Effect of interleukin-6 neutralization on CYP3A11 and metallothionein-1/2 expressions in arthritic mouse liver

doi:10.1016/j.ejphar.2006.11.072

European Journal of Pharmacology

Volume 558, Issues 1–3, 8 March 2007, Pages 199-207

https://doi.org/10.1016/j.ejphar.2006.11.072 Get rights and content

Abstract

Rheumatoid arthritis is characterized by chronic inflammation of the synovial tissue. We examined the effect of interleukin (IL)-6 neutralization on the expression of cytochrome P450 or metallothionein-1/2 (metallothionein) during chronic phase inflammatory disease using rheumatoid arthritis model mice, human T-cell leukemia virus type I (HTLV-I) transgenic mice. Serum IL-6 concentrations of arthritis-developed HTLV-I transgenic mice were 129.9 ± 26.1 pg/ml. Moreover, signal transducer and activator of transcription (STAT) 1/3 phosphorylations was observed in arthritic HTLV-I transgenic mouse livers. CYP3A11 mRNA was more strongly reduced by the development of arthritis in HTLV-I transgenic mouse livers as compared with CYP2C29 or CYP2E1 mRNAs. CYP3A protein and testosterone 6β-hydroxylation activity also changed in a similar manner to the corresponding CYP3A11 mRNA level. On the other hand, metallothionein mRNA was significantly induced as compared with that of wild-type or non-arthritic mice. CYP3A suppression and metallothionein mRNA overexpression activity seen in the developed arthritic mice returned to the gene conditions of the non-arthritic HTLV-I transgenic mice by IL-6 antibody at 48 h after treatment. The present study has revealed that CYP3A11 and metallothionein expressions are affected by the release of IL-6 by arthritis and its systemic circulation, and neutralization of IL-6 recovered from the down-regulation of CYP3A11 mRNA and the induction of metallothionein mRNA in arthritic HTLV-I transgenic mice.

Introduction

Cytochrome P450 superfamily monooxygenases are important for the metabolism of variety of an endogenous and exogenous compounds. The regulation of cytochrome P450 genes and the variation of cytochrome P450 activities can be affected by a number of factors, such as xenobiotics and diseases. Suppression of the drug-metabolizing capability results in various adverse effects and/or elevation of blood drug concentrations. It has been known that during infections or inflammation, the hepatic drug-metabolizing capability is reduced in experimental animals and humans (Iber et al., 1999, Morgan, 1997). This phenomenon suggests that infections and inflammation down-regulate cytochrome P450 such as CYP3A, cytochrome P450 gene expressions and their protein enzymatic activities (Li-Masters and Morgan, 2002, Sewer et al., 1998). Evidence has revealed that inflammatory cytokines, such as interleukin (IL)-1β, IL-6 and tumor necrosis factor (TNF) α, play important roles in cytochrome P450 mRNA regulation (Morgan, 1997). Down-regulation of cytochrome P450 families results in changes of pharmacokinetics, metabolism, distribution and excretion of endogenous substances, exogenous drugs and chemicals. Many down-regulation studies concerned with cytochrome P450 families have focused on the acute phase response to proinflammatory substances such as lipopolysaccharide (LPS), turpentine, or Bacillus Calmette-Guerin administration (Ashino et al., 2004, Siewert et al., 2000). On the other hand, the changes in cytochrome P450 families under chronic inflammatory conditions have not yet been elucidated.

Rheumatoid arthritis is one of the most common autoimmune diseases (Firestein and Zvaifler, 1992). It is clearly shown that IL-1 and TNFα play important roles in the development of rheumatoid arthritis (Dayer, 2002, Taylor, 2003). Furthermore, rheumatoid arthritis pathogenesis itself enhances production of not only IL-1 and TNFα but also other cytokines such as IL-6 (Feldmann et al., 1996, Horai et al., 2000). Human T-cell leukemia virus type I (HTLV-I) is known as the causative insult of adult T-cell leukemia (Sugamura and Hinuma, 1993, Yoshida, 1993). Nishioka et al. (1989) firstly reported a patient with chronic inflammatory arthropathy infected with HTLV-I. Subsequently, Iwakura et al. (1991) found that HTLV-I could induce arthritis in mice. The HTLV-I transgenic mice produced by Iwakura et al. (1991) have been shown to develop chronic inflammatory polyarthropathy with a high incidence. The histopathology of HTLV-I transgenic mice is very similar to that of rheumatoid arthritis in humans, showing marked synovial and periarticular inflammation with articular erosion caused by invasion of granulation tissues (Yamamoto et al., 1993). It seems that the HTLV-I transgenic mouse is likely to be the best animal model for rheumatoid arthritis.

