L–Thyroxine induced hyperthyroidism effect on Testicular endoplasmic reticulum stress and Perk–mediated Nrf2/HO–1 signaling axis of rats

• Gözde Arkalı Firat University, Faculty of Veterinary Medicine, Department of Physiology. Elazığ, Türkiye
• Şeyma Özer Kaya Firat University, Faculty of Veterinary Medicine, Department of Reproduction and Artificial Insemination. Elazığ, Türkiye
• Songül Çeribaşı Firat University, Faculty of Veterinary Medicine, Department of Pathology. Elazığ, Türkiye
• Edanur Güler Ekmen Firat University, Faculty of Veterinary Medicine, Department of Physiology. Elazığ, Türkiye
• Mesut Aksakal Firat University, Faculty of Veterinary Medicine, Department of Physiology. Elazığ, Türkiye
• Mehmet Çay Firat University, Faculty of Veterinary Medicine, Department of Physiology. Elazığ, Türkiye

DOI: https://doi.org/10.52973/rcfcv-e34436

Keywords: Hyperthyroidism, endoplasmic reticulum stress, Nrf2, HO–1, infertility

Abstract

The effect of thyroid gland on the male reproductive system (neonatal–prepubertal and adult periods) has been investigated for many years. Hypertyhroidism may cause male infertility by effecting spermatological parameters such as loss of motility and decreased of sperm concentration. However, the mechanisms of male infertility caused by hyperthyroidism are still not fully explained. The aim of this study was to investigate the effect of hyperthyroidism on testicular endoplasmic reticulum stress and the protein kinase RNA–like endoplasmic reticulum kinase (PERK) mediated antioxidant pathway in adult male rats. 24 Sprague Dawley adult male rats were used. Rats were divided into two groups: control group (received intraperitoneal injections of saline solution 1 mL·day^-1 for 8 week) and hyperthyroidism group (received intraperitoneal injections of l–thyroxine 0,3 mg·kg^-1·mL^-1·day^-1 for 8 week). The serum free triiodothyronine (fT3) (P<0.01) and free thyroxine (fT4) (P<0.05) levels were increased, thyroid stimulating hormone (TSH) level (P<0.01) and final body weight (P<0.001) were decreased in the hyperthyroid groups. It was determined that tubulus seminiferus contortus diameters, germinal cell thicknesses, johnsen testicular score values significantly decreased in the hyperthyroid group (P<0.001). It was determined that it had a negative effect on reproductive organs weight and spermatological parameters. As per our results, hyperthyroidism significantly increased malondialdehyde level (P<0.01), glutathione level (P<0.001), glutathione peroxidase enzyme activity (P<0.001), PERK, GRP78 (P<0.01), ATF4 (P<0.05), Nrf2, HO–1 (P<0.05) protein expression levels and significantly decreased catalase activity (P<0.05). These results showed that increased thyroid hormones levels may be a negative factor in terms of testicular physiology as it causes endoplasmic reticulum stress in the testes, and PERK mediated antioxidant response may play an important role in testicular tissue in hyperthyroidism.

References

Shahid MA, Ashraf MA, Sharma S. Physiology, Thyroid Hormone [Internet]. Treasure Island (FL, USA): StatPearls Publishing; 2024 [cited 18 Jun. 2023]; 8 p. PMID 29763182. Available in: https://goo.su/UDZljk

Doubleday AR, Sippel RS. Hyperthyroidism. Gland. Surg. [Internet]. 2020; 9(1):124–135. doi: https://doi.org/grxmbf

Holsberger DR, Cooke PS. Understanding the role of thyroid hormone in Sertoli cell development: a mechanistic hypothesis. Cell Tissue Res. [Internet]. 2005; 322:133–140. doi: https://doi.org/drxcrn

Mendis–Handagama SM, Siril–Ariyaratne HB. Leydig cells, thyroid hormones and steroidogenesis. Indian J. Exp. Biol. [Internet]. 2005 [cited 18 Feb. 2024]; 43(11):939–962. PMID: 16313060. Available in: https://goo.su/iPqMq4W

