PKCa Agonists Enhance the Protective Effect of Hyaluronic Acid on Nitric Oxide-Induced Apoptosis of Articular Chondrocytes in Vitro

Document Type: Original Article

Authors

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

2 Department of Radiology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei Province, People’s Republic of China

Abstract

 




Objective(s):
Protein kinase C (PKCα) is involved in modulating articular chondrocytes apoptosis induced by nitric oxide (NO). Hyaluronic acid (HA) inhibits nitric oxide-induced apoptosis of articular chondrocytes by protecting PKCα, but the mechanism remains unclear. The present study was performed to investigate the effects and mechanisms of PKCα regulate protective effect of hyaluronic acid.
 
Materials and Methods
The ratio of apoptotic cell and cell viability was surveyed by PCNA and MTT assay. The expression of caspase-3 was determined by real-time PCR and western blot.
Results:
It was showed that HA was able to reduce the nuclei fragment and PCNA expression, and NO-induced articular apoptosis blocked by HA, pretreated chondrocytes with PMA, HA significantly inhibits the activation of caspase-3 induced by NO, but pretrement with CHR, HA significantly incresed the expression of caspase-3.
Conclusion:
The results may be showed that PKCa regulate the expresion of caspase-3, which contribute to the apoptosis of chondrocytes induced by NO. PKC α agonists enhance the protective effect of hyaluronic acid on nitric oxide-induced articular chondrocytes apoptosis.

Keywords


 

1. Farrell AJ, Blake DR, Palmer RMJ, Moncada S. Increased concentrations of nitrite in synovial fluid and serum samples suggest increased nitric oxide synthesis in rheumatic diseases. Ann Rheum Dis 1992; 51:1219–1222.

2. Taskiran D, Stefanovic-Racic M, Georgescu HI, Evans CH. Nitric oxide mediates suppression of cartilage proteoglycan synthesis by interleukin-1. Biochem Biophys Res Commun 1994; 200:142–148.

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

4. Renoux M, Hilliquin 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; 4:175–179.

5. Kim SJ, Ju JW, Oh CD, Yoon YM, Song WK, Kim JH, Yoo YJ, Bang OS, Kang SS, Chun JS

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; 277:1332-1339.

6. Kim SJ, Kim HG, Oh CD, Hwang SG, Song WK, Yoo YJ,

et al. P38 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; 277:30375–30381.

7. 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; 277:33501–33508.

8. Iozzo RV. Matrix proteoglycans: from molecular design to cellular function. Ann Rev Biochem 1998; 67:609-652.

9. 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

PKCa Regulate Chondrocytes Apoptosis Induced by NO Zhou et al

Iran J Basic Med Sci, Vol. 16, No. 12, Dec 2013

 

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synovial cells. Agents Actions 1992; 37:155–156.

10. 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:R122-131.

11. Sasaki A, Sasaki K, Konttinen YT, Santavirta S, Takahara M, Takei H,

et al. Hyaluronate inhibits the interleukin-1beta-induced expression of matrix metalloproteinase (MMP)-1 and MMP-3 in human synovial cells. Tohoku J Exp Med 2004; 204:99-107.

12. Halici M, Karaoglu S, Canoz O, Kabak S, Baktir A. Sodium hyaluronate regulating angiogenesis during Achilles tendon healing.Knee Surg Sports Traumatol Arthrosc 2004; 12:562-567.

13. 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; 19:500-503.

14. Peng H, Zhou JL, Liu SQ, Hu QJ, Ming JH, Qiu B. Hyaluronic acid inhibits nitric oxide-induced apoptosis and dedifferentiation of articular chondrocytes

in vitro. Inflamm Res 2010; 59:519–530.

15. Matassa AA, Kalkofen RL, Carpenter L, Biden TJ, Reyland ME. Inhibition of PKCalpha induces a PKCdelta-dependent apoptotic program in salivary epithelial cells. Cell Death Differ 2003; 10:269–277.

16. Loeser RF, Carlson CS, Del Carlo M, Cole A. Detection of nitrotyrosine in aging and osteoarthritic cartilage:Correlation of oxidative damage with the

presence of interleukin-1beta and with chondrocyte resistance to insulin-like growth factor 1. Arthritis Rheum 2002; 46:2349–2357.

17. Boileau C, Martel-Pelletier J, Moldovan F, Jouzeau JY, Netter P, Manning PT

, et al. The in situ up-regulation of chondrocyte interleukin-1-converting enzyme and interleukin-18 levels in experimental osteoarthritis is mediated by nitric oxide. Arthritis Rheum 2002; 46:2637–2647.

18. 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 hondrocytes. J Cell Biochem 2007; 101:1520–1531.

19. Maneiro E, Lopez-Armada MJ, de Andres MC, Carames B, Martin MA, Bonilla A,

et al. Effect of nitric oxide on mitochondrial respiratory activity of human articular chondrocytes. Ann Rheum Dis 2005; 64:388–395.

20. Steven B. Abramson Osteoarthritis and nitric oxide. Osteoarthritis Cartilage 2008; 16: S15–S20.

21. 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; 27:1332-1339.

22. 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; 11:293-296.