Ukr.Biochem.J. 2021; Volume 93, Issue 5, Sep-Oct, pp. 63-71


Metallothioneins involment in the pathogenesis of synovial tissue inflammation in rats with acute gonarthritis

T. R. Matskiv1,2, D. V. Lytkin3, S. K. Shebeko3, V. V. Khoma2,
V. V. Martyniuk2, L. L. Gnatyshyna1,2, O. B. Stoliar2*

1I. Horbachevsky Ternopil National Medical University, Department of General Chemistry, Ternopil, Ukraine;
2Ternopil Volodymyr Hnatiuk National Pedagogical University, Department of Chemistry and Methods its Teaching, Ternopil, Ukraine;
3National University of Pharmacy, Educational and Scientific Institute of Applied Pharmacy, Kharkiv, Ukraine;

Received: 02 June 2021; Accepted: 22 September 2021

Zinc (Zn) is involving in the suppressing of inflammation. However, its functionality in the knee joint under the gonarthritis (GA) is not elucidated. The aim of this study was to investigate the participation of Zn-buffering and stress responsive proteins metallothioneins (MTs) in the pathogenesis of the synovial tissues under the experimental acute GA. The inflammation was induced in rats by intra-articular administration of carrageenan. The concentrations of MTs total protein (MTSH), Zn-bound protein (Zn-MTs), total Zn concentration in the tissue, the indexes of oxidative stress and cholinesterase activity were determined. The level of sialic acids was indicated in the blood serum. The enhancing of sialic acids concentration by 42% and cholinesterase depletion confirmed the pathology. In the animals with GA, total level of Zn in the tissue was correspondent to control. However, the MTSH and Zn-MT levels were elevated (by 79 and 46% respectively). This disproportionate rate can be due to partial oxidation of thiols. The superoxide dismutase activity was elevated, radical scavenging activity and protein carbonylation were correspondent to control, but the levels of catalase, glutathione–S-transferase and glutathione  were decreased by 28-44%, and lipid peroxidation (TBARS) was increased by 59% compared to control group. Principal Component Analysis confirmed the strong interrelations between MTs and peroxide-related oxidative stress indexes. This preliminary study provides the basis for the understanding of the reason for Zn imbalance in the acute GA as the result of the impairment of thiol redox balance and proposes these biomarkers for the evaluation of knee joint pathologies.