Metallothionein-1/2 (metallothionein) is a small, cysteine-rich, metal-binding protein. Metallothionein is capable of not only reducing metal toxicity but also scavenging free radicals or regulating metal-metabolism. Metallothionein synthesis is induced by various stimuli such as heavy metals, oxidative stress, cytokines and inflammatory diseases (Oguro and Yoshida, 2004). Metallothionein induction by LPS has been shown to be regulated through IL-6 proofed by IL-6 gene deficient mice. A signal transducer and activator of transcription (STAT) 1/STAT3 binding site exists in the promoter region of the metallothionein gene suggesting that STAT1/3 mediated signal pathway is a dominant signal transduction for LPS-mediated metallothionein induction (Lee et al., 1999).

Rheumatoid arthritis is characterized by not only chronic inflammation of the multiple joints but also systemic inflammatory responses. In particular, the liver expresses various enzymes such as drug metabolizing enzymes and is subject to affect inflammatory diseases and/or oxidative stress. Furthermore, studies on cytochrome P450 and metallothionein gene expressions in the livers by IL-6 neutralization are scarce and remain to be explored though IL-6 neutralizing antibody is used to treat rheumatoid arthritis. Considering the aforementioned findings, we investigated the relationship between changes in cytochrome P450 and metallothionein expressions and IL-6 neutralization in chronic inflammation disease model mice, HTLV-I transgenic mice.

Section snippets

Reagents

Deoxycytidine 5′-[α-³²P]-triphosphate (3000 Ci/mmol) and adenosine 5′-[γ-³²P]-triphosphate (5000 Ci/mmol) were obtained from Japan Isotope Association (Tokyo, Japan). Complete protease inhibitor cocktail was obtained from Roche Diagnostics (Mannheim, Germany). Anti-rat polyclonal CYP3A2 antibodies were obtained from Daiichi Pure Chemicals Co., Ltd. (Tokyo, Japan). Monoclonal antibody against STAT1 (C terminus) and STAT3 was obtained from BD Biosciences Transduction Laboratories (Lexington, KY).

Arthritic mouse and change of serum inflammatory cytokine concentrations by development of rheumatoid arthritis

Fig. 1-A shows the features of non-arthritic mouse and arthritic mouse, respectively. The posterior limb joints of the arthritic mouse were swollen as compared with those of the non-arthritic mouse. Moreover, some mice used in this study showed swelling of anterior limb joints in addition to posterior joints. HTLV-I transgenic mice start to develop arthritis spontaneously at 4 weeks of age, and 60% and 80% of the mice are affected at 3 months and 6 months of age, respectively. The ages of

Discussion

Mechanistic studies on the development of rheumatoid arthritis have been carried out by many researchers. Recently, human-type anti-inflammatory cytokine antibodies, IL-1β, IL-6, TNFα and their receptors, have been employed and developed for rheumatoid arthritis therapy resulting in effective treatment for rheumatoid arthritis. However, the effects of cytokine targeted therapies on other organs remain to be explored. In this study, we focused on the effect of IL-6 neutralization and

Acknowledgments

We thank J. Sugiuchi and H. Tomiyama for their technical assistance. We also thank Dr. A. Naganuma (Laboratory of Molecular and Biochemical Toxicology, Tohoku University, Sendai, Japan) and Dr. K. Nose (Department of Microbiology, Showa University) for kindly providing cDNA probes of mouse metallothionein-1 and rat GAPDH, respectively. A part of this study was supported by a Grant-in-Aid from the Ministry of Education, Science and Culture, Japan.

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Effect of interleukin-6 neutralization on CYP3A11 and metallothionein-1/2 expressions in arthritic mouse liver

Abstract

Introduction

Section snippets

Reagents

Arthritic mouse and change of serum inflammatory cytokine concentrations by development of rheumatoid arthritis

Discussion

Acknowledgments

Biochem. Biophys. Res. Commun.

Anal. Biochem.

Anal. Biochem.

J. Card. Fail.

Biochem. Pharmacol.

J. Biol. Chem.

Lancet

J. Biol. Chem.

Hepatology

Anal. Biochem.

Arch. Biochem. Biophys.

Drug Metab. Pharmacokinet.

Trends Microbiol.

Involvement of interleukin-6 and tumor necrosis factor alpha in CYP3A11 and 2C29 down-regulation bacillus Calmette-Guerin and lipopolysaccharide in mouse liver

Drug Metab. Dispos.

Metallothionein: the multipurpose protein

Cell. Mol. Life Sci.

Interleukin 1 or tumor necrosis factor-alpha: which is the real target in rheumatoid arthritis?

J. Rheumatol., Suppl.

Role of cytokines in rheumatoid arthritis

Annu. Rev. Immunol.