Ouriquea GM, Finamor IA, Saccol EMH, Riffel APK, Pês TS, Gutierrez K, Gonçalves PBD, Baldisserotto B, Pavanato MA, Barreto KP. Resveratrol improves sperm motility, prevents lipid peroxidation and enhances antioxidant defences in the testes of hyperthyroid rats. Reprod. Toxicol. [Internet]. 2013; 37:31–39. doi: https://doi.org/g8n6xt

Asker ME, Hassan WA, El–Kashlan AM. Experimentally induced hyperthyroidism influences oxidant and antioxidant status and impairs male gonadal functions in adult rats. Andrologia [Internet]. 2015; 47(6):644–654. doi: https://doi.org/f7h6k3

Wajner SM, Wagner MS, Maia AL. Clinical implicationsof altered thyroid status in male testicular function. Arq. Bras. Endocrinol. Metab. [Internet]. 2009; 53(8):976–982. doi: https://doi.org/fcdf27

Krajewska–Kulak E, Sengupta P. Thyroid function in male infertility. Front. Endocrinol. [Internet]. 2013; 4(174)1–2. doi: https://doi.org/g8n6xv

Maiorino M, Ursini F. Oxidative stress, spermatogenesis and fertility. Biol. Chem. [Internet]. 2002; 38(3–4):591–597. doi: https://doi.org/bx3hmb

Venditti P, Di Meo S. Thyroid hormone–induced oxidative stress. Cell Mol. Life Sci. [Internet]. 2006; 63(4):414–434. doi: https://doi.org/c8bzfq

Moazamian R, Polhemus A, Connaughton H, Fraser B, Whiting S, Gharagozloo P, Aitken RJ. Oxidative stress and human spermatozoa: Diagnostic and functional significance of aldehydes generated as a result of lipid peroxidation. Mol. Human. Rep. [Internet]. 2015; 21(6):502–515. doi: https://doi.org/f7fsws

Mogulkoc R, Baltaci AK, Oztekin E, Aydin L, Tuncer I. Hyperthyroidism causes lipid peroxidation in kidney and testis tissues of rats: Protective role of melatonin. Neuro Endocrinol. Lett. [Internet]. 2005 [cited 15 Jan. 2024]; 26(6):806–810. PMID: 16380687. Available in: https://goo.su/b9yAAL

Zamoner A, Barreto KP, Wilhelm Filho D, Sell F, Woehl VM, Guma FCR, Silva FRMB, Pessoa–Pureur R. Hyperthyroidism in the developing rat testis is associated with oxidative stress and hyperphosphorylated vimentin accumulation. Mol. Cell. Endocrinol. [Internet]. 2007; 267(1–2):116–126. doi: https://doi.org/dx2mrd

Schröder M, Kaufman RJ. The mammalian unfolded protein response. Annu. Rev. Biochem. [Internet]. 2005; 74:739–789. doi: https://doi.org/cdxq32

Karna KK, Shin YS, Choi BR, Kim HK, Park KJ. The role of endoplasmic reticulum stress response in male reproductive physiology and pathology: A review. World J. Mens. Health [Internet]. 2020; 38(4):484–494. doi: https://doi.org/gp6vnd

Clermont Y. Kinetics of spermatogenesis in mammals: seminiferous epithelium cycle and spermatogonial renewal. Physiol. Rev. [Internet]. 1972; 52(1):198–236. doi: https://doi.org/g8n6xw

Bancroft JD, Stevens A. Theory and practice of histological techniques. 3th ed. London: Churchill Livingstone; 1990. 744 p.

