Konkur Atlarında Egzersiz Öncesi ve Sonrası ADAMTS-4 ve ADAMTS-5 Düzeyleri

Öz

A disintegrin-like and metalloproteinase with thrombospondin motifs (ADAMTS) proteinaz ailesi, spor atlarında lokomotor sistem sağlığının korunması gibi birçok fizyolojik ve fizyopatolojik olayda önemli rol oynamaktadır. Bu çalışmada, yarış atlarında egzersiz öncesi ve sonrasındaki ADAMTS-4 ve ADAMTS-5 düzeylerinin belirlenmesi amaçlanmıştır. Bu amaçla, sağlıklı, 6-15 yaşları arasında, 650-750 kg ağırlığında, farklı cinsiyetlerden, Oldenburg ve Selle Français ırkı yarış atları (n=10) kullanıldı. Atların fiziksel muayeneleri ardından, 50 dakikalık standart egzersiz programı uygulandı. Egzersiz öncesi ve sonrasında ADAMTS-4 ve ADAMTS-5 mRNA ekspresyonlarını ve ELİSA düzeyleri ölçülmek üzere antikoagulansız tüplere kan örnekleri alındı. ADAMTS-4 mRNA ekspresyonu %25 oranında azalmasına rağmen istatistiki olarak anlamlı bulunmadı ve ELİSA düzeyinde de değişim gözlenmedi. ADAMTS-5 mRNA ekspresyonu 3.88 kat arttı (p<0.01) ve ELİSA düzeyi de anlamlı olarak 1.2 kat arttı (p<0.01). Sonuç olarak, serum ADAMTS-5 düzeylerindeki artışın, atlarda eklem hastalıklarının da potansiyel biyolojik belirteçlerinden biri olabileceği ve etkilerini netleştirmek için klinik bulgularla desteklenen daha detaylı araştırmaların yapılması gerektiği kanısına varıldı.

Anahtar Kelimeler

• ADAMTS
• Agrekanaz
• At Egzersiz
• Metalloproteinaz

Pre- and post-exercise ADAMTS-4 and ADAMTS-5 Levels in Concur Horses

Abstract

A disintegrin-like and metalloproteinase with thrombospondin motifs (ADAMTS) proteinase family play an important role in many physiological and physiopathological processes such as the maintenance of locomotor system health in sport horses. In this study, we aimed to determine the changes of ADAMTS-4 and ADAMTS-5 levels in concour horses before and after exercise. The Oldenburg and Selle Français horse-breed types which are healthy, 6-15 years old, around 650-750 kg, and distinct genders were used (n=10). Following the physical examinations, the horses were subjected to 50 minutes of regular exercise program. Blood samples were collected into anticoagulant-free tubes in order to determine ADAMTS-4 and ADAMTS-5 mRNA expression and ELISA levels before and after exercise. There were no differences were observed statistically on ADAMTS-4, neither mRNA expression in spite of 25% downregulated, nor at the ELISA levels. On the other hand, ADAMTS-5 mRNA expression was upregulated 3.88 fold (p<0.01), as well ELISA level significantly increased as 1.2 fold (p<0.01).

Keywords

• ADAMTS
• Aggrecanase
• Equine Exercise
• Metalloproteinase

Kaynakça

1. 1. Murray RC, Dyson SJ, Tranquille C, Adams V. (2006). Association of Type of Sport and Performance Level with Anatomical Site of Orthopaedic Injury Diagnosis. Equine Vet J Suppl. (36): 411-416. doi:10.1111/j.2042-3306.2006.tb05578.x.
2. 2. Lönnell AC, Bröjer J, Nostell K, et al. (2014). Variation in Training Regimens in Professional Showjumping Yards. Equine Vet J. 46(2): 233-238. doi:10.1111/evj.12126.
3. 3. Yamada ALM, Pinheiro M, Marsiglia MF, et al. (2020). Ultrasound and Clinical Findings in the Metacarpophalangeal Joint Assessment of Show Jumping Horses in Training. J Vet Sci. 21.
4. 4. Egenvall A, Tranquille CA, Lonnell AC, et al. (2013). Days-Lost to Training and Competition in Relation to Workload in 263 Elite Show-Jumping Horses in Four European Countries. Prev Vet Med. 112(3-4): 387-400. doi:10.1016/j.prevetmed.2013.09.013.
5. 5. McIlwraith CW, Frisbie DD, Kawcak CE. (2012). The Horse as a Model of Naturally Occurring Osteoarthritis. Bone Joint Res. 1(11): 297-309. doi:10.1302/2046-3758.111.2000132.
6. 6. Chimenti MS, Triggianese P, Conigliaro P, et al. (2015). The Interplay between Inflammation and Metabolism in Rheumatoid Arthritis. Cell Death Dis. 6: e1887. doi:10.1038/cddis.2015.246.
7. 7. Gomes FI, Aragao MG, Barbosa FC, et al. (2016). Inflammatory Cytokines Interleukin-1beta and Tumour Necrosis Factor-Alpha - Novel Biomarkers for the Detection of Periodontal Diseases: A Literature Review. J Oral Maxillofac Res. 7(2): e2. doi:10.5037/jomr.2016.7202.
8. 8. Baker KJ, Houston A, Brint E. (2019). Il-1 Family Members in Cancer; Two Sides to Every Story. Front Immunol. 10: 1197. doi:10.3389/fimmu.2019.01197.

