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Hyaluronic acid inhibits nitric oxide-induced apoptosis and dedifferentiation of articular chondrocytes in vitro

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Abstract

Objective

Nitric oxide is an important mediator in Osteoarthritis (OA), and causes apoptosis and dedifferentiation in articular chondrocytes. Protein kinase Cα is involved in modulating apoptosis and dedifferentiation of articular chondrocytes induced by nitric oxide. Hyaluronic acid is widely used in the treatment of osteoarthritis and exerts significant chondroprotective effects. The exact mechanisms of its chondroprotective action are not yet fully elucidated. The present study was performed to investigate the effects and mechanisms of hyaluronic acid in NO-induced apoptosis and dedifferentiation of chondrocytes.

Methods

The ratio of apoptotic cell and cell viability was surveyed by TUNEL, MTT assay and flow cytometry. The expression of aggrecan, type II collagen, and PKCα were determined by real-time PCR and Western blot. The expression changes of caspase-3 and bcl-2 was detected by Western blot. The mitochondrial membrane potential (ΔΨm) was evaluated by Rhodamine-123 fluorescence.

Results

HA reduces the TUNEL positive cell, nuclei fragment and the impairment of ΔΨm. NO-induced chondrocyte dedifferentiation was blocked by HA, which restores expression of aggrecan and type II collagen of chondrocytes and cell viability. HA can block inhibition of PKC-α by NO.

Conclusion

Our results show that HA blocks NO-induced apoptosis and dedifferentiation of articular chondrocytes by modulation of ΔΨm and PKCα.

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References

  1. Blanco FJ, Guitian R, Vázquez-Martul E, de Toro FJ, Galdo F. Osteoarthritis chondrocytes die by apoptosis. A possible pathway for osteoarthritis pathology. Arthritis Rheum. 1998;41:284–9.

    Article  CAS  PubMed  Google Scholar 

  2. Sandell LJ, Aigner T. Articular cartilage and changes in arthritis. An introduction: cell biology of osteoarthritis. Arthritis Res. 2001;3:107–13.

    Article  CAS  PubMed  Google Scholar 

  3. Schnabel M, Marlovits S, Eckhoff G, Fichtel I, Gotzen L, Vécsei V, et al. Dedifferentiation-associated changes in morphology and gene expression in primary human articular chondrocytes in cell culture. Osteoarthritis Cartilage. 2002;10:62–70.

    Article  CAS  PubMed  Google Scholar 

  4. Pelletier JP, Martel-Pelletier J, Abramson SB. Osteoarthritis, an inflammatory disease: potential implication for the selection of new therapeutic targets. Arthritis Rheum. 2001;44:1237–47.

    Article  CAS  PubMed  Google Scholar 

  5. Amin AR, Abramson SB. The role of nitric oxide in articular cartilage breakdown in osteoarthritis. Curr Opin Rheumatol. 1998;10:263–8.

    Article  CAS  PubMed  Google Scholar 

  6. Abramson SB, Attur M, Amin AR, Clancy R. Nitric oxide and inflammatory mediators in the perpetuation of osteoarthritis. Curr Rheumatol Rep. 2001;3:535–41.

    Article  CAS  PubMed  Google Scholar 

  7. Maneiro E, López-Armada MJ, de Andres MC, Caramés B, Martín MA, Bonilla A, et al. Effect of nitric oxide on mitochondrial respiratory activity of human articular chondrocytes. Ann Rheum Dis. 2005;64:138–95.

    Article  Google Scholar 

  8. Wu GJ, Chen TG, Chang HC, Chiu WT, Chang CC, Chen RM. Nitric oxide from both exogenous and endogenous sources activates mitochondria-dependent events and induces insults to human chondrocytes. J Cell Biochem. 2007;101:1520–31.

    Article  CAS  PubMed  Google Scholar 

  9. Cherng YG, Chang HC, Lin YL, Kuo ML, Chiu WT, Chen RM. Apoptotic insults to human chondrocytes induced by sodium nitroprusside are involved in sequential events, including cytoskeletal remodeling, phosphorylation of mitogen-activated protein kinase, kinase-1/c-Jun N-terminal kinase, and Bax-mitochondria-mediated caspase activation. J Orthop Res. 2008;26:1018–26.

    Article  CAS  PubMed  Google Scholar 

  10. Cao M, Westerhausen-Larson A, Niyibizi C, Kavalkovich K, Georgescu HI, Rizzo CF, et al. Nitric oxide inhibits the synthesis of type-II collagen without altering Col2A1 mRNA abundance: prolyl hydroxylase as a possible target. Biochem J. 1997;324:305–10.

    CAS  PubMed  Google Scholar 

  11. Abramson SB. Nitric oxide in inflammation and pain associated with osteoarthritis. Arthritis Res Ther. 2008;10:S2.

    Article  PubMed  Google Scholar 

  12. Tamura T, Nakanishi T, Kimura Y, Hattori T, Sasaki K, Norimatsu H, et al. Nitric oxide mediates interleukin-1-induced matrix degradation and basic fibroblast growth factor release in cultured rabbit articular chondrocytes: a possible mechanism of pathological neovascularization in arthritis. Endocrinology. 1996;137:3729–37.

