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Action of three bioavailable antioxidants in orbital fibroblasts from patients with Graves’ orbitopathy (GO): a new frontier for GO treatment?

Journal of Endocrinological Investigation volume 41, pages 193–201 (2018)Cite this article

Abstract

Objective

Oxidative stress is involved in the pathogenesis of Graves’ orbitopathy (GO) and an antioxidant approach has been advocated for GO treatment. Here, we investigated the action of three antioxidants in orbital fibroblasts, namely, vitamin C, N-acetyl-l-cysteine, and melatonin.

Methods

Primary cultures of orbital fibroblasts from six GO patients and six control subjects were established. Cells were treated with H2O2 to induce oxidative stress. Cell vitality assays were performed to determine the non-cytotoxic dose of each antioxidant. The following assays were performed: glutathione disulfide (GSSG), as a measure of oxidative stress, cell proliferation, hyaluronic acid (HA), TNFα, IFNγ, and IL1β.

Results

H2O2 induced oxidative stress (augmented GSSG), increased cell proliferation as well as cytokine release, but did not affect HA release. All of the three antioxidant substances reduced H2O2-dependent oxidative stress. Vitamin C reduced proliferation in GO, but not in control fibroblasts. N-acetyl-l-cysteine reduced proliferation and IFNγ in GO, and HA and IL1β in both GO and control fibroblasts. Melatonin reduced IL1β and HA in GO and control fibroblasts, and IFNγ only in GO fibroblasts.

Conclusions

Our study provides evidence in support of an antioxidant role of vitamin C, N-acetyl-l-cysteine and melatonin in orbital fibroblasts. Some of the effects of these compounds are exclusive to GO fibroblasts, whereas some other are observed also in control fibroblasts. Our observations provide a basis for a possible clinical use of these substances in patients with GO.

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References

  1. Bahn RS (2010) Graves’ ophthalmopathy. N Engl J Med 362:726–738

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  2. Bartalena L (2013) Diagnosis and management of Graves disease: a global overview. Nat Rev Endocrinol 9:724–734

    CAS  Article  PubMed  Google Scholar 

  3. Piantanida E, Tanda ML, Lai A, Sassi L, Bartalena L (2013) Prevalence and natural history of Graves’ orbitopathy in the XXI century. J Endocrinol Invest 36:444–449

    CAS  PubMed  Google Scholar 

  4. Leo M, Menconi F, Rocchi R, Latrofa F, Sisti E, Profilo MA, Mazzi B, Albano E, Nardi M, Vitti P, Marcocci C, Marinò M (2015) Role of the underlying thyroid disease on the phenotype of Graves’ orbitopathy in a tertiary referral center. Thyroid 25:347–351

    Article  PubMed  Google Scholar 

  5. Sisti E, Menconi F, Leo M, Profilo MA, Mautone T, Mazzi B, Rocchi R, Latrofa F, Nardi M, Vitti P, Marcocci C, Marinò M (2015) Long-term outcome of Graves’ orbitopathy following high-dose intravenous glucocorticoids and orbital radiotherapy. J Endocrinol Invest 38:661–668

    CAS  Article  PubMed  Google Scholar 

  6. Menconi F, Leo M, Sabini E, Mautone T, Nardi M, Sainato A, Sellari-Franceschini S, Vitti P, Marcocci C, Marinò M (2016) Natural history of Graves’ orbitopathy after treatment. Endocrine (in press)

  7. Bartalena L, Veronesi G, Krassas GE, Wiersinga WM, Marcocci C, Marinò M, Salvi M, Daumerie C, Bournaud C, Stahl M, Sassi L, Azzolini C, Boboridis KG, Mourits MP, Soeters MR, Baldeschi L, Nardi M, Currò N, Boschi A, Bernard M, von Arx G, Perros P, Kahaly GJ, European Group on Graves’ Orbitopathy (EUGOGO) (2017) Does early response to intravenous glucocorticoids predict the final outcome in patients with moderate-to-severe and active Graves’ orbitopathy? J Endocrinol Invest 40:547–553

