Skip to main content

Advertisement

Log in

TNFα-mediated apoptosis in human osteoarthritic chondrocytes sensitized by PI3K-NF-κB inhibitor, not mTOR inhibitor

  • Original Article
  • Published:
Rheumatology International Aims and scope Submit manuscript

Abstract

To investigate apoptosis of osteoarthritic (OA) chondrocytes stimulated with different inhibitors targeting tumor necrosis factor-alpha (TNFα) pathway, we isolated first passage chondrocytes from OA patients and then treated them with the inhibitors in combination with TNFα, and then collected the stimulated chondrocytes for Western blotting. Chondrocytes from OA patients expressed cleaved caspase-3 and PARP, suggesting apoptotic background. We here, validated that 10 ng/ml of TNFα couldn’t induce more chondrocytes apoptosis. PI3K inhibitor LY294002 or NF-κB inhibitor CAPE, but not mTOR inhibitor rapamycin and MEK1/2 inhibitor U0126 in combination with TNFα could facilitate apoptosis. CAPE-induced more apoptosis could be explained by c-FLIP downregulation more than cIAP1 upregulation. And, we showed the first time that PI3K-NF-κB pathway, but not mTOR pathway could prevent chondrocytes apoptosis induced by a pro-apoptotic factor TNFα and call for attention while trying to inhibit NF-κB as a therapeutic target.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Felson DT (2008) Comparing the prevalence of rheumatic disease in China with the rest of the world. Arthr Res Ther 10(1):1–2

    Article  Google Scholar 

  2. Samuels J, Krasnokutsky S, Abramson SB (2008) Osteoarthritis: a tale of three tissues. Bull NYU Hosp Joint Dis 66(3):244–250

    Google Scholar 

  3. Carames B, Lopez-Armada MJ, Cillero-Pastor B, Lires-Dean M, Vaamonde C, Galdo F et al (2008) Differential effects of tumor necrosis factor-alpha and interleukin-1beta on cell death in human articular chondrocytes. Osteoarthr Cartil 16(6):715–722

    Article  PubMed  CAS  Google Scholar 

  4. John T, Muller RD, Oberholzer A, Zreiqat H, Kohl B, Ertel W et al (2007) Interleukin-10 modulates pro-apoptotic effects of TNF-alpha in human articular chondrocytes in vitro. Cytokine 40(3):226–234

    Article  PubMed  CAS  Google Scholar 

  5. Lee SW, Lee HJ, Chung WT, Choi SM, Rhyu SH, Kim DK et al (2004) TRAIL induces apoptosis of chondrocytes and influences the pathogenesis of experimentally induced rat osteoarthritis. Arthritis Rheum 50(2):534–542

    Article  PubMed  CAS  Google Scholar 

  6. Seigio S (2008) Increased levels of apo-A1 and apo B are associated in knee osteoarthritis: lack of association with VEGF-460 T/C and +405 C/G polymorphisms. Rheumatol Int 29:63–68

    Article  Google Scholar 

  7. Mounach A (2008) Risk factors for knee osteoarthritis in Morocco. A case control study. Clin Rheumatol 27:323–326

    Article  PubMed  Google Scholar 

  8. Lalichman K (2007) Age, body composition, and reproductive indices as predictors of radiographic hand osteoarthritis in Chuvashian women. Scand J Rheumatol 36:53–57

    Article  Google Scholar 

  9. Dahaghin S (2007) Do metabolic factors add to the effect of overweight on hand osteoarthritis? The Rotterdam study. Ann Rheum Dis 66:916–920

    Article  PubMed  CAS  Google Scholar 

  10. Nieto-Vazquez I, Fernandez-Veledo S, Kramer DK (2008) Insulin resistance associated to obesity: the link TNF-alpha. Archives Physiol Biochem 114(3):183–194

    Google Scholar 

  11. Hussein MR, Fathi NA, El-Din AM, Hassan HI, Abdullah F, Al-Hakeem E et al (2008) Alterations of the CD4(+), CD8(+) T cell subsets, interleukins-1beta, IL-10, IL-17, tumor necrosis factor-alpha and soluble intercellular adhesion molecule-1 in rheumatoid arthritis and osteoarthritis: preliminary observations. Pathol Oncol Res 14(3):321–328

    Article  PubMed  CAS  Google Scholar 

  12. Hulejova H, Baresova V, Klezl Z, Polanska M, Adam M, Senolt L (2007) Increased level of cytokines and matrix metalloproteinases in osteoarthritic subchondral bone. Cytokine 38(3):151–156

    Article  PubMed  CAS  Google Scholar 

  13. Penninx BW, Abbas H, Ambrosius W, Nicklas BJ, Davis C, Messier SP et al (2004) Inflammatory markers and physical function among older adults with knee osteoarthritis. J Rheumatol 31(10):2027–2031

    PubMed  Google Scholar 

  14. Urech DM (2010) Anti-inflammatory and cartilage-protecting efffects of an intra-articularly injected anti-TNFa single-chain Fv antibody (ESBA105) designed for local therapeutic use. Ann Rheum Dis 69:443–449

    Article  PubMed  CAS  Google Scholar 

  15. Clohisy JC, Roy BC, Biondo C, Frazier E, Willis D, Teitelbaum SL et al (2003) Direct inhibition of NF-kappaB blocks bone erosion associated with inflammatory arthritis. J Immunol 171(10):5547–5553