    Article  CAS  PubMed  Google Scholar 

  13. Kim SJ, Kim HG, Oh CD, Hwang SG, Song WK, Yoo YJ, et al. p13 kinase-dependent and -independent Inhibition of protein kinase C zeta and -alpha regulates nitric oxide-induced apoptosis and dedifferentiation of articular chondrocytes. J Biol Chem. 2002;277:30375–81.

    Article  CAS  PubMed  Google Scholar 

  14. Spitaler M, Cantrell DA. Protein kinase C and beyond. Nat Immunol. 2004;5:785–90.

    Article  CAS  PubMed  Google Scholar 

  15. Yoon YM, Kim SJ, Oh CD, Ju JW, Song WK, Yoo YJ, et al. Maintenance of differentiated phenotype of articular chondrocytes by protein kinase C and extracellular signal-regulated protein kinase. J Biol Chem. 2002;277:8412–20.

    Article  CAS  PubMed  Google Scholar 

  16. Iozzo RV. Matrix proteoglycans: from molecular design to cellular function. Annu Rev Biochem. 1998;67:609–52.

    Article  CAS  PubMed  Google Scholar 

  17. Yasui T, Akatsuka M, Tobetto K, Hayaishi M, Ando T. The effect of hyaluronan on interleukin-1 alpha-induced prostaglandin E2 production in human osteoarthritic synovial cells. Agents Actions. 1992;37:155–6.

    Article  CAS  PubMed  Google Scholar 

  18. Campo GM, Avenoso A, Campo S, Ferlazzo AM, Altavilla D, Calatroni A. Efficacy of treatment with glycosaminoglycans on experimental collagen-induced arthritis in rats. Arthritis Res Ther. 2003;5:R122–31.

    Article  CAS  PubMed  Google Scholar 

  19. Sasaki A, Sasaki K, Konttinen YT, Santavirta S, Takahara M, Takei H, et al. Hyaluronate inhibits the interleukin-1 beta-induced expression of matrix metalloproteinase (MMP)-1 and MMP-3 in human synovial cells. Tohoku J Exp Med. 2004;204:99–107.

    Article  CAS  PubMed  Google Scholar 

  20. Halici M, Karaoglu S, Canoz O, Kabak S, Baktir A. Sodium hyaluronate regulating angiogenesis during Achilles tendon healing. Knee Surg Sports Traumatol Arthrosc. 2004;12:562–7.

    Article  PubMed  Google Scholar 

  21. Takahashi K, Hashimoto S, Kubo T, Hirasawa Y, Lotz M, Amiel D. Hyaluronan suppressed nitric oxide production in the meniscus and synovium of rabbit osteoarthritis model. J Orthop Res. 2001;19:500–3.

    Article  CAS  PubMed  Google Scholar 

  22. Zhou PH, Liu SQ, Peng H. The effect of hyaluronic acid on IL-1 beta-induced chondrocyte apoptosis in a rat model of osteoarthritis. J Orthop Res. 2008;26:1643–8.

    Article  CAS  PubMed  Google Scholar 

  23. Chen Q, Liu SQ, Du YM, Peng H, Sun LP. Carboxymethyl-chitosan protects rabbit chondrocytes from interleukin-1 beta-induced apoptosis. Eur J Pharmacol. 2006;54:1–8.

    Google Scholar 

  24. Johnson EO, Charchandi A, Babis GC, Soucacos PN. Apoptosis in osteoarthritis: morphology, mechanisms, and potential means for therapeutic intervention. J Surg Orthop Adv. 2008;17:147–52.

    PubMed  Google Scholar 

  25. Kim HA, Blanco FJ. Cell death and apoptosis in osteoarthritic cartilage. Curr Drug Targets. 2007;8:333–45.

    Article  CAS  PubMed  Google Scholar 

  26. Hashimoto S, Ochs RL, Komiya S, Lotz M. Linkage of chondrocyte apoptosis and cartilage degradation in human osteoarthritis. Arthritis Rheum. 1998;41:1632–8.

    Article  CAS  PubMed  Google Scholar 

  27. Farrell AJ, Blake DR, Palmer RM, Moncada S. Increased concentrations of nitrite in synovial fluid and serum samples suggest increased nitric oxide synthesis in rheumatic diseases. Ann Rheum Dis. 1992;51:1219–22.

    Article  CAS  PubMed  Google Scholar 

  28. Taskiran D, Stefanovic-Racic M, Georgescu HI, Evans C. Nitric oxide mediates suppression of cartilage proteoglycan synthesis by interleukin-1. Biochem Biophys Res Commun. 1994;200:142–8.

    Article  CAS  PubMed  Google Scholar 

  29. Kim HA, Lee YJ, Seong SC, Choe KW, Song YW. Apoptotic chondrocyte death in human osteoarthritis. J Rheumatol. 2000;27:455–62.