9. 9. Billinghurst RC, Fretz PB, Gordon JR. (1995). Induction of Intra-Articular Tumour Necrosis Factor During Acute Inflammatory Responses in Equine Arthritis. Equine Vet J. 27(3): 208-216.
10. 10. Jouglin M, Robert C, Valette JP, et al. (2000). Metalloproteinases and Tumor Necrosis Factor-Alpha Activities in Synovial Fluids of Horses: Correlation with Articular Cartilage Alterations. Vet Res. 31(5): 507-515. doi:10.1051/vetres:2000136.
11. 11. Kuno K, Kanada N, Nakashima E, et al. (1997). Molecular Cloning of a Gene Encoding a New Type of Metalloproteinase-Disintegrin Family Protein with Thrombospondin Motifs as an Inflammation Associated Gene. J Biol Chem. 272(1): 556-562.
12. 12. Tortorella MD, Burn TC, Pratta MA, et al. (1999). Purification and Cloning of Aggrecanase-1: A Member of the Adamts Family of Proteins. Science. 284(5420): 1664-1666. doi:DOI 10.1126/science.284.5420.1664.
13. 13. Marttinen PH, Raulo SM, Suominen MM, Tulamo RM. (2006). Changes in Mmp-2 and -9 Activity and Mmp-8 Reactivity after Amphotericin B Induced Synovitis and Treatment with Bufexamac. J Vet Med A Physiol Pathol Clin Med. 53(6): 311-318. doi:10.1111/j.1439-0442.2006.00837.x.
14. 14. Majumdar MK, Askew R, Schelling S, et al. (2007). Double-Knockout of Adamts-4 and Adamts-5 in Mice Results in Physiologically Normal Animals and Prevents the Progression of Osteoarthritis. Arthritis Rheum. 56(11): 3670-3674. doi:10.1002/art.23027.
15. 15. Kandir S. (2020). Adamts Proteases: Potential Biomarkers and Novel Therapeutic Targets for Cartilage Health. In Equine Science. (Ruthland C ed). pp. 1-15. IntechOpen, London. doi:10.5772/intechopen.93046
16. 16. Abbaszade I, Liu RQ, Yang F, et al. (1999). Cloning and Characterization of Adamts11, an Aggrecanase from the Adamts Family. J Biol Chem. 274(33): 23443-23450. doi:10.1074/jbc.274.33.23443.
17. 17. Tortorella MD, Burn TC, Pratta MA, et al. (1999). Purification and Cloning of Aggrecanase-1: A Member of the Adamts Family of Proteins. Science. 284(5420): 1664-1666. doi:10.1126/science.284.5420.1664.
18. 18. Pawlak E, Wang L, Johnson PJ, et al. (2012). Distribution and Processing of a Disintegrin and Metalloproteinase with Thrombospondin Motifs-4, Aggrecan, Versican, and Hyaluronan in Equine Digital Laminae. Am J Vet Res. 73(7): 1035-1046. doi:10.2460/ajvr.73.7.1035.
19. 19. Peffers MJ, Thornton DJ, Clegg PD. (2016). Characterization of Neopeptides in Equine Articular Cartilage Degradation. J Orthop Res. 34(1): 106-120. doi:10.1002/jor.22963.
20. 20. Moon J-W, Ahn K, Bae J-H, et al. (2012). Mrna Sequence Analysis and Quantitative Expression of the Adamts4 Gene in the Thoroughbred Horse. Genes & Genomics. 34(4): 441-445. doi:10.1007/s13258-011-0204-0.
21. 21. Huang K, Wu LD. (2008). Aggrecanase and Aggrecan Degradation in Osteoarthritis: A Review. J Int Med Res. 36(6): 1149-1160. doi:Doi 10.1177/147323000803600601.
22. 22. Coyne MJ, Cousin H, Loftus JP, et al. (2009). Cloning and Expression of Adam-Related Metalloproteases in Equine Laminitis. Vet Immunol Immunop. 129(3-4): 231-241. doi:10.1016/j.vetimm.2008.11.022.
23. 23. Velasco J, Li J, DiPietro L, et al. (2011). Adamts5 Deletion Blocks Murine Dermal Repair through Cd44-Mediated Aggrecan Accumulation and Modulation of Transforming Growth Factor Beta1 (Tgfbeta1) Signaling. J Biol Chem. 286(29): 26016-26027. doi:10.1074/jbc.M110.208694.
24. 24. Will H, Dettloff M, Bendzko P, Sveshnikov P. (2005). A Quantitative Assay for Aggrecanase Activity. J Biomol Tech. 16(4): 459-472.
