. Liver fibrosis. J Clin Investig. 2005;115:209–18.PubMed Bataller R, Brenner DA. Liver fibrosis. J Clin Investig. 2005;115:209–18.PubMed
2.
go back to reference Friedman SL. Evolving challenges in hepatic fibrosis. Nat Rev Gastroenterol and Hepatol. 2010;7:425–36.CrossRef Friedman SL. Evolving challenges in hepatic fibrosis. Nat Rev Gastroenterol and Hepatol. 2010;7:425–36.CrossRef
3.
go back to reference Fallowfield JA. Therapeutic targets in liver fibrosis. Am J Physiol-Gastrointest Liver Physiol. 2011;300:G709–15.PubMedCrossRef Fallowfield JA. Therapeutic targets in liver fibrosis. Am J Physiol-Gastrointest Liver Physiol. 2011;300:G709–15.PubMedCrossRef
5.
go back to reference Saile B, Ramadori G. Inflammation, damage repair and liver fibrosis—role of cytokines and different cell types. Z Gastroent. 2007;45:77–86.CrossRef Saile B, Ramadori G. Inflammation, damage repair and liver fibrosis—role of cytokines and different cell types. Z Gastroent. 2007;45:77–86.CrossRef
6.
go back to reference Pinzani M, Macias-Barragan J. Update on the pathophysiology of liver fibrosis. Expert Rev Gastroenterol Hepatol. 2010;4:459–72.PubMedCrossRef Pinzani M, Macias-Barragan J. Update on the pathophysiology of liver fibrosis. Expert Rev Gastroenterol Hepatol. 2010;4:459–72.PubMedCrossRef
7.
go back to reference O’Neill LA. Therapeutic targeting of Toll-like receptors for inflammatory and infectious diseases. Curr Opin Pharmacol. 2003;3:396–403.PubMedCrossRef O’Neill LA. Therapeutic targeting of Toll-like receptors for inflammatory and infectious diseases. Curr Opin Pharmacol. 2003;3:396–403.PubMedCrossRef
8.
go back to reference Zhu JK, Mohan C. Toll-like receptor signaling pathways-therapeutic opportunities. Mediat Inflamm. 2010. Art ID 781235. Zhu JK, Mohan C. Toll-like receptor signaling pathways-therapeutic opportunities. Mediat Inflamm. 2010. Art ID 781235.
9.
go back to reference Aoyama T, Paik YH, Seki E. Toll-like receptor signaling and liver fibrosis. Gastroenterol Res Pract. 2010. Art ID 192543. Aoyama T, Paik YH, Seki E. Toll-like receptor signaling and liver fibrosis. Gastroenterol Res Pract. 2010. Art ID 192543.
10.
go back to reference Lin RS, Lee FY, Lee SD, Tsai YT, Lin HC, Lu RH, et al. Endotoxemia in patients with chronic liver-diseases—relationship to severity of liver diseases, presence of esophageal varices, and hyperdynamic circulation. J Hepatol. 1995;22:165–72.PubMedCrossRef Lin RS, Lee FY, Lee SD, Tsai YT, Lin HC, Lu RH, et al. Endotoxemia in patients with chronic liver-diseases—relationship to severity of liver diseases, presence of esophageal varices, and hyperdynamic circulation. J Hepatol. 1995;22:165–72.PubMedCrossRef
11.
go back to reference Parlesak A, Schafer C, Schutz T, Bode JC, Bode C. Increased intestinal permeability to macromolecules and endotoxemia in patients with chronic alcohol abuse in different stages of alcohol-induced liver disease. J Hepatol. 2000;32:742–7.PubMedCrossRef Parlesak A, Schafer C, Schutz T, Bode JC, Bode C. Increased intestinal permeability to macromolecules and endotoxemia in patients with chronic alcohol abuse in different stages of alcohol-induced liver disease. J Hepatol. 2000;32:742–7.PubMedCrossRef
12.