    CAS  PubMed  Google Scholar 

  30. Renoux M, Hiliquin P, Galoppin L, Florentin I, Menkes CJ. Release of mast cell mediators and nitrites into knee joint fluid in osteoarthritis—comparison with articular chondrocalcinosis and rheumatoid arthritis. Osteoarthritis Cartilage. 1996;4:175–9.

    Article  CAS  PubMed  Google Scholar 

  31. Kösel S, Hofhaus G, Maassen A, Vieregge P, Graeber MB. Role of mitochondria in Parkinson disease. Biol Chem. 1999;130:865–70.

    Article  Google Scholar 

  32. Leonard JV, Schapira AH. Mitochondrial respiratory chain disorders I: mitochondrial DNA defects. Lancet. 2000;355:299–304.

    Article  CAS  PubMed  Google Scholar 

  33. Green DR, Reed JC. Mitochondria and apoptosis. Science. 1998;281:1309–12.

    Article  CAS  PubMed  Google Scholar 

  34. Petit PX, Zamzami N, Vayssière JL, Mignotte B, Kroemer G, Castedo M. Implication of mitochondria in apoptosis. Mol Cell Biochem. 1997;174:185–8.

    Article  CAS  PubMed  Google Scholar 

  35. López-Armada MJ, Caramés B, Martín MA, Cillero-Pastor B, Lires-Dean M, Fuentes-Boquete I, et al. Mitochondrial activity is modulated by TNFα and IL-1β in normal human chondrocyte cells. Osteoarthritis Cartilage. 2006;14:1011–22.

    Article  PubMed  Google Scholar 

  36. Johnson K, Jung A, Murphy A, Andreyev A, Dykens J, Terkeltaub R. Mitochondrial oxidative phosphorylation is a downstream regulator of nitric oxide effects on chondrocyte matrix synthesis and mineralization. Arthritis Rheum. 2000;43:1560–70.

    Article  CAS  PubMed  Google Scholar 

  37. Tomita M, Sato EF, Nishikawa M, Yamano Y, Inoue M. Nitric oxide regulates mitochondrial respiration and functions of articular chondrocytes. Arthritis Rheum. 2001;44:96–104.

    Article  CAS  PubMed  Google Scholar 

  38. Grishko V, Xu M, Ho R, Mates A, Watson S, Kim JT, et al. Effects of hyaluronic acid on mitochondrial function and mitochondria-driven apoptosis following oxidative stress in human chondrocytes. J Biol Chem. 2009;284:9132–9.

    Article  CAS  PubMed  Google Scholar 

  39. Kim SJ, Hwang SG, Shin DY, Kang SS, Chun JS. P38 kinase regulates nitric oxide-induced apoptosis of articular chondrocytes by accumulating p53 via NFkappa B-dependent transcription and stabilization by serine 15 phosphorylation. J Biol Chem. 2002;277:33501–8.

    Article  CAS  PubMed  Google Scholar 

  40. Kim SJ, Ju JW, Oh CD, Yoon YM, Song WK, Kim JH, et al. ERK-1/2 and p38 kinase oppositely regulate nitric oxide-induced apoptosis of chondrocytes in association with p53, caspase-3, and differentiation status. J Biol Chem. 2002;277:1332–9.

    Article  CAS  PubMed  Google Scholar 

  41. Qiu B, Liu SQ, Peng H. Influence of sodium hyaluronate on iNOS expression in synovium and NO content in synovial fluid of rabbits with traumatic osteoarthritis. Chin J Traumatol. 2008;11:293–6.

    Article  CAS  PubMed  Google Scholar 

  42. Kalaci A, Yilmaz HR, Aslan B, Söğüt S, Yanat AN, Uz E. Effects of hyaluronan on nitric oxide levels and superoxide dismutase activities in synovial fluid in knee osteoarthritis. Clin Rheumatol. 2007;26:1306–11.

    Article  PubMed  Google Scholar 

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Acknowledgments

This work was supported by fund from the Health Department (No.JX4B15) and Science & Technology Department (No.2009CBD034) of Hubei Province of China.

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Authors and Affiliations

  1. Department of Orthopedics, Renmin Hospital of Wuhan University, 430060, Wuhan, Hubei, People’s Republic of China

    Hao Peng, Jian-lin Zhou, Shi-qing Liu, Qiong-jie Hu, Jiang-hua Ming & Bo Qiu

  2. Department of Respiratory Medicine, Tongji Hospital, Tongji Medical College, Key Laboratory of Respiratory Disease of Ministry of Public Health, Huazhong University of Science and Technology, 430030, Wuhan, Hubei, People’s Republic of China

    Qiong-jie Hu

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  1. Hao Peng
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Correspondence to Jian-lin Zhou.

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Responsible Editor: J. Di Battista.

H. Peng, J. Zhou, S. Liu, Q. Hu, J. Ming and B. Qiu contributed equally to this work.

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Peng, H., Zhou, Jl., Liu, Sq. et al. Hyaluronic acid inhibits nitric oxide-induced apoptosis and dedifferentiation of articular chondrocytes in vitro. Inflamm. Res. 59, 519–530 (2010). https://doi.org/10.1007/s00011-010-0156-x

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  • DOI: https://doi.org/10.1007/s00011-010-0156-x

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