Comparison of intra-articular injection of Naproxen and glutathione in treatment of knee rheumatoid arthritis in a male rabbit model

I. H. Jirjees; Z. K. Mahmood; H. M. Ehymayed; F. K. Tawfeeq

doi:10.15421/0225152

I. H. Jirjees University of Mosul
Z. K. Mahmood Northern Technical University
H. M. Ehymayed Al-Noor University College
F. K. Tawfeeq University of Mosul

Keywords: rheumatoid arthritis, intra-articular injection, glutathione, Naproxen.

Abstract

Rheumatoid arthritis (RA) is a chronic progressive inflammatory joint disorder. Targeting drug delivery spare s adverse e f fects and optimize s therapy. This study sought to compare the hematological and immunological impacts of intra-articular N a proxen sodium and glutathione (GSH) in an arthritis rabbit model. Twelve adult male rabbits (2.5 – 3.5 kg weight), were placed in separate cages under standard conditions and divided into 4 groups (3 each). The p ositive control group (PCG) was given intra-articular (IA) bovine serum albumin (BSA), the negative control group (NCG) was given IA normal saline, the GSH group was given IA BSA with GSH for three days, and the N aproxen group was given IA BSA with N aproxen for three days. Blood samples were collected from the lateral saphenous vein and marginal ear vein at day seven post treatment with BSA into plain tube s to separate serum for the rheumatoid factor (RF) test. The second collection of blood fraction was used for blood indices, including RBCs (red blood cells), Hb (hemoglobin), HCT (hematocrit), MCV (mean corpuscular volume), MCH (mean corpu s cular hemoglobin), MCHC (mean corpuscular hemoglobin concentration), RDWCV (red cell distribution width – coefficient of variation). The N aproxen group showed the highest RBCs (6.4 ± 0.2), Hb (14.2 ± 0. 4 ), and MCHC (33.6 ± 0.5), while also demonstrat ing elevated MCH (22.3 ± 0.6) and maintaining HCT levels (42. 1 ± 1.0). The GSH group demonstrated the lowest levels of these parameters. Regarding WBCs, the GSH group showed the highest value in WBCs (10.5 ± 1.64), compared to the positive control ( P = 0.003), negative control ( P = 0.021), and N aproxen groups ( P < 0.001). The N aproxen group demonstrated the lowest total WBC count (5.7 ± 1 . 0 ) and markedly reduced lymphocyte percentage (55.9 ± 6. 1 ) compared to all other trea t ment groups. The PLT count revealed marked variations in the four treatment groups, with the positive control group demo n strat ing significantly lower PLT versus all other groups. The positive control group reported significantly elevated RF (24.0 ± 13.5) levels compared to other treatment groups ( P < 0.001 for negative control and glutathione, P = 0.002 for N aproxen). The results confirmed significant improvement in WBC count in the glutathione group compared to the N aproxen group. This study concluded that an intra-articular glutathione injection may alleviate arthritis . Yet, further investigation is required to better unde r stand the role of glutathione role in the healing course of arthritis.

References

Bilski, R., & Nuszkiewicz, J. (2025). Antioxidant therapies as emerging adjuncts in rheumatoid arthritis: Targeting oxidative stress to enhance treatment outcomes. International Journal of Molecular Sciences, 26(7), 2873.

Brown, P., Pratt, A. G., & Hyrich, K. L. (2024). Therapeutic advances in rheumatoid arthritis. British Medical Journal, 384, e070856.

Butoescu, N., Jordan, O., & Doelker, E. (2009). Intra-articular drug delivery systems for the treatment of rheumatic diseases: A review of the factors influencing their performance. European Journal of Pharmaceutics and Biopharmaceutics, 73(2), 205–218.

Ceuppens, J. L., Robaeys, G., Verdickt, W., Vertessen, S., Deckmyn, H., & Dequeker, J. (1986). Immunomodulatory effects of treatment with Naproxen in patients with rheumatic disease. Arthritis and Rheumatism, 29(3), 305–311.