go back to reference Choi Y, Jeon WK, Hwang SJ, Kim BI, Sohn CI, Park DI, et al. The Role of the Gut Barrier Function in the Pathophysiology of Viral Liver Cirrhosis. Hepato-Gastroenterol. 2011;58:1244–7. Choi Y, Jeon WK, Hwang SJ, Kim BI, Sohn CI, Park DI, et al. The Role of the Gut Barrier Function in the Pathophysiology of Viral Liver Cirrhosis. Hepato-Gastroenterol. 2011;58:1244–7.
13.
go back to reference Seki E, Brenner DA. Toll-like receptors and adaptor molecules in liver disease: update. Hepatology. 2008;48:322–35.PubMedCrossRef Seki E, Brenner DA. Toll-like receptors and adaptor molecules in liver disease: update. Hepatology. 2008;48:322–35.PubMedCrossRef
14.
go back to reference Brun P, Castagliuolo I, Pinzani M, Palu G, Martines D. Exposure to bacterial cell wall products triggers an inflammatory phenotype in hepatic stellate cells. Am J Physiol-Gastroint Liver Physiol. 2005;289:G571–8.CrossRef Brun P, Castagliuolo I, Pinzani M, Palu G, Martines D. Exposure to bacterial cell wall products triggers an inflammatory phenotype in hepatic stellate cells. Am J Physiol-Gastroint Liver Physiol. 2005;289:G571–8.CrossRef
15.
go back to reference Paik YH, Schwabe RF, Bataller R, Russo MP, Jobin C, Brenner DA. Toll-like receptor 4 mediates inflammatory signaling by bacterial lipopolysaccharide in human hepatic stellate cells. Hepatology. 2003;37:1043–55.PubMedCrossRef Paik YH, Schwabe RF, Bataller R, Russo MP, Jobin C, Brenner DA. Toll-like receptor 4 mediates inflammatory signaling by bacterial lipopolysaccharide in human hepatic stellate cells. Hepatology. 2003;37:1043–55.PubMedCrossRef
16.
go back to reference Thirunavukkarasu C, Watkins SC, Gandhi CR. Mechanisms of endotoxin-induced NO, IL-6, and TNF-alpha production in activated rat hepatic stellate cells: role of p38 MAPK. Hepatol. 2006;44:389–98.CrossRef Thirunavukkarasu C, Watkins SC, Gandhi CR. Mechanisms of endotoxin-induced NO, IL-6, and TNF-alpha production in activated rat hepatic stellate cells: role of p38 MAPK. Hepatol. 2006;44:389–98.CrossRef
17.
go back to reference Seki E, De Minicis S, Osterreicher CH, Kluwe J, Osawa Y, Brenner DA, et al. TLR4 enhances TGF-beta signaling and hepatic fibrosis. Nat Med. 2007;13:1324–32.PubMedCrossRef Seki E, De Minicis S, Osterreicher CH, Kluwe J, Osawa Y, Brenner DA, et al. TLR4 enhances TGF-beta signaling and hepatic fibrosis. Nat Med. 2007;13:1324–32.PubMedCrossRef
18.
go back to reference Feng RT, Lu YJ, Bowman LL, Qian Y, Castranova V, Ding M. Inhibition of activator protein-1, NF-kappa B, and MAPKs and induction of phase 2 detoxifying enzyme activity by chlorogenic acid. J Biol Chem. 2005;280:27888–95.PubMedCrossRef Feng RT, Lu YJ, Bowman LL, Qian Y, Castranova V, Ding M. Inhibition of activator protein-1, NF-kappa B, and MAPKs and induction of phase 2 detoxifying enzyme activity by chlorogenic acid. J Biol Chem. 2005;280:27888–95.PubMedCrossRef
19.
go back to reference Zhang X, Huang H, Yang TT, Ye Y, Shan JH, Yin ZM, et al. Chlorogenic acid protects mice against lipopolysaccharide-induced acute lung injury. Inj Int J Care Inj. 2010;41:746–52. Zhang X, Huang H, Yang TT, Ye Y, Shan JH, Yin ZM, et al. Chlorogenic acid protects mice against lipopolysaccharide-induced acute lung injury. Inj Int J Care Inj. 2010;41:746–52.
20.