    CAS  Article  PubMed  Google Scholar 

  8. Bartalena L, Baldeschi L, Boboridis K, Eckstein A, Kahaly GJ, Marcocci C, Perros P, Salvi M, Wiersinga WM, European Group on Graves’ Orbitopathy (EUGOGO) (2016) The 2016 European Thyroid Association/European Group on Graves’ orbitopathy guidelines for the management of graves’ orbitopathy. Eur Thyroid J 5:9–26

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  9. Menconi F, Profilo MA, Leo M, Sisti E, Altea MA, Rocchi R, Latrofa F, Nardi M, Vitti P, Marcocci C, Marinò M (2014) Spontaneous improvement of untreated mild Graves’ ophthalmopathy: Rundle’s curve revisited. Thyroid 24:60–66

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  10. Anagnostis P, Boboridis K, Adamidou F, Kita M (2017) Natural course of mild Graves’ orbitopathy: is it a chronic remitting or a transient disease? J Endocrinol Invest 40:257–261

    CAS  Article  PubMed  Google Scholar 

  11. Terwee CB, Gerding MN, Dekker FW, Prummel MF, Wiersinga WM (1998) Development of a disease specific quality of life questionnaire for patients with Graves’ ophthalmopathy: the GO-QOL. Br J Ophthalmol 82:773–779

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  12. Zang S, Ponto KA, Kahaly GJ (2011) Clinical review: intravenous glucocorticoids for Graves’ orbitopathy: efficacy and morbidity. J Clin Endocrinol Metab 96:320–332

    CAS  Article  PubMed  Google Scholar 

  13. Sisti E, Coco B, Menconi F, Leo M, Rocchi R, Latrofa F, Profilo MA, Mazzi B, Albano E, Vitti P, Marcocci C, Brunetto M, Marinò M (2015) Intravenous glucocorticoid therapy for Graves’ ophthalmopathy and acute liver damage: an epidemiological study. Eur J Endocrinol 172:269–276

    CAS  Article  PubMed  Google Scholar 

  14. Sisti E, Coco B, Menconi F, Leo M, Rocchi R, Latrofa F, Profilo MA, Mazzi B, Vitti P, Marcocci C, Brunetto M, Marinò M (2015) Age and dose are major risk factors for liver damage associated with intravenous glucocorticoid pulse therapy for Graves’ orbitopathy. Thyroid 25:846–850

    CAS  Article  PubMed  Google Scholar 

  15. Moleti M, Giuffrida G, Sturniolo G, Squadrito G, Campennì A, Morelli S, Puxeddu E, Sisti E, Trimarchi F, Vermiglio F, Marinò M (2016) Acute liver damage following intravenous glucocorticoid treatment for Graves’ ophthalmopathy. Endocrine 54:259–268

    CAS  Article  PubMed  Google Scholar 

  16. Sabini E, Sisti E, Coco B, Leo M, Ionni I, Menconi F, Profilo MA, Mazzi B, Rocchi R, Latrofa F, Vitti P, Brunetto M, Marcocci C, Marinò M (2016) Statins are not a risk factor for liver damage associated with intravenous glucocorticoid pulse therapy for Graves’ orbitopathy. J Endocrinol Invest 39:1323–1327

    CAS  Article  PubMed  Google Scholar 

  17. Burch HB, Lahiri S, Bahn RS, Barnes S (1997) Superoxide radical production stimulates retroocular fibroblast proliferation in Graves’ ophthalmopathy. Exp Eye Res 65:311–316

    CAS  Article  PubMed  Google Scholar 

  18. Heufelder AE, Wenzel BE, Bahn RS (1992) Methimazole and propylthiouracil inhibit the oxygen free radical-induced expression of a 72 kilodalton heat shock protein in Graves’ retroocular fibroblasts. J Clin Endocrinol Metab 74:737–742

