PENTOXIFYLLINE DIMINISHED ACETALDEHYDE-INDUCED COLLAGEN PRODUCTION IN HEPATIC STELLATE CELLS BY DECREASING INTERLEUKIN-6 EXPRESSION

doi:10.1016/S1043661802002025

Pharmacological Research

Volume 46, Issue 5, November 2002, Pages 435-443

https://doi.org/10.1016/S1043661802002025 Get rights and content

Abstract

The effect of pentoxifylline (PTX), a methylxanthine derivative, on collagen induction and secretion and on the production of mRNA of two fibrogenic cytokines: interleukin-6 and transforming growth factor-β₁ (IL-6 and TGF-β₁) in a rat hepatic stellate cell line (CFSC-2G) exposed to acetaldehyde was studied. CFSC-2G cells were treated with 175 μM acetaldehyde for 24 h. The cells were then exposed to a medium containing 200 μM PTX. Collagen secretion, increased 2.6 times in acetaldehyde treated cells. Cells exposed to acetaldehyde and treated with PTX diminished collagen secretion to control values and decreased α₁(I) collagen mRNA by 15%. Reverse transcriptase-polymerase chain reaction (RT-PCR) assays of TGF-β₁ mRNA showed no variation in different experimental conditions. However, PTX induced a decrease of 32% in IL-6 mRNA in acetaldehyde-treated cells. CFSC-2G cells treated with anti-IL-6 monoclonal antibody, 15 min before acetaldehyde was added, did not present an increase in α₁(I) collagen mRNA. These results show that PTX inhibits the expression of α₁(I) collagen via the inhibition of IL-6 in acetaldehyde treated cells. The effect herein reported on IL-6 and α₁(I) collagen mRNA adds to the previously described effect of PTX, which could be useful in the fibrogenic process induced by acetaldehyde.

Section snippets

INTRODUCTION

Liver fibrosis is the common consequence of different liver diseases characterized by chronic liver tissue damage. This process is a consequence of chronic activation of hepatic stellate cells (HSC), leading to cell proliferation and increased deposition of extracellular matrix components [1]. Data from different laboratories suggest that ethanol first metabolite acetaldehyde is highly fibrogenic per se. Acetaldehyde enhances the synthesis of α₁(I) collagen in hepatic cultured stellate cells

Cell culture

All the experiments were performed using the rat hepatic stellate cell line CFSC-2G, which was obtained and kindly donated to us by Dr Rojkind (Greenwel et al. [4]). Cells were cultured in a minimal essential medium (MEM), (Gibco) supplemented with 10% bovine fetal serum (Hyclone Laboratories Inc., Logan, UT, USA), nonessential amino acids, penicillin (100 U ml⁻¹) and streptomycin (100 mg ml⁻¹). Cells were grown at 37 °C in disposable plastic bottles (Nunc, USA), in a humidified atmosphere of 5% CO₂

Viability results

Cell viability was not decreased in the presence of increased PTX concentrations by trypan blue assay (data not shown). MTT probe results indicated that cells decreased the mitochondrial function by 15% in the presence of PTX 150 and 170 μM and to 34% with 200 μM (data not shown). A concentration of 200 μM PTX was chosen for treatments, although this concentration decreased mitochondrial function. This concentration is 10 times lower than the one used by other groups studying collagen production

DISCUSSION

Inhibition of HSC activation and its related subsequent events such as an increase in the production of extracellular matrix components and an enhanced proliferation, are crucial goals for intervention in the hepatic fibrogenesis cascade. Our results demonstrate that, under the cell culture conditions that we employed, acetaldehyde results in an increase in collagen secretion in rat HSC. PTX, at the concentration of 200 μM, inhibited such increase in collagen secretion. This effect was mediated

Acknowledgements

This work was supported in part by the Consejo Nacional de Ciencia y Tecnologı́a (CONACYT) No. 400200-5-25175M. We thank Dr Pablo Damián Matsumura for technical advice, and Dr Rojkind by the donation of the cell line.

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PENTOXIFYLLINE DIMINISHED ACETALDEHYDE-INDUCED COLLAGEN PRODUCTION IN HEPATIC STELLATE CELLS BY DECREASING INTERLEUKIN-6 EXPRESSION

Abstract

Section snippets

INTRODUCTION

Cell culture

Viability results

DISCUSSION

Acknowledgements

J. Hepatol.

Hepatology

Gen. Pharmacol.

Biochem. Pharmacol.

J. Hepatol.

J. Immunol. Methods

Arch. Biochem. Biophys.

Anal. Biochem.

Biochem. Biophys. Acta

Cytokine

Biochem. Pharmacol.

J. Invest. Dermatol.

The cellular basis of hepatic fibrosis

New Engl. J. Med.