BENEFICIAL OUTCOMES AFTER COMBINED THERAPY WITH AZATHIOPRINE AND PIOGLITAZONE IN PRECLINICAL ARTHRITIC MODELS
Objectives: The present study was designed to investigate the antioxidant and anti-arthritic activities of azathioprine-pioglitazone combination therapy to check whether it has an add on benefit over monotherapy with azathioprine or pioglitazone on disease activity in rodent models.
Methods: The antioxidant activity of test drugs and their combination was screened by H2O2 and nitric oxide scavenging assays. They were further evaluated for anti-arthritic activity using in vitro models such as protein denaturation and membrane stabilization and in vivo methods such as formaldehyde and complete Freund’s adjuvant (CFA)-induced arthritis.
Results: The combination of test drugs showed better inhibition of free radicals in both H2O2 and nitric oxide scavenging assay than individual counterparts revealing its potential antioxidant activity. They also showed significant inhibition of protein denaturation and proliferation of the red blood cell in in vitro models. The test drugs showed significant inhibition of the paw volume in both the formaldehyde and CFA-induced arthritis along with reverting the altered biochemical parameters. These findings were corroborated by radiological and histopathological studies.
Conclusions: Combination of azathioprine and pioglitazone exhibited better antioxidant and anti-arthritic effect than the individual drugs showing synergistic interaction between them.
2. Fotina AA, Fisinin I, Surai PF. Recent developments in usage of natural antioxidants to improve chicken meat production and quality. Bulg J Agric Sci 2013;19:889-96.
3. Hadjigogos K. The role of free radicals in the pathogenesis of rheumatoid arthritis. Panminerva Med 2003;45:7-13.
4. Klareskog L, Catrina AI, Paget S. Rheumatoid arthritis. Lancet 2009;373:659-72.
5. Symmons D, Turner G, Webb R, Asten P, Barrett E, Lunt M, et al. The prevalence of rheumatoid arthritis in the United Kingdom: New estimates for a new century. Rheumatology (Oxford) 2002;41:793-800.
6. Englund M, Jöud A, Geborek P, Felson DT, Jacobsson LT, Petersson IF, et al. Prevalence and incidence of rheumatoid arthritis in Southern Sweden 2008 and their relation to prescribed biologics. Rheumatology (Oxford) 2010;49:1563-9.
7. Siebert S, Tsoukas A, Robertson J, McInnes I. Cytokines as therapeutic targets in rheumatoid arthritis and other inflammatory diseases. Pharmacol Rev 2015;67:280-309.
8. Firestein GS. Evolving concepts of rheumatoid arthritis. Nature 2003;423:356-61.
9. Lee DM, Weinblatt ME. Rheumatoid arthritis. Lancet 2001;358:903-11.
10. Gibofsky A. Combination therapy for rheumatoid arthritis in the era of biologicals. HSS J 2006;2:30-41.
11. Paul S, Das AP, Bhattacharjee S. Rheumatoid arthritis: Molecular basis and cures from nature. Int J Pharm Pharm Sci 2015;7:30-9.
12. Rang H, Ritter J, Flower R, Henderson G. Rang and Dale’s Pharmacology. 8th ed. London: Churchill Livingstone; 2015. p. 325-7.
13. Fahmi H, Di Battista JA, Pelletier JP, Mineau F, Ranger P, Martel-Pelletier J, et al. Peroxisome proliferator-activated receptor gamma activators inhibit interleukin-1beta-induced nitric oxide and matrix metalloproteinase 13 production in human chondrocytes. Arthritis Rheum 2001;44:595-607.
14. Clark RB, Bishop-Bailey D, Estrada-Hernandez T, Hla T, Puddington L, Padula SJ, et al. The nuclear receptor PPAR gamma and immunoregulation: PPAR gamma mediates inhibition of helper T cell responses. J Immunol 2000;164:1364-71.
15. Chinetti G, Griglio S, Antonucci M, Torra IP, Delerive P, Majd Z, et al. Activation of proliferator-activated receptors alpha and gamma induces apoptosis of human monocyte-derived macrophages. J Biol Chem 1998;273:25573-80.
16. Harris SG, Phipps RP. The nuclear receptor PPAR gamma is expressed by mouse T lymphocytes and PPAR gamma agonists induce apoptosis. Eur J Immunol 2001;31:1098-105.