    CAS  Article  PubMed  Google Scholar 

  19. Lu R, Wang P, Wartofsky L, Sutton BD, Zweier JL, Bahn RS, Garrity J, Burman KD (1999) Oxygen free radicals in interleukin-1beta-induced glycosaminoglycan production by retro-ocular fibroblasts from normal subjects and Graves’ ophthalmopathy patients. Thyroid 9:297–303

    CAS  Article  PubMed  Google Scholar 

  20. Marcocci C, Kahaly GJ, Krassas GE, Bartalena L, Prummel M, Stahl M, Altea MA, Nardi M, Pitz S, Boboridis K, Sivelli P, von Arx G, Mourits MP, Baldeschi L, Bencivelli W, Wiersinga W (2011) Selenium and the course of mild Graves’ orbitopathy. N Engl J Med 364:1920–1931

    CAS  Article  PubMed  Google Scholar 

  21. Leo M, Bartalena L, Rotondo Dottore G, Piantanida E, Premoli P, Ionni I, Di Cera M, Masiello E, Sassi L, Tanda ML, Latrofa F, Vitti P, Marcocci C, Marinò M (2017) Effects of selenium on short-term control of hyperthyroidism due to Graves’ disease treated with methimazole: results of a randomized clinical trial. J Endocrinol Invest 40:281–287

    CAS  Article  PubMed  Google Scholar 

  22. Esposito D, Rotondi M, Accardo G, Vallone G, Conzo G, Docimo G, Selvaggi F, Cappelli C, Chiovato L, Giugliano D, Pasquali D (2017) Influence of short-term selenium supplementation on the natural course of Hashimoto’s thyroiditis: clinical results of a blinded placebo-controlled randomized prospective trial. J Endocrinol Invest 40:83–89

    CAS  Article  PubMed  Google Scholar 

  23. Bartalena L, Chiovato L, Vitti P (2016) Management of hyperthyroidism due to Graves’ disease: frequently asked questions and answers (if any). J Endocrinol Invest 39:1105–1114

    CAS  Article  PubMed  Google Scholar 

  24. Rotondo Dottore G, Leo M, Casini G, Latrofa F, Cestari L, Sellari-Franceschini S, Nardi M, Vitti P, Marcocci C, Marinò M (2017) Antioxidant actions of selenium in orbital fibroblasts: a basis for the effects of selenium in Graves’ orbitopathy. Thyroid 27:271–278

    CAS  Article  PubMed  Google Scholar 

  25. Tsai CC, Wu SB, Kao SC, Kau HC, Lee FL, Wei YH (2013) The protective effect of antioxidants on orbital fibroblasts from patients with Graves’ ophthalmopathy in response to oxidative stress. Mol Vis 19:927–934

    CAS  PubMed  PubMed Central  Google Scholar 

  26. Campo GM, Avenoso A, Campo S, Angela D, Ferlazzo AM, Calatroni A (2006) TNF-alpha, IFN-gamma, and IL-1beta modulate hyaluronan synthase expression in human skin fibroblasts: synergistic effect by concomital treatment with FeSO4 plus ascorbate. Mol Cell Biochem 292:169–178

    CAS  Article  PubMed  Google Scholar 

  27. Garlanda C, Dinarello CA, Mantovani A (2013) The interleukin-1 family: back to the future. Immunity 39:1003–1018

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  28. Bahn RS (2015) Current insights into the pathogenesis of Graves’ ophthalmopathy. Horm Metab Res 47:773–778

    CAS  Article  PubMed  Google Scholar 

  29. Rotondo Dottore G, Chiarini R, De Gregorio M, Leo M, Casini G, Cestari L, Sellari-Franceschini S, Nardi M, Vitti P, Marcocci C, Marinò M (2017) Selenium rescues orbital fibroblasts from cell death induced by hydrogen peroxide: another molecular basis for the effects of selenium in Graves’ orbitopathy. Endocrine (in press)