go back to reference Kono Y, Kobayashi K, Tagawa S, Adachi K, Ueda A, Sawa Y, et al. Antioxidant activity of polyphenolics in diets Rate constants of reactions of chlorogenic acid and caffeic acid with reactive species of oxygen and nitrogen. Biochim Biophys Acta. 1997;1335:335–42.PubMedCrossRef Kono Y, Kobayashi K, Tagawa S, Adachi K, Ueda A, Sawa Y, et al. Antioxidant activity of polyphenolics in diets Rate constants of reactions of chlorogenic acid and caffeic acid with reactive species of oxygen and nitrogen. Biochim Biophys Acta. 1997;1335:335–42.PubMedCrossRef
21.
go back to reference Shi HY, Dong L, Bai YH, Zhao JH, Zhang Y, Zhang L. Chlorogenic acid against carbon tetrachloride-induced liver fibrosis in rats. Eur J Pharmacol. 2009;623:119–24.PubMedCrossRef Shi HY, Dong L, Bai YH, Zhao JH, Zhang Y, Zhang L. Chlorogenic acid against carbon tetrachloride-induced liver fibrosis in rats. Eur J Pharmacol. 2009;623:119–24.PubMedCrossRef
22.
go back to reference Xu YX, Chen JW, Yu XA, Tao WW, Jiang FR, Yin ZM, et al. Protective effects of chlorogenic acid on acute hepatotoxicity induced by lipopolysaccharide in mice. Inflamm Res. 2010;59:871–7.PubMedCrossRef Xu YX, Chen JW, Yu XA, Tao WW, Jiang FR, Yin ZM, et al. Protective effects of chlorogenic acid on acute hepatotoxicity induced by lipopolysaccharide in mice. Inflamm Res. 2010;59:871–7.PubMedCrossRef
23.
go back to reference Shan JH, Fu J, Zhao ZH, Kong XQ, Huang H, Luo L, et al. Chlorogenic acid inhibits lipopolysaccharide-induced cyclooxygenase-2 expression in RAW264.7 cells through suppressing NF-kappa B and JNK/AP-1 activation. Int Immunopharmacol. 2009;9:1042–8.PubMedCrossRef Shan JH, Fu J, Zhao ZH, Kong XQ, Huang H, Luo L, et al. Chlorogenic acid inhibits lipopolysaccharide-induced cyclooxygenase-2 expression in RAW264.7 cells through suppressing NF-kappa B and JNK/AP-1 activation. Int Immunopharmacol. 2009;9:1042–8.PubMedCrossRef
24.
go back to reference Wang BB, Cheng JY, Gao HH, Zhang Y, Chen ZN, Bian HJ. Hepatic stellate cells in inflammation-fibrosis-carcinoma axis. Anat Rec. 2010;293:1492–6.CrossRef Wang BB, Cheng JY, Gao HH, Zhang Y, Chen ZN, Bian HJ. Hepatic stellate cells in inflammation-fibrosis-carcinoma axis. Anat Rec. 2010;293:1492–6.CrossRef
25.
go back to reference Vogel S, Piantedosi R, Frank J, Lalazar A, Rockey DC, Friedman SL, et al. An immortalized rat liver stellate cell line (HSC-T6): a new cell model for the study of retinoid metabolism in vitro. J Lipid Res. 2000;41:882–93.PubMed Vogel S, Piantedosi R, Frank J, Lalazar A, Rockey DC, Friedman SL, et al. An immortalized rat liver stellate cell line (HSC-T6): a new cell model for the study of retinoid metabolism in vitro. J Lipid Res. 2000;41:882–93.PubMed
26.
go back to reference Lebel CP, Ischiropoulos H, Bondy SC. Evaluation of the probe 2′, 7′-dichlorofluorescin as an indicator of reactive oxygen species formation and oxidative stress. Chem Res Toxicol. 1992;5:227–31.PubMedCrossRef Lebel CP, Ischiropoulos H, Bondy SC. Evaluation of the probe 2′, 7′-dichlorofluorescin as an indicator of reactive oxygen species formation and oxidative stress. Chem Res Toxicol. 1992;5:227–31.PubMedCrossRef
27.
go back to reference Choi HK, Pokharel YR, Lim SC, Han HK, Ryu CS, Kim SK, et al. Inhibition of liver fibrosis by solubilized coenzyme Q10: Role of Nrf2 activation in inhibiting transforming growth factor-beta 1 expression. Toxicol Appl Pharmacol. 2009;240:377–84.PubMedCrossRef Choi HK, Pokharel YR, Lim SC, Han HK, Ryu CS, Kim SK, et al. Inhibition of liver fibrosis by solubilized coenzyme Q10: Role of Nrf2 activation in inhibiting transforming growth factor-beta 1 expression. Toxicol Appl Pharmacol. 2009;240:377–84.PubMedCrossRef
