Suppression of interleukin-6-induced C-reactive protein expression by FXR agonists

doi:10.1016/j.bbrc.2008.12.117

Biochemical and Biophysical Research Communications

Volume 379, Issue 2, 6 February 2009, Pages 476-479

https://doi.org/10.1016/j.bbrc.2008.12.117 Get rights and content

Abstract

C-reactive protein (CRP), a human acute-phase protein, is a risk factor for future cardiovascular events and exerts direct pro-inflammatory and pro-atherogenic properties. The farnesoid X receptor (FXR), a member of the nuclear hormone receptor superfamily, plays an essential role in the regulation of enterohepatic circulation and lipid homeostasis. In this study, we report that two synthetic FXR agonists, WAY-362450 and GW4064, suppressed interleukin-6-induced CRP expression in human Hep3B hepatoma cells. Knockdown of FXR by short interfering RNA attenuated the inhibitory effect of the FXR agonists and also increased the ability of interleukin-6 to induce CRP production. Furthermore, treatment of wild type C57BL/6 mice with the FXR agonist, WAY-362450, attenuated lipopolysaccharide-induced serum amyloid P component and serum amyloid A3 mRNA levels in the liver, whereas no effect was observed in FXR knockout mice. These data provide new evidence for direct anti-inflammatory properties of FXR.

Section snippets

Materials and methods

Cell culture, treatment and siRNA transfection. Human hepatoma Hep3B cells (ATCC, Rockville, MD) were cultured in MEM supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin. For treatment, cells were washed, incubated in serum-free medium, and stimulated with 50 ng/mL IL-6 in the presence of DMSO, 1 μmol/L WAY-362450 or 1 μmol/L GW4064. After 24 h of treatment, the supernatants were collected for CRP measurement with a commercial ELISA kit from Meso scale (K151EPC-3), and the

FXR agonists suppress IL-6-induced CRP expression in Hep3B cells

Since human hepatoma cell line Hep3B is a superior model to investigate CRP acute phase response in human hepatocytes [12], [24], [25], we chose the Hep3B cells to investigate whether FXR agonists modulate IL-6-induced CRP production. Hep3B cells were stimulated with 50 ng/mL IL-6 in the presence or absence of WAY-362450 or GW4064. After 24 h of treatment, CRP levels were determined in the cell supernatants by ELISA. No CRP protein was detected in unstimulated Hep3B cell supernatants, whereas

Discussion

In FXR-deficient mice, hepatic mRNA levels of inflammation markers, including interferon γ, tumor necrosis factor-α and interleukin-1β, were significantly higher than those in wild-type controls [26], [27], [28]. Moreover, LPS administration induces hepatic acute phase response, and decreases the mRNA levels of FXR and its target genes in C57BL/6 mice [29]. However, there are no reports that establish the direct role of FXR in hepatic inflammation and acute phase response. In this report, we

Conflict of interest

Songwen Zhang, Qiangyuan Liu, Juan Wang and Douglas C. Harnish are employees of Wyeth.

Acknowledgments

We thank the Wyeth Bioresource Department for support of in vivo studies, Evans M.J. for discussions and assistance with the experiments.

References (30)

R. Kleemann et al.
Fibrates down-regulate IL-1-stimulated C-reactive protein gene expression in hepatocytes by reducing nuclear p50-NFkappa B-C/EBP-beta complex formation
Blood
(2003)
B.M. Forman et al.
Identification of a nuclear receptor that is activated by farnesol metabolites
Cell
(1995)
Y. Zhang et al.
FXR signaling in metabolic disease
FEBS Lett.
(2008)
H. Wang et al.
Endogenous bile acids are ligands for the nuclear receptor FXR/BAR
Mol. Cell
(1999)
M.S. Kim et al.
Repression of farnesoid X receptor during the acute phase response
J. Biol. Chem.
(2003)
P.M. Ridker
Clinical application of C-reactive protein for cardiovascular disease detection and prevention
Circulation
(2003)
F. Haverkate et al.
Production of C-reactive protein and risk of coronary events in stable and unstable angina. European Concerted Action on Thrombosis and Disabilities Angina Pectoris Study Group
Lancet
(1997)
V. Pasceri et al.
Direct proinflammatory effect of C-reactive protein on human endothelial cells
Circulation
(2000)
M. Torzewski et al.
C-reactive protein in the arterial intima: role of C-reactive protein receptor-dependent monocyte recruitment in atherogenesis
Arterioscler. Thromb. Vasc. Biol.
(2000)
S.K. Venugopal et al.
Effect of C-reactive protein on vascular cells: evidence for a proinflammatory, proatherogenic role
Curr. Opin. Nephrol. Hypertens.
(2005)

C.H. Wang et al.

C-reactive protein upregulates angiotensin type 1 receptors in vascular smooth muscle

Circulation

(2003)

T.P. Zwaka et al.

C-reactive protein-mediated low density lipoprotein uptake by macrophages: implications for atherosclerosis

Circulation

(2001)

M.K. Ganapathi et al.

Effect of combinations of cytokines and hormones on synthesis of serum amyloid A and C-reactive protein in Hep 3B cells

J. Immunol.

(1991)

B. Desvergne et al.

Peroxisome proliferator-activated receptors: nuclear control of metabolism

Endocr. Rev.

(1999)

B. Staels et al.

Activation of human aortic smooth-muscle cells is inhibited by PPARalpha but not by PPARgamma activators

Nature

(1998)

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Suppression of interleukin-6-induced C-reactive protein expression by FXR agonists

Abstract

Section snippets

Materials and methods

FXR agonists suppress IL-6-induced CRP expression in Hep3B cells

Discussion

Conflict of interest

Acknowledgments

Blood

Cell

FEBS Lett.

Mol. Cell

J. Biol. Chem.

Clinical application of C-reactive protein for cardiovascular disease detection and prevention

Circulation

Production of C-reactive protein and risk of coronary events in stable and unstable angina. European Concerted Action on Thrombosis and Disabilities Angina Pectoris Study Group

Lancet

Direct proinflammatory effect of C-reactive protein on human endothelial cells

Circulation

C-reactive protein in the arterial intima: role of C-reactive protein receptor-dependent monocyte recruitment in atherogenesis

Arterioscler. Thromb. Vasc. Biol.

Effect of C-reactive protein on vascular cells: evidence for a proinflammatory, proatherogenic role

Curr. Opin. Nephrol. Hypertens.

C-reactive protein upregulates angiotensin type 1 receptors in vascular smooth muscle

Circulation

C-reactive protein-mediated low density lipoprotein uptake by macrophages: implications for atherosclerosis

Circulation

Effect of combinations of cytokines and hormones on synthesis of serum amyloid A and C-reactive protein in Hep 3B cells

J. Immunol.

Peroxisome proliferator-activated receptors: nuclear control of metabolism

Endocr. Rev.

Activation of human aortic smooth-muscle cells is inhibited by PPARalpha but not by PPARgamma activators

Nature