    PubMed  CAS  Google Scholar 

  16. Lianxu C, Hongti J, Changlong Y (2006) NF-kappaB p65-specific siRNA inhibits expression of genes of COX-2, NOS-2 and MMP-9 in rat IL-1beta-induced and TNF-alpha-induced chondrocytes. Osteoarthr Cartil 14(4):367–376

    Article  PubMed  CAS  Google Scholar 

  17. Chen LX, Lin L, Wang HJ, Wei XL, Fu X, Zhang JY et al (2008) Suppression of early experimental osteoarthritis by in vivo delivery of the adenoviral vector-mediated NF-kappaB p65-specific siRNA. Osteoarthr Cartil 16(2):174–184

    Article  PubMed  CAS  Google Scholar 

  18. Yao Z, Xing L, Boyce BF (2009) NF-kappaB p100 limits TNF-induced bone resorption in mice by a TRAF3-dependent mechanism. J Clin Invest 119(10):3024–3034

    Article  PubMed  CAS  Google Scholar 

  19. Yoon HS, Kim HA, Song YW (2006) Inhibition of NF-kappaB renders human juvenile costal chondrocyte cell lines sensitive to TNF-alpha-mediated cell death. Rheumatol Int 26(3):201–208

    Article  PubMed  CAS  Google Scholar 

  20. Wang L, Du F, Wang X (2008) TNF-alpha induces two distinct caspase-8 activation pathways. Cell 133(4):693–703

    Article  PubMed  CAS  Google Scholar 

  21. Cejka D, Hayer S, Niederreiter B, Sieghart W, Fuereder T, Zwerina J et al. (2010) mTOR signaling is crucial for joint destruction in TNF-alpha-driven arthritis. Arthritis Rheum. doi:10.1002/art.27504

  22. Kwiatkowski DJ, Zhang H, Bandura JL, Heiberger KM, Glogauer M, el-Hashemite N et al (2002) A mouse model of TSC1 reveals sex-dependent lethality from liver hemangiomas, and up-regulation of p70S6 kinase activity in Tsc1 null cells. Hum Mol Genet 11(5):525–534

    Article  PubMed  CAS  Google Scholar 

  23. Kim HA, Lee YJ, Seong SC, Choe KW, Song YW (2000) Apoptotic chondrocyte death in human osteoarthritis. J Rheumatol 27(2):455–462

    PubMed  CAS  Google Scholar 

  24. Blanco FJ, Guitian R, Vazquez-Martul E, de Toro FJ. Galdo F (1998) Osteoarthritis chondrocytes die by apoptosis. A possible pathway for osteoarthritis pathology. Arthritis Rheum 41(2):284–289

    Google Scholar 

  25. James AF (2007) IAP antagonists target cIAP1 to induce TNFa-dependent apoptosis. Cell 131:682–693

    Article  Google Scholar 

  26. Seguin CA, Bernier SM (2003) TNFalpha suppresses link protein and type II collagen expression in chondrocytes: role of MEK1/2 and NF-kappaB signaling pathways. J Cell Physiol 197(3):356–369

    Article  PubMed  Google Scholar 

  27. Wullschleger S, Loewith R, Hall MN (2006) TOR signaling in growth and metabolism. Cell 124(3):471–484

    Article  PubMed  CAS  Google Scholar 

  28. Sugatani T, Hruska KA (2005) Akt1/Akt2 and mammalian target of rapamycin/Bim play critical roles in osteoclast differentiation and survival, respectively, whereas Akt is dispensable for cell survival in isolated osteoclast precursors. J Biol Chem 280(5):3583–3589

    Article  PubMed  CAS  Google Scholar 

  29. Phornphutkul C, Lee M, Voigt C, Wu KY, Ehrlich MG, Gruppuso PA et al (2009) The effect of rapamycin on bone growth in rabbits. J Orthop Res 27(9):1157–1161

    Article  PubMed  CAS  Google Scholar 

  30. Janus A, Linke A, Cebula B, Robak T, Smolewski P (2009) Rapamycin, the mTOR kinase inhibitor, sensitizes acute myeloid leukemia cells, HL-60 cells, to the cytotoxic effect of arabinozide cytarabine. Anticancer Drugs 20(8):693–701

    Article  PubMed  CAS  Google Scholar 

  31. Varfolomeev EE, Ashkenazi A (2004) Tumor necrosis factor: an apoptosis JuNKie? Cell 116(4):491–497

    Article  PubMed  CAS  Google Scholar 

  32. Relic B, Bentires-Alj M, Ribbens C, Franchimont N, Guerne PA, Benoit V et al (2002) TNF-alpha protects human primary articular chondrocytes from nitric oxide-induced apoptosis via nuclear factor-kappaB. Lab Invest 82(12):1661–1672

    PubMed  CAS  Google Scholar 

  33. Xu J, Wu HF, Ang ES, Yip K, Woloszyn M, Zheng MH et al (2009) NF-kappaB modulators in osteolytic bone diseases. Cytokine Growth Factor Rev 20(1):7–17

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

We thank Wu Gang, Xue Hong and Ma Jianhui for some experiment skills support.

Conflict of interest

The authors declare that they have no conflict of interest.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Guixing Qiu.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Li, D., Wu, Z., Duan, Y. et al. TNFα-mediated apoptosis in human osteoarthritic chondrocytes sensitized by PI3K-NF-κB inhibitor, not mTOR inhibitor. Rheumatol Int 32, 2017–2022 (2012). https://doi.org/10.1007/s00296-011-1929-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00296-011-1929-4

Keywords

Navigation