Johnsen SG. Testicular biopsy score count – a method for registration of spermatogenesis in human testes: normal values and results in 335 hypogonadal males. Hormones [Internet]. 1970; 1(1):2–25. doi: https://doi.org/bvxfrs

Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with Folin phenol reagent J. Biol. Chem. [Internet]. 1951; 193(1):265–275. PMID: 14907713. doi: https://doi.org/ghv6nr

Placer ZA, Cushman LL, Johnson BC. Estimation of product of lipid peroxidation (malonly dialdehyde) in biochemical systems. Anal. Biochem. 1966; 16(2):359–364. doi: https://doi.org/b96rpj

Sedlak J, Lindsay RH. Estimation of total protein bound and nonprotein sulfhydryl groups in tissue with Ellman’s reagent. Anal. Biochem. [Internet] 1968; 25:192–205. doi: https://doi.org/csbsfm

Lawrence RA, Burk RF. Glutathione peroxidase activity in selenium–deficient rat liver. Biochem. Biophys. Res. Commun. [Internet]. 1976; 71(4):952–958. doi: https://doi.org/d3vv59

Aebi H. Catalase in vitro. Methods Enzymol. [Internet]. 1984; 105:121–126. doi: https://doi.org/dnf7v9

Bass JJ, Wilkinson DJ, Rankin D, Phillips BE, Szewczyk NJ, Smith K, Atherton PJ. An overview of technical considerations for Western blotting applications to physiological research. Scand. J. Med. Sci. Sports [Internet]. 2017; 27(1):4–25. doi: https://doi.org/f9knh5

Türk G, Ateşşahin A, Sönmez M, Yüce A, Çeribaşı AO. Lycopene protects against cyclosporine A–induced testicular toxicity in rats. Theriogenology [Internet]. 2007; 67(4):778–785. doi: https://doi.org/fxghnh

Shahat AS, Hassan WA, El–Sayed WM. N–Acetylcysteine and Safranal prevented the brain damage induced by hyperthyroidism in adult male rats. Nutr. Neurosci. [Internet]. 2022; 25(2):231–245. doi: https://doi.org/g8n6xx

Ireton–Jones CS. Intake: Energy. In: Mahan LK, Escott–Stump S, Raymond JL, editors. Krause’s food & the nutrition care process. 13th ed. St Louis (MO, USA): Elsevier Saunders, 2012; 19–31 p.

Bartalena L, Bogazzi F, Brogioni S, Burelli A, Scarcello G, Martino E. Measurement of serum free thyroid hormone concentrations: an essential tool for the diagnosis of thyroid dysfunction. Horm. Res. [Internet]. 1996; 45(3–5):142–147. doi: https://doi.org/ff72dx

Asayama K, Dobashi K, Hayashibe H, Megata Y, Kato K. Lipid peroxidation and free radical scavengers in thyroid dysfunction in the rat: A possible mechanism of injury to heart and skeletal muscle in hyperthyroidism. Endocrinology [Internet]. 1987; 121(6):2112–2118. doi: https://doi.org/d5q4kf

Huh K, Kwon TH, Kim JS, Park JM. Role of the hepatic xanthine oxidase in thyroid dysfunction: effect of thyroid hormones in oxidative stress in rat liver. Arch. Pharm. Res. [Internet]. 1998; 21:236–249. doi: https://doi.org/b8bpd8

Civelek S, Seymen O, Seven A, Yigit G, Hatemi H, Burçak G. Oxidative stress in heart tissue of hyperthyroid and iron supplemented rats. J. Toxicol. Environ. Health A. 2001; 64(6):499–506. doi: https://doi.org/bvpxc7

Sahoo DK, Roy A, Bhanja S, Chainy GBN. Experimental hyperthyroidism–induced oxidative stress and impairment of antioxidant defence system in rat testis. Indian J. Exp. Biol. [Internet]. 2005 [cited 18 Jan. 2024]; 43(11):1058–1067. PMID: 16313068. Available in: https://goo.su/z3Z23V

Sahoo DK, Roy A, Chainy GBN. Protetive effect of vitamin E and curcumin on L–thyroxine–induced rat testicular oxidative stress. Chem. Biol. Interact. [Internet]. 2008; 176(2–3):121–128. doi: https://doi.org/b3gtz7