Keywords: , , ,


  1. Rice JM, Zweifach A, Lynes MA. Metallothionein regulates intracellular zinc signaling during CD4(+) T cell activation. BMC Immunol. 2016;17(1):13. PubMed, PubMedCentral, CrossRef
  2. Krężel A, Maret W. The biological inorganic chemistry of zinc ions. Arch Biochem Biophys. 2016;611:3-19. PubMed, PubMedCentral, CrossRef
  3. Frangos T, Maret  W. Zinc and Cadmium in the Aetiology and Pathogenesis of Osteoarthritis and Rheumatoid Arthritis. Nutrients. 2020;13(1):53. PubMed, PubMedCentral, CrossRef
  4. Sun J, Li L, Li L, Ding L, Liu X, Chen X, Zhang J, Qi X, Du J, Huang Z. Metallothionein-1 suppresses rheumatoid arthritis pathogenesis by shifting the Th17/Treg balance. Eur J Immunol. 2018;48(9):1550-1562. PubMed, CrossRef
  5. Lespasio MJ, Piuzzi NS, Husni ME, Muschler GF, Guarino A, Mont MA. Knee Osteoarthritis: A Primer. Perm J. 2017;21:16-183. PubMed, PubMedCentral, CrossRef
  6. Sur B, Kang S, Kim M, Oh S. Inhibition of Carrageenan/Kaolin-Induced Arthritis in Rats and of Inflammatory Cytokine Expressions in Human IL-1β-Stimulated Fibroblast-like Synoviocytes by a Benzylideneacetophenone Derivative. Inflammation. 2019;42(3):928-936. PubMed, PubMedCentral, CrossRef
  7. Viarengo A, Ponzano E, Dondero F, Fabbri R. A simple spectrophotometric method for metallothionein evaluation in marine organisms: an application to Mediterranean and Antarctic Molluscs. Mar Environ Res. 1997;44(1):69-84.  CrossRef
  8. Suzuki KT.  Purification of vertebrate metallothioneins. Methods Enzymol. 1991;205:252-263. PubMed, CrossRef
  9. Wang J, Niu Y, Zhang C, Chen Y. A micro-plate colorimetric assay for rapid determination of trace zinc in animal feed, pet food and drinking water by ion masking and statistical partitioning correction. Food Chem. 2018;245:337-345. PubMed, CrossRef
  10. Re R, Pellegrini N, Proteggente A, Pannala A, Yang M, Rice-Evans C. Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radic Biol Med. 1999;26(9-10):1231-1237. PubMed, CrossRef
  11. Beauchamp C, Fridovich I. Superoxide dismutase: improved assays and an assay applicable to acrylamide gels. Anal Biochem. 1971;44(1):276-287. PubMed, CrossRef
  12. Aebi H, Bergmeyer HU. Catalase. Methods Enzym Anal. 1974;2:673-677.  CrossRef
  13. Habig WH, Pabst MJ, Jakoby WB. Glutathione S-transferases. The first enzymatic step in mercapturic acid formation. J Biol Chem. 1974;249(22):7130-7139. PubMed, CrossRef
  14. Griffith OW. Determination of glutathione and glutathione disulfide using glutathione reductase and 2-vinylpyridine. Anal Biochem. 1980;106(1):207-212. PubMed, PubMedCentral, CrossRef
  15. Ohkawa H, Ohishi N, Yagi K. Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem. 1979;95(2):351-358. PubMed, CrossRef
  16. Reznick AZ, Packer L. Oxidative damage to proteins: spectrophotometric method for carbonyl assay. Methods Enzymol. 1994;233:357-363. PubMed, CrossRef
  17. Ellman GL, Courtney KD, Andres VJr, Featherstone RM. A new and rapid colorimetric determination of acetylcholinesterase activity. Biochem Pharmacol. 1961;7:88-95. PubMed, CrossRef
  18.  Khoma V, Gnatyshyna L, Martinyuk V, Mackiv T, Mishchenko L, Manusadžianas L, Stoliar O. Common and particular biochemical responses of Unio tumidus to herbicide, pharmaceuticals and their combined exposure with heating. Ecotoxicol Environ Saf. 2021;208:111695. PubMed, CrossRef
  19. Lynes MA, Zaffut K, Unfricht DW, Marusov G, Samson JS, Yin X. The physiological roles of extracellular metallothionein. Exp Biol Med (Maywood). 2006;231(9):1548-1554. PubMed, CrossRef
  20. Grider A, Cousins RJ. Role of metallothionein in copper and zinc metabolism: special reference to inflammatory conditions. In: Milanino R, Rainsford KD, Velo GP (Eds.). Copper and Zinc in Inflammation. Inflammation and Drug Therapy Series. Dordrecht, Springer 1989;4:21-32.  CrossRef
  21. Hwang J, Jin J, Jeon S, Moon SH, Park MY, Yum DY, Kim JH, Kang JE, Park MH, Kim EJ, Pan JG, Kwon O, Oh GT.  SOD1 suppresses pro-inflammatory immune responses by protecting against oxidative stress in colitis. Redox Biol. 2020;37:101760. PubMed, PubMedCentral, CrossRef
  22. Scott JL, Gabrielides C, Davidson RK, Swingler TE, Clark IM, Wallis GA, Boot-Handford RP, Kirkwood TBL, Taylor RW, Young DA. Superoxide dismutase downregulation in osteoarthritis progression and end-stage disease. Ann Rheum Dis. 2010;69(8):1502-1510. PubMed, PubMedCentral, CrossRef
  23. García-González A, Gaxiola-Robles R, Zenteno-Savín T. Oxidative stress in patients with rheumatoid arthritis. Rev Invest Clin. 2015;67(1):46-53. PubMed
  24. Dunning DL, Holmes J, Gathercole SE. Does working memory training lead to generalized improvements in children with low working memory? A randomized controlled trial. Dev Sci. 2013;16(6):915-925. PubMed, PubMedCentral, CrossRef
  25. Dröge W. Free radicals in the physiological control of cell function. Physiol Rev. 2002;82(1):47-95. PubMed, CrossRef
  26. Rudyk O, Eaton P. Biochemical methods for monitoring protein thiol redox states in biological systems. Redox Biol. 2014;2:803-813. PubMed, PubMedCentral, CrossRef
  27. Wang Y, Weisenhorn E, MacDiarmid CW, Andreini C, Bucci M, Taggart J, Banci L, Russell J, Coon JJ, Eide DJ. The cellular economy of the Saccharomyces cerevisiae zinc proteome. Metallomics. 2018;10(12):1755-1776. PubMed, PubMedCentral, CrossRef
  28. Hoover DB. Cholinergic modulation of the immune system presents new approaches for treating inflammation. Pharmacol Ther. 2017;179:1-16. PubMed, PubMedCentral, CrossRef
  29. Itoh Y. Metalloproteinases in Rheumatoid Arthritis: Potential Therapeutic Targets to Improve Current Therapies. Prog Mol Biol Transl Sci. 2017;148:327-338. PubMed, CrossRef
  30. Won Y, Shin Y, Chun CH, Cho Y, Ha CW, Kim JH, Chun JS. Pleiotropic roles of metallothioneins as regulators of chondrocyte apoptosis and catabolic and anabolic pathways during osteoarthritis pathogenesis. Ann Rheum Dis. 2016;75(11):2045-2052. PubMed, PubMedCentral, CrossRef

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