17. Chung LY, Schmidt RJ, Andrews AM, Turner TD. A study of hydrogen peroxide generation by, and antioxidant activity of, granuflex (DuoDERM) hydrocolloid granules and some other hydrogel/hydrocolloid wound management materials. Br J Dermatol 1993;129:145-53.
18. Jagetia GC, Baliga MS. The evaluation of nitric oxide scavenging activity of certain Indian medicinal plants in vitro: A preliminary study. J Med Food 2004;7:343-8.
19. Deshpande V, Jadhav VM, Kadam VJ. In-vitro anti-arthritic activity of Abutilon indicum (Linn.) Sweet. J Pharm Res 2009;2:644-5.
20. Gupta SK, Gupta A, Pakash D. In vitro antiarthritic activity of ethanolic extract of Callicarpa macrophylla. Int Res J Pharm 2013;4:160-2.
21. Sakat SS, Juvekar AR, Gambhire MN. In vitro antioxidant and anti-inflammatory activity of methanol extract of Oxalis corniculata Linn. Int J Pharm Pharm Sci 2010;2:146-55.
22. Kyei S, Kofuor G, Boampong J. The efficacy of aqueous and ethanolic leaf extracts of Pistia stratiotes Linn in the management of arthritis and fever. J Med Biomed Sci 2012;1:29-37.
23. Yeom MJ, Lee HC, Kim GH, Lee HJ, Shim I. Antiarthritic effects of Ephedra sinica S TAPF herb-acupuncture: Inhibition of lipopolysaccharide-induced inflammation and adjuvant-induced polyarthritis. J Pharmacol Sci 2006;100:41-50.
24. Mayur P, Kumar A, Maheshwar KK. Establishment of a mechanism of poly herbal formulation for anti-rheumatism. Int J Pharm Pharm Sci 2016;8:91-5.
25. Miller MJ, Sadowsak-Krowicka H, Chotinaruemol S, Kakkis JK, Clark DA. Amelioration of chronic ileitis by nitric oxide synthase inhibition. J Pharmacol Exp Ther 1993;264:11-6.
26. Yermilov V, Rubio J, Becchi M, Friesen MD, Pignatelli B, Ohshima H, et al. Formation of 8-nitroguanine by the reaction of guanine with peroxynitrite in vitro. Carcinogenesis 1995;16:2045-50.
27. Dröge W. Free radicals in the physiological control of cell function. Physiol Rev 2002;82:47-95.
28. Jayaprakasam R, Ravi T. Evaluation of anti-arthritic activity of the root extract of Acalypha indica Linn. Using in vitro techniques. Int J Phytopharm 2013;2:169-73.
29. Anosike CA, Obidoa O, Ezeanyika LU. The anti-inflammatory activity of garden egg (Solanum aethiopicum) on egg albumin-induced oedema and granuloma tissue formation in rats. Asian Pac J Trop Med 2012;5:62-6.
30. Bischoff SC. Quercetin: Potentials in the prevention and therapy of disease. Curr Opin Clin Nutr Metab Care 2008;11:733-40.
31. Besson J, Guilbaud G. The Arthritic Rat as a Model of Clinical Pain? International Congress Series. New York: Excepta Medica Elsevier; 1988. p. 257.
32. Butler SH, Godefroy F, Besson JM, Weil-Fugazza J. A limited arthritic model for chronic pain studies in the rat. Pain 1992;48:73-81.
33. Martínez A, Fernández-Arquero M, Pascual-Salcedo D, Conejero L, Alves H, Balsa A, et al. Primary association of tumor necrosis factor-region genetic markers with susceptibility to rheumatoid arthritis. Arthritis Rheum 2000;43:1366-70.
34. Manivannan P, Jaleel CA, Kishorekumar A, Sankar B, Somasundaram R, Panneerselvam R, et al. Protection of Vigna unguiculata (L.) Walp. Plants from salt stress by paclobutrazol. Colloids Surf B Biointerfaces 2008;61:315-8.
35. Arnett FC, Edworthy SM, Bloch DA, McShane DJ, Fries JF, Cooper NS, et al. The American rheumatism association 1987 revised criteria for the classification of rheumatoid arthritis. Arthritis Rheum 1988;31:315-24.
36. Scott DL, Wolfe F, Huizinga TW. Rheumatoid arthritis. Lancet 2010;376:1094-108.
This work is licensed under a Creative Commons Attribution 4.0 International License.
The publication is licensed under CC By and is open access. Copyright is with author and allowed to retain publishing rights without restrictions.