28.
go back to reference Gill R, Tsung A, Billiar T. Linking oxidative stress to inflammation: Toll-like receptors. Free Radical Biol Med. 2010;48:1121–32.CrossRef Gill R, Tsung A, Billiar T. Linking oxidative stress to inflammation: Toll-like receptors. Free Radical Biol Med. 2010;48:1121–32.CrossRef
29.
go back to reference Wu J, Meng ZJ, Jiang M, Zhang EJ, Trippler M, Broering R, et al. Toll-like receptor-induced innate immune responses in non-parenchymal liver cells are cell type-specific. Immunology. 2010;129:363–74.PubMedCrossRef Wu J, Meng ZJ, Jiang M, Zhang EJ, Trippler M, Broering R, et al. Toll-like receptor-induced innate immune responses in non-parenchymal liver cells are cell type-specific. Immunology. 2010;129:363–74.PubMedCrossRef
30.
31.
go back to reference DiDonato JA, Hayakawa M, Rothwarf DM, Zandi E, Karin M. A cytokine-responsive IkappaB kinase that activates the transcription factor NF-kappaB. Nature. 1997;388:548–54.PubMedCrossRef DiDonato JA, Hayakawa M, Rothwarf DM, Zandi E, Karin M. A cytokine-responsive IkappaB kinase that activates the transcription factor NF-kappaB. Nature. 1997;388:548–54.PubMedCrossRef
32.
go back to reference Zandi E, Rothwarf DM, Delhase M, Hayakawa M, Karin M. The I kappa B kinase complex (IKK) contains two kinase subunits, IKK alpha and IKK beta, necessary for I kappa B phosphorylation and NF-kappa B activation. Cell. 1997;91:243–52.PubMedCrossRef Zandi E, Rothwarf DM, Delhase M, Hayakawa M, Karin M. The I kappa B kinase complex (IKK) contains two kinase subunits, IKK alpha and IKK beta, necessary for I kappa B phosphorylation and NF-kappa B activation. Cell. 1997;91:243–52.PubMedCrossRef
33.
go back to reference Marra F, DeFranco R, Grappone C, Milani S, Pastacaldi S, Pinzani M, et al. Increased expression of monocyte chemotactic protein-1 during active hepatic fibrogenesis: correlation with monocyte infiltration. Am J Pathol. 1998;152:423–30.PubMed Marra F, DeFranco R, Grappone C, Milani S, Pastacaldi S, Pinzani M, et al. Increased expression of monocyte chemotactic protein-1 during active hepatic fibrogenesis: correlation with monocyte infiltration. Am J Pathol. 1998;152:423–30.PubMed
34.
go back to reference Wasmuth HE, Tacke F, Trautwein C. Chemokines in liver inflammation and fibrosis. Semin Liver Dis. 2010;30:215–25.PubMedCrossRef Wasmuth HE, Tacke F, Trautwein C. Chemokines in liver inflammation and fibrosis. Semin Liver Dis. 2010;30:215–25.PubMedCrossRef
35.
go back to reference Seki E, De Minicis S, Inokuchi S, Taura K, Miyai K, Van Rooijen N, et al. CCR2 Promotes Hepatic Fibrosis in Mice. Hepatology. 2009;50:185–97.PubMedCrossRef Seki E, De Minicis S, Inokuchi S, Taura K, Miyai K, Van Rooijen N, et al. CCR2 Promotes Hepatic Fibrosis in Mice. Hepatology. 2009;50:185–97.PubMedCrossRef
36.
go back to reference Marra F, Romanelli RG, Giannini C, Failli P, Pastacaldi S, Arrighi MC, et al. Monocyte chemotactic protein-1 as a chemoattractant for human hepatic stellate cells. Hepatology. 1999;29:140–8.PubMedCrossRef Marra F, Romanelli RG, Giannini C, Failli P, Pastacaldi S, Arrighi MC, et al. Monocyte chemotactic protein-1 as a chemoattractant for human hepatic stellate cells. Hepatology. 1999;29:140–8.PubMedCrossRef
37.