25. 25. Dutto DJ, Hoyt DF, Clayton HM, Cogger EA, Wickler SJ. (2004). Moments and Power Generated by the Horse (Equus Caballus) Hind Limb During Jumping. J Exp Biol. 207(Pt 4): 667-674. doi:10.1242/jeb.00808.
26. 26. Meershoek LS, Schamhardt HC, Roepstorff L, Johnston C. (2001). Forelimb Tendon Loading During Jump Landings and the Influence of Fence Height. Equine Vet J Suppl. (33): 6-10.
27. 27. Krumrych W, Gołda R, Gołyński M, Markiewicz H, Buzała M. (2018). Effect of Physical Exercise on Cortisol Concentration and Neutrophil Oxygen Metabolism in Peripheral Blood of Horses. 18(1): 53. doi:https://doi.org/10.1515/aoas-2017-0019.
28. 28. Woods J, Lu Q, Ceddia MA, Lowder T. (2000). Special Feature for the Olympics: Effects of Exercise on the Immune System: Exercise-Induced Modulation of Macrophage Function. Immunol Cell Biol. 78(5): 545-553. doi:10.1111/j.1440-1711.2000.t01-9-.x.
29. 29. Gomez-Aristizabal A, Gandhi R, Mahomed NN, Marshall KW, Viswanathan S. (2019). Synovial Fluid Monocyte/Macrophage Subsets and Their Correlation to Patient-Reported Outcomes in Osteoarthritic Patients: A Cohort Study. Arthritis Res Ther. 21(1): 26. doi:10.1186/s13075-018-1798-2.
30. 30. Tu J, Hong W, Zhang P, et al. (2018). Ontology and Function of Fibroblast-Like and Macrophage-Like Synoviocytes: How Do They Talk to Each Other and Can They Be Targeted for Rheumatoid Arthritis Therapy? Front Immunol. 9: 1467. doi:10.3389/fimmu.2018.01467.
31. 31. Nees TA, Rosshirt N, Zhang JA, et al. (2019). Synovial Cytokines Significantly Correlate with Osteoarthritis-Related Knee Pain and Disability: Inflammatory Mediators of Potential Clinical Relevance. J Clin Med. 8(9). doi:10.3390/jcm8091343.
32. 32. Bondeson J, Wainwright S, Hughes C, Caterson B. (2008). The Regulation of the Adamts4 and Adamts5 Aggrecanases in Osteoarthritis: A Review. Clin Exp Rheumatol. 26(1): 139-145.
33. 33. Vonk AG, Netea MG, Denecker NE, et al. (1998). Modulation of the Pro- and Anti-Inflammatory Cytokine Balance by Amphotericin B. J Antimicrob Chemother. 42(4): 469-474.
34. 34. Tokuda Y, Tsuji M, Yamazaki M, et al. (1993). Augmentation of Murine Tumor Necrosis Factor Production by Amphotericin B in Vitro and in Vivo. Antimicrob Agents Chemother. 37(10): 2228-2230.
35. 35. Mokbel AN, El Tookhy OS, Shamaa AA, et al. (2011). Homing and Reparative Effect of Intra-Articular Injection of Autologus Mesenchymal Stem Cells in Osteoarthritic Animal Model. BMC Musculoskelet Disord. 12: 259. doi:10.1186/1471-2474-12-259.
36. 36. Ma TW, Li Y, Wang GY, et al. (2017). Changes in Synovial Fluid Biomarkers after Experimental Equine Osteoarthritis. J Vet Res. 61(4): 503-508. doi:10.1515/jvetres-2017-0056.
37. 37. McGivney BA, Eivers SS, MacHugh DE, et al. (2009). Transcriptional Adaptations Following Exercise in Thoroughbred Horse Skeletal Muscle Highlights Molecular Mechanisms That Lead to Muscle Hypertrophy. Bmc Genomics. 10. doi:10.1186/1471-2164-10-638. 38. Zhou X, Jiang L, Zhang Y, et al. (2019). Genetic Variation of Aggrecanase-2 (Adamts5) in Susceptibility to Osteoarthritis. Braz J Med Biol Res. 52(2): e8109-e8109. doi:10.1590/1414-431X20188109.
38. 39. Canbek U, Imerci A, Kara M, et al. (2016). Polymorphisms in Adamts4 and Adamts5 Are Not Linked to Susceptibility to Knee Osteoarthritis in the Turkish Population. Genetics and molecular research : GMR. 15(3). doi:10.4238/gmr.15038264.