Cicala, C., Ianaro, A., Fiorucci, S., Calignano, A., Bucci, M., Gerli, R., Santucci, L., Wallace, J. L., & Cirino, G. (2000). NO-naproxen modulates inflammation, nociception and downregulates T cell response in rat Freund’s adjuvant arthritis. British Journal of Pharmacology, 130(6), 1399–1405.

Conran, C., Kolfenbach, J., Kuhn, K., Striebich, C., & Moreland, L. (2023). A review of difficult-to-treat rheumatoid arthritis: Definition, clinical presentation, and management. Current Rheumatology Reports, 25(12), 285–294.

Darwish, A. A., & Eldakroury, M. F. (2020). The effect of some anti-inflammatory drugs on some clinicopathological parameters in Barki sheep. Journal of Animal Health and Production, 8(3), 93–100.

De La Cruz, J. P., Reyes, J. J., Ruiz-Moreno, M. I., Lopez-Villodres, J. A., Jebrouni, N., & Gonzalez-Correa, J. A. (2010). Differences in the in vitro antiplatelet effect of Dexibuprofen, Ibuprofen, and Flurbiprofen in human blood. Anesthesia and Analgesia, 111(6), 1341–1346.

Diotallevi, M., Checconi, P., Palamara, A. T., Celestino, I., Coppo, L., Holmgren, A., Abbas, K., Peyrot, F., Mengozzi, M., & Ghezzi, P. (2017). Glutathione fine-tunes the innate immune response toward antiviral pathways in a macrophage cell line independently of its antioxidant properties. Frontiers in Immunology, 8, 1239.

Dobre, I. R., Alexandru, D. M., & Crivineanu, M. (2019). Study on changes in haematological parameters following administration of NSAIDs in dogs. Scientific Works, C Series, Veterinary Medicine, 65(1), 37–41.

Donahue, K. E., Gartlehner, G., Jonas, D. E., Lux, L. J., Thieda, P., Jonas, B. L., Hansen, R. A., Morgan, L. C., & Lohr, K. N. (2008). Systematic review: Comparative effectiveness and harms of disease-modifying medications for rheumatoid arthritis. Annals of Internal Medicine, 148(2), 124–134.

Evans, C. H. (2005). Novel biological approaches to the intra-articular treatment of osteoarthritis. BioDrugs, 19(6), 355–362.

Finckh, A., Gilbert, B., Hodkinson, B., Bae, S.-C., Thomas, R., Deane, K. D., Alpizar-Rodriguez, D., & Lauper, K. (2022). Global epidemiology of rheumatoid arthritis. Nature Reviews Rheumatology, 18, 591–602.

Fonseca, L. J. S. D., Nunes-Souza, V., Goulart, M. O. F., & Rabelo, L. A. (2019). Oxidative stress in rheumatoid arthritis: What the future might hold regarding novel biomarkers and add-on therapies. Oxidative Medicine and Cellular Longevity, 2019, 7536805.

Gomaa, S. (2018). Adverse effects induced by Diclofenac, Ibuprofen, and Paracetamol toxicity on immunological and biochemical parameters in Swiss albino mice. The Journal of Basic and Applied Zoology, 79, 5.

Jones, I. A., Togashi, R., Wilson, M. L., Heckmann, N., & Vangsness, C. T. (2019). Intra-articular treatment options for knee osteoarthritis. Nature Reviews Rheumatology, 15(2), 77–90.

Kao, D. S., Zhang, S. W., & Vap, A. R. (2022). A systematic review on the effect of common medications on platelet count and function: Which medications should be stopped before getting a platelet-rich plasma injection? Orthopaedic Journal of Sports Medicine, 10(4), 23259671221088820.

Kaur, G., Sharma, A., & Bhatnagar, A. (2021). Role of oxidative stress in pathophysiology of rheumatoid arthritis: Insights into NRF2-KEAP1 signalling. Autoimmunity, 54(7), 385–397.

Khanna, N., Kumar, A., & Pawar, S. V. (2021). A review on rheumatoid arthritis interventions and current developments. Current Drug Targets, 22(4), 463–483.

Lei, Y., Wang, K., Deng, L., Chen, Y., Nice, E. C., & Huang, C. (2015). Redox regulation of inflammation: Old elements, a new story. Medicinal Research Reviews, 35(2), 306–340.