  30. Lisi S, Botta R, Lemmi M, Sellari-Franceschini S, Altea MA, Sisti E, Casini G, Nardi M, Marcocci C, Pinchera A, Marinò M (2011) Quercetin decreases proliferation of orbital fibroblasts and their release of hyaluronic acid. J Endocrinol Invest 34:521–527

    CAS  PubMed  Google Scholar 

  31. Tang XL, Liu XJ, Sun WM, Zhao J, Zheng RL (2005) Oxidative stress in Graves’ disease patients and antioxidant protection against lymphocytes DNA damage in vitro. Pharmazie 60:696–700

    CAS  PubMed  Google Scholar 

  32. Mourits MP, Prummel MF, Wiersinga WM, Koornneef L (1997) Clinical activity score as a guide in the management of patients with Graves’ ophthalmopathy. Clin Endocrinol (Oxf) 47:9–14

    CAS  Article  Google Scholar 

  33. Botta R, Lisi S, Marcocci C, Sellari-Franceschini S, Rocchi R, Latrofa F, Menconi F, Altea MA, Leo M, Sisti E, Casini G, Nardi M, Pinchera A, Vitti P, Marinò M (2013) Enalapril reduces proliferation and hyaluronic acid release in orbital fibroblasts. Thyroid 23:92–96

    CAS  Article  PubMed  Google Scholar 

  34. Levine M (1986) New concepts in the biology and biochemistry of ascorbic acid. N Engl J Med 314:892–902

    CAS  Article  PubMed  Google Scholar 

  35. Zafarullah M, Li WQ, Sylvester J, Ahmad M (2003) Molecular mechanisms of N-acetyl-l-cysteine actions. Cell Mol Life Sci 60:6–20

    CAS  Article  PubMed  Google Scholar 

  36. Hardeland R, Pandi-Perumal SR, Cardinali DP (2006) Melatonin. Int J Biochem Cell Biol 38:313–316

    CAS  Article  PubMed  Google Scholar 

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Affiliations

  1. Endocrinology Unit I, Department of Clinical and Experimental Medicine, University of Pisa and University Hospital of Pisa, Via Paradisa 2, 56124, Pisa, Italy

    G. Rotondo Dottore, I. Ionni, F. Menconi, P. Vitti, C. Marcocci & M. Marinò

  2. Ophthalmopathy Unit I, Department of Surgical, Medical and Molecular Pathology, University of Pisa and University Hospital of Pisa, Via Paradisa 2, 56124, Pisa, Italy

    G. Casini & M. Nardi

  3. ENT Unit I, Department of Surgical, Medical and Molecular Pathology, University of Pisa and University Hospital of Pisa, Via Paradisa 2, 56124, Pisa, Italy

    S. Sellari-Franceschini

Authors
  1. G. Rotondo Dottore
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  2. I. Ionni
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  3. F. Menconi
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  4. G. Casini
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  5. S. Sellari-Franceschini
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  6. M. Nardi
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  7. P. Vitti
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  8. C. Marcocci
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  9. M. Marinò
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Corresponding author

Correspondence to M. Marinò.

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All procedures performed in studies involving human participants were in accordance with the ethical standards of the institution and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

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Informed consent was obtained from all individual participants included in the study.

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Cite this article

Rotondo Dottore, G., Ionni, I., Menconi, F. et al. Action of three bioavailable antioxidants in orbital fibroblasts from patients with Graves’ orbitopathy (GO): a new frontier for GO treatment?. J Endocrinol Invest 41, 193–201 (2018). https://doi.org/10.1007/s40618-017-0718-7

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  • DOI: https://doi.org/10.1007/s40618-017-0718-7

Keywords

  • Antioxidant
  • Vitamin C
  • N-acetyl-l-cysteine
  • Melatonin
  • Orbitopathy
  • Ophthalmopathy
  • Graves’
  • Fibroblast