go back to reference Tsuruta S, Nakamuta M, Enjoji M, Kotoh K, Hiasa K, Egashira K, et al. Anti-monocyte chemoattractant protein-1 gene therapy prevents dimethylnitrosamine-induced hepatic fibrosis in rats. Int J Mol Med. 2004;14:837–42.PubMed Tsuruta S, Nakamuta M, Enjoji M, Kotoh K, Hiasa K, Egashira K, et al. Anti-monocyte chemoattractant protein-1 gene therapy prevents dimethylnitrosamine-induced hepatic fibrosis in rats. Int J Mol Med. 2004;14:837–42.PubMed
38.
go back to reference Kayano K, Okita K. Does IL-6 regulate liver fibrosis/cirrhosis directly and indirectly? J Gastroenterol. 2000;35:250–1.PubMedCrossRef Kayano K, Okita K. Does IL-6 regulate liver fibrosis/cirrhosis directly and indirectly? J Gastroenterol. 2000;35:250–1.PubMedCrossRef
39.
go back to reference Choi I, Kang HS, Yang Y, Pyun KH. IL-6 induces hepatic inflammation and collagen-synthesis in vivo. Clin Exp Immunol. 1994;95:530–5.PubMedCrossRef Choi I, Kang HS, Yang Y, Pyun KH. IL-6 induces hepatic inflammation and collagen-synthesis in vivo. Clin Exp Immunol. 1994;95:530–5.PubMedCrossRef
40.
go back to reference Asehnoune K, Strassheim D, Mitra S, Kim JY, Abraham E. Involvement of reactive oxygen species in Toll-like receptor 4-dependent activation of NF-kappaB. J Immunol. 2004;172:2522–9.PubMed Asehnoune K, Strassheim D, Mitra S, Kim JY, Abraham E. Involvement of reactive oxygen species in Toll-like receptor 4-dependent activation of NF-kappaB. J Immunol. 2004;172:2522–9.PubMed
41.
go back to reference Park HS, Jung HY, Park EY, Kim J, Lee WJ, Bae YS. Cutting edge: direct interaction of TLR4 with NAD(P)H oxidase 4 isozyme is essential for lipopolysaccharide-induced production of reactive oxygen species and activation of NF-kappaB. J Immunol. 2004;173:3589–93.PubMed Park HS, Jung HY, Park EY, Kim J, Lee WJ, Bae YS. Cutting edge: direct interaction of TLR4 with NAD(P)H oxidase 4 isozyme is essential for lipopolysaccharide-induced production of reactive oxygen species and activation of NF-kappaB. J Immunol. 2004;173:3589–93.PubMed
Metadata
Title
Effect of chlorogenic acid on LPS-induced proinflammatory signaling in hepatic stellate cells
Authors
Haitao Shi
Lei Dong
Xiaoyan Dang
Yaping Liu
Jiong Jiang
Yan Wang
Xiaolan Lu
Xiaoyan Guo
Publication date
01-06-2013
Publisher
SP Birkhäuser Verlag Basel
Published in
Inflammation Research / Issue 6/2013
Print ISSN: 1023-3830
Electronic ISSN: 1420-908X
DOI
https://doi.org/10.1007/s00011-013-0610-7

Other articles of this Issue 6/2013

Inflammation Research 6/2013 Go to the issue
Live Webinar | 27-06-2024 | 18:00 (CEST)

Keynote webinar | Spotlight on medication adherence

Live: Thursday 27th June 2024, 18:00-19:30 (CEST)

WHO estimates that half of all patients worldwide are non-adherent to their prescribed medication. The consequences of poor adherence can be catastrophic, on both the individual and population level.

Join our expert panel to discover why you need to understand the drivers of non-adherence in your patients, and how you can optimize medication adherence in your clinics to drastically improve patient outcomes.

Prof. Kevin Dolgin
Prof. Florian Limbourg
Prof. Anoop Chauhan
Developed by: Springer Medicine
Obesity Clinical Trial Summary

At a glance: The STEP trials

A round-up of the STEP phase 3 clinical trials evaluating semaglutide for weight loss in people with overweight or obesity.

Developed by: Springer Medicine