Mannava, S., Whitney, K. E., Kennedy, M. I., King, J., Dornan, G. J., Klett, K., Chahla, J., Evans, T. A., Huard, J., & LaPrade, R. F. (2019). The influence of naproxen on biological factors in leukocyte-rich platelet-rich plasma: A prospective comparative study. Arthroscopy, 35(1), 201–210.

Matloob, R., Althanoon, Z., Algburi, S., Salih, M., & Merkhan, M. (2024). Update on the use of methotrexate in the management of rheumatoid arthritis. Georgian Medical News, 347, 28–33.

Ogidi, O. I., Ogoun, T. R., Njoku, C. O., Charles, E. E., Amgbare, E. B., & Omotehinse, E. T. (2020). Toxicity studies on the effects of non-steroidal anti-inflammatory drugs in Wistar albino rats. Elixir Pharmacy, 149, 55010–55014.

Parolini, M. (2020). Toxicity of the non-steroidal anti-inflammatory drugs (NSAIDs) acetylsalicylic acid, Paracetamol, Diclofenac, Ibuprofen and Naproxen towards freshwater invertebrates: A review. Science of the Total Environment, 740, 140043.

Prasad, P., Verma, S., Surbhi, Ganguly, N. K., Chaturvedi, V., & Mittal, S. A. (2023). Rheumatoid arthritis: Advances in treatment strategies. Molecular and Cellular Biochemistry, 478, 69–88.

Pretorius, E., Akeredolu, O.-O., Soma, P., & Kell, D. B. (2017). Major involvement of bacterial components in rheumatoid arthritis and its accompanying oxidative stress, systemic inflammation and hypercoagulability. Experimental Biology and Medicine, 242(4), 355–373.

Rahman, I. (2000). Regulation of nuclear factor-κB, activator protein-1, and glutathione levels by tumor necrosis factor-α and dexamethasone in alveolar epithelial cells. Biochemical Pharmacology, 60(8), 1041–1049.

Razi, M. T., Javed, I., Choudry, M. Z., Khan, M. T., & Mukhtar, N. (2014). Effect of Ketoprofen on lactic dehydrogenasefrom human platelets. Advances in Clinical and Experimental Medicine, 23(3), 377–380.

Ribas, J. L. C., Zampronio, A. R., & Silva De Assis, H. C. (2015). Effects of trophic exposure to Diclofenac and Dexamethasone on hematological parameters and immune response in freshwater fish. Environmental Toxicology and Chemistry, 35(4), 975–982.

Saper, C. B., Romanovsky, A. A., & Scammell, T. E. (2012). Neural circuitry engaged by prostaglandins during the sickness syndrome. Nature Neuroscience, 15(8), 1088–1095.

Saravanan, M., Devi, K. U., Malarvizhi, A., & Ramesh, M. (2012). Effects of Ibuprofen on hematological, biochemical and enzymological parameters of blood in an Indian major carp, Cirrhinus mrigala. Environmental Toxicology and Pharmacology, 34(1), 14–22.

Silvagno, F., Vernone, A., & Pescarmona, G. P. (2020). The role of glutathione in protecting against the severe inflammatory response triggered by COVID-19. Antioxidants, 9(7), 624.

Smolen, J. S., Landewé, R. B. M., Bijlsma, J. W. J., Burmester, G. R., Dougados, M., Kerschbaumer, A., McInnes, I. B., Sepriano, A., Van Vollenhoven, R. F., De Wit, M., Aletaha, D., Aringer, M., Askling, J., Balsa, A., Boers, M., Den Broeder, A. A., Buch, M. H., Buttgereit, F., Caporali, R., … Van Der Heijde, D. (2020). EULAR recommendations for the management of rheumatoid arthritis with synthetic and biological disease-modifying antirheumatic drugs: 2019 update. Annals of the Rheumatic Diseases, 79(6), 685–699.

Zamudio-Cuevas, Y., Martínez-Flores, K., Martínez-Nava, G. A., Clavijo-Cornejo, D., Fernández-Torres, J., & Sánchez-Sánchez, R. (2022). Rheumatoid arthritis and oxidative stress, a review of a decade. Cellular and Molecular Biology, 68(6), 174–184.