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Methotrexate modulates the expression of MMP-1 and type 1 collagen in dermal fibroblast

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Abstract

Methotrexate (MTX), an anti-metabolite and anti-inflammatory drug, has been used to effectively manage and prevent keloids, but its mechanism(s) of action has not been elucidated. Our study sought to evaluate the effect of MTX on the production of key extra cellular matrix components, collagen, and matrix metalloproteinase-1 (MMP-1), produced by fibroblasts and involved in development of fibrosis. The proliferation and viability of cultured human dermal fibroblasts in response to different concentrations of MTX were determined using cell counting and MTT assay, respectively. Western blot analysis was used to determine the levels of both intracellular and secreted type 1 collagen and MMP-1. The results showed no significant changes in the proliferation of fibroblasts treated with 50 ng/ml of MTX as compared to that of control. Under the same experimental conditions, the level of secreted and intracellular type I collagen was markedly reduced and, conversely, the level of MMP-1 increased in treated neonatal, adult, and hypertrophic scar fibroblasts as compared with those of controls. The possible involvement of MTX-induced extracellular signal-regulated kinase 1/2 (ERK1/2) pathway in MMP-1 production was also studied and the result showed an increase in phosphorylated ERK 1/2 in response to MTX treatment. In summary, the findings of this study revealed that MTX significantly reduced collagen production in different strains of fibroblasts derived from neonatal, adult, and hypertrophic scar tissues, while under the same experimental conditions, it increased the expression of MMP-1. As such, our findings validate and identify a potential mechanism through which MTX functions as an anti-fibrogenic factor in treating fibroproliferative disorders.

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Acknowledgments

This research was funded by the Canadian Institute of Health Research (AG) as well as a generous support from the WorkSafe BC, International Brotherhood Electrical Workers and Spectra Energy.

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Author notes

  1. Aziz Ghahary

    Present address: The Blusson Spinal Cord Centre, 818 West 10th Ave., Vancouver, BC, V5Z1M9, Canada

Authors and Affiliations

  1. BC Professional Firefighters’ Burn and Wound Healing Research Laboratory, Department of Surgery/Plastic Surgery, University of British Columbia, Vancouver, Canada

    Layla Nabai, Ruhangiz T. Kilani, Farzian Aminuddin, Yunyuan Li & Aziz Ghahary

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  1. Layla Nabai
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  2. Ruhangiz T. Kilani
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Correspondence to Aziz Ghahary.

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Nabai, L., Kilani, R.T., Aminuddin, F. et al. Methotrexate modulates the expression of MMP-1 and type 1 collagen in dermal fibroblast. Mol Cell Biochem 409, 213–224 (2015). https://doi.org/10.1007/s11010-015-2526-8

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  • DOI: https://doi.org/10.1007/s11010-015-2526-8

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