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Cyclic compression-induced p38 activation and subsequent MMP13 expression requires Rho/ROCK activity in bovine cartilage explants

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Abstract

Objective

Excessive mechanical stress on the cartilage causes the degradation of the matrix, leading to the osteoarthritis (OA). Matrix metalloproteinases 13 (MMP13) is a major catalytic enzyme in OA and p38 plays an important role in its induction. However, precise pathway inducing p38 activation has not been elucidated. We hypothesized here that the small GTPase Rho and its effector ROCK might function in upper part of the mechanical stress-induced matrix degeneration pathway.

Methods

Bovine metacarpal phalangeal articular cartilage explants were loaded with 1 MPa dynamic compression for 6 h with or without a ROCK specific inhibitor Y27632 or/and a p38 specific inhibitor SB202190. Then p38 phosphorylation and MMP13 expression were assessed by western blot or/and quantitative RT-PCR. Rho-activity was measured by pull-down assay using glutathione S-transferase fusion protein of Rho binding domain.

Results

Cyclic compression caused Rho activation, p38 phosphorylation and MMP13 expression. Both Y27632 and SB202190 were found to block the mechanical stress-enhanced p38 phosphorylation and subsequent MMP13 expression.

Conclusions

The present results show that p38 phosphorylation and MMP13 expression are regulated by Rho/ROCK activation, and support the potential novel pathway that Rho/ROCK is in the upper part of the mechanical stress-induced matrix degeneration cascade in cartilage comprised of p38 and MMP13.

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Acknowledgments

We gratefully acknowledge Ms. Naomi Backes Kamimura, Department of Biology-Oriented Science and Technology, Kinki University, for English editing. We also thank Ms. Kanae Shigi and Ms. Naoko Ohoshi for excellent technical assistance.

Author information

Authors and Affiliations

  1. Department of Orthopaedic Surgery, Kinki University Faculty of Medicine, Osaka, Japan

    Koichi Nakagawa, Chiaki Hamanishi, Masao Akagi & Kanji Fukuda

  2. Division of Cell Biology for Regenerative Medicine, Institute of Advanced Clinical Medicine, Kinki University Faculty of Medicine, 377-2 Ohno-higashi, Osaka-sayama, Osaka, 5898511, Japan

    Takeshi Teramura, Toshiyuki Takehara, Yuta Onodera & Kanji Fukuda

  3. Department of Rehabilitation Medicine, Kinki University Faculty of Medicine, Osaka, Japan

    Kanji Fukuda

Authors
  1. Koichi Nakagawa
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  2. Takeshi Teramura
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Correspondence to Takeshi Teramura.

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Responsible Editor: John Di Battista.

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11_2012_500_MOESM1_ESM.tif

Supplement 1. Other ROCK inhibitor fasudil also blocked the cyclic compression induced p38 phosphorylation and MMP13 expression. A. Western blot analysis for p38 and MMP13. B. qRT-PCR analysis for MMP13. Both SB202190 or fasudil (ROCK inhibitor) supplementation during compression treatment blocked mechanical stress induced MMP13 expression. Asterisk means significant differences at p < 0.05. All experimental groups included the vehicle of SB202190 (DMSO) at same concentration. Bars show the mean score of three independent experiments and bars depict S.D. (TIFF 3435 kb)

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Nakagawa, K., Teramura, T., Takehara, T. et al. Cyclic compression-induced p38 activation and subsequent MMP13 expression requires Rho/ROCK activity in bovine cartilage explants. Inflamm. Res. 61, 1093–1100 (2012). https://doi.org/10.1007/s00011-012-0500-4

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  • DOI: https://doi.org/10.1007/s00011-012-0500-4

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