Biomedical Science Letters (대한의생명과학회지)

The Korean Society for Biomedical Laboratory Sciences (대한의생명과학회)
권호 표지

A77 1726 Inhibit NO-induced Apoptosis via PI-3K/AKT Signaling Pathway in Rabbit Articular Chondrocyte

  • Choi, In-Kyou (Department of Biological Sciences, College of Natural Sciences, Kongju National University) ;
  • Kim, Song-Ja (Department of Biological Sciences, College of Natural Sciences, Kongju National University)
  • ;
  • 김송자 (국립공주대학교 자연과학대학 생명과학과)
  • Published : 2009.03.31
Download PDF
⟨ Previous Next ⟩

Abstract

Leflunomide is an immunomodulatory agent used for the treatment of rheumatoid arthritis (RA). Leflunomide known as a regulator of iNOS synthesis which largely decreases NO production in diverse cell type. However, the effect of leflunomide on chondrocyte is still poorly understood. In our previous studies, we have shown that direct production of Nitric oxide (NO) by treating chondrocytes with NO donor, sodium nitroprusside (SNP), causes apoptosis via p38 mitogen-activated protein kinase in association with elevation of p53 protein level, caspase-3 activation. In this study, we characterized the molecular mechanism by which A77 1726 inhibit apoptosis. We found that A77 1726 inhibit NO-induced apoptosis as determined by MTT (Thiazolyl Blue Tetrazolium Bromide) assay and DNA fragmentation. The inhibition of apoptosis by A77 1726 was accompanied by increased PI-3 kinase and AKT activities. So, inhibition of phosphatidylinositol (PI)-3kinase with LY294002 rescued apoptosis. Triciribine, the specific inhibitor of AKT, also abolished anti-apoptotic effect. Our results indicate that A77 1726, the active metabolite of leflunomide, mediates NO-induced apoptosis in chondrocytes by modulating up-regulation of PI-3 kinase and AKT.

Keywords

References

  1. Amin AR, Abramson SB. The role of nitric oxide in articular cartilage breakdown in osteoarthritis. Curr Opin Rheumatol. 1998. 10: 263-268. https://doi.org/10.1097/00002281-199805000-00018
  2. Blanco FJ, Guitian R. Osteoarthritis chondrocytes die by apoptosis. A possible pathway for osteoarthritis pathology. Arthritis Rheum. 1998. 41: 284-289. https://doi.org/10.1002/1529-0131(199802)41:2<284::AID-ART12>3.0.CO;2-T
  3. Breedveld FC, Dayer JM. Leflunomide: mode of action in the treatment of rheumatoid arthritis. Ann Rheum Dis. 2000. 59: 841-849. https://doi.org/10.1136/ard.59.11.841
  4. Cash DE, Bock CB. Retinoic acid receptor alpha function in vertebrate limb skeletogenesis: a modulator of chondrogenesis. J Cell Biol. 1997. 136: 445-457. https://doi.org/10.1083/jcb.136.2.445
  5. DeLise AM, Fischer L. Cellular interactions and signaling in cartilage development. Osteoarthritis Cartilage. 2000. 8: 309 -334. https://doi.org/10.1053/joca.1999.0306
  6. Emery P, Breedveld FC. A comparison of the efficacy and safety of leflunomide and methotrexate for the treatment of rheumatoid arthritis. Rheumatology (Oxford) 2000. 39: 655-665. https://doi.org/10.1093/rheumatology/39.6.655
  7. Fox RI, Herrmann ML. How does leflunomide modulate the immune response in rheumatoid arthritis- Bio Drugs 1999. 12: 301-315.
  8. Goldring MB, Birkhead JR. Interleukin-1 beta-modulated gene expression in immortalized human chondrocytes. J Clin Invest. 1994. 94: 2307-2316. https://doi.org/10.1172/JCI117595
  9. Hashimoto SR, Ochs L. Linkage of chondrocyte apoptosis and cartilage degradation in human osteoarthritis. Arthritis Rheum. 1998. 41: 1632-1638. https://doi.org/10.1002/1529-0131(199809)41:9<1632::AID-ART14>3.0.CO;2-A
  10. Herrmann ML, Schleyerbach R. Leflunomide: an immunomodulatory drug for the treatment of rheumatoid arthritis and other autoimmune diseases. Immunopharmacology 2000. 47: 273-289. https://doi.org/10.1016/S0162-3109(00)00191-0
  11. Kim JS, Park ZY. p38 kinase mediates nitric oxide-induced apoptosis of chondrocytes through the inhibition of protein kinase C zeta by blocking autophosphorylation. Cell Death Differ. 2005. 12: 201-212. https://doi.org/10.1038/sj.cdd.4401511
  12. Kim SJ, Hwang SG. 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: 33501-33508. https://doi.org/10.1074/jbc.M202862200
  13. Kim SJ, Ju JW. 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: 1332-1339. https://doi.org/10.1074/jbc.M107231200
  14. Lefebvre V, Peeters-Joris C. Production of collagens, collagenase and collagenase inhibitor during the dedifferentiation of articular chondrocytes by serial subcultures. Biochim Biophys Acta. 1990. 1051: 266-275. https://doi.org/10.1016/0167-4889(90)90132-W
  15. Miljkovic D, Samardzic T. Leflunomide inhibits activation of inducible nitric oxide synthase in rat astrocytes. 2001. Brain Res. 889: 331-338. https://doi.org/10.1016/S0006-8993(00)03181-4
  16. Pelletier JP, Fernandes JC. Chondrocyte death in experimental osteoarthritis is mediated by MEK 1/2 and p38 pathways: role of cyclooxygenase-2 and inducible nitric oxide synthase. J Rheumatol. 2001. 28: 2509-2519.
  17. Sandell LJ, Adle PR. Developmental patterns of cartilage. Front Biosci. 1999. 4: D731-742. https://doi.org/10.2741/Sandell
  18. Smolen JS, Kalden JR. Efficacy and safety of leflunomide compared with placebo and sulphasalazine in active rheumatoid arthritis: a double-blind, randomised, multicentre trial. European Leflunomide Study Group. Lancet 1999. 353: 259-266. https://doi.org/10.1016/S0140-6736(98)09403-3
  19. Strand VS, Cohen. Treatment of active rheumatoid arthritis with leflunomide compared with placebo and methotrexate. Leflunomide Rheumatoid Arthritis Investigators Group. Arch Intern Med. 1999. 159: 2542-2550. https://doi.org/10.1001/archinte.159.21.2542
  20. Weston AD, Rosen V. Regulation of skeletal progenitor differentiation by the BMP and retinoid signaling pathways. J Cell Biol. 2000. 148: 679-690. https://doi.org/10.1083/jcb.148.4.679
  21. Yatsugi N, Tsukazaki T. Apoptosis of articular chondrocytes in rheumatoid arthritis and osteoarthritis: correlation of apoptosis with degree of cartilage destruction and expression of apoptosis-related proteins of p53 and c-myc. J Orthop Sci. 2000. 5: 150-156. https://doi.org/10.1007/s007760050142
  22. Yoon YM, Kim SJ. Maintenance of differentiated phenotype of articular chondrocytes by protein kinase C and extracellular signal-regulated protein kinase. J Biol Chem. 2002. 277: 8412 -8420. https://doi.org/10.1074/jbc.M110608200