Chattopadhyay S, Sahoo DK, Subudhi U, Chainy GBN. Differential expression profiles of antioxidant enzymes and glutathione redox status in hyperthyroid rats: a temporal analysis. Comp. Biochem. Physiol. C Toxicol. Pharmacol. [Internet]. 2007; 146(3):383–391. doi: https://doi.org/crv374

Abo–Elnour RK, El–Deeb FD. A histological study on the effect of experimentally induced hyperthyroidism on adult albino rat testis. Egypt. J. Histol. [Internet]. 2012 [cited 25 Feb. 2024]; 35(4):862–871. Available en: https://goo.su/91Tnd

Özgüner M, Şenol A, Ural M, İşler M. Deneysel hipertiroidinin erişkin sıçan testis dokusunda oluşturduğu histolojik değişiklikler [Histologic changes of adult rat testis in experimental hyperthyroidism]. SDÜ. Tıp. Fak. Derg. [Internet]. 2009 [cited 18 Jan. 2024];119(4):1–6. Turkish. Available in: https://goo.su/UmUrx

Faraone–Mennella MR, Ferone A, Marino L, Cardone A, Comitato R, Venditti P, Di Meo S, Farina B. Poly(ADP–ribosyl)ation of proteins and germ cell developmentin hyperthyroid testis. Mol. Cell. Biochem. [Internet]. 2009; 323(1–2):119–129. doi: https://doi.org/bgm528

Khosrowbeygi A, Zarghami N, Deldar Y. Correlation between sperm quality parameters and seminal plasma antioxidants status. Iran J. Reprod. Med. [Internet]. 2004 [cited 20 Feb. 2024]; 2(2):58–64. Available in: https://goo.su/35DN0

Wagner MS, Wajner SM, Maia AL. The Role of thyroid hormone in testicular development and function. J. Endocrinol. [Internet]. 2008; 199(3): 351–365. doi: https://doi.org/fnjjf8

Kim JH, Park SJ, Kim TS, Park HJ, Park J, Kim BK, Kim GR, Kim JM, Huang SM, Chae J, Park CK, Lee DS. Testicular hyperthermia induces Unfolded Protein Response signaling activation in spermatocyte. Biochem Biophys. Res. Commun. [Internet]. 2013; 434(4):861–866. doi: https://doi.org/f4x3xv

Tabuchi Y, Takasaki I, Kondo T. Identification of genetic networks involved in the cell injury accompanying endoplasmic reticulum stress induced by bisphenol A in testicular Sertoli cells. Biochem. Biophys Res. Commun. [Internet]. 2006; 345(3):1044–1050. doi: https://doi.org/b2hqcg

Araujo AS, Fernandes T, Ribeiro MF, Khaper N, Belló–Klein A. Redox regulation of myocardial ERK 1/2 phosphorylation in experimental hyperthyroidism: role of Thioredoxin–Peroxiredoxin system. J. Cardiovasc. Pharmacol. [Internet]. 2010; 56(5):513–517. doi: https://doi.org/dnc34r

Zhao P, Hu Z, Ma W, Zang L, Tian Z, Hou Q. Quercetin alleviates hyperthyroidism–induced liver damage via Nrf2 signaling pathway. BioFactors [Internet]. 2020; 46(4):608–619. doi: https://doi.org/gm3wq3

Brunt KR, Tsuji MR, Lai JH, Kinobe RT, Durante W, Claycomb WC, Ward CA, Melo LG. Heme oxygenase–1 inhibits pro–oxidant induced hypertrophy in HL–1 cardiomyocytes. Exp. Biol. Med. [Internet]. 2009; 234(5):582–594. doi: https://doi.org/cpg8kr

Harding HP, Zhang Y, Zeng H, Novoa I, Lu PD, Calfon M, Sadri N, Yun C, Popko B, Paules R, Stojdl DF, Bell JC, Hettmann T, Leiden JM, Ron D. An integrated stress response regulates amino acid metabolism and resistance to oxidative stress. Mol. Cell [Internet]. 2003; 11(3):619–633. doi: https://doi.org/bqzdfn

Bektur NE, Sahin E, Kaçar S, Bağci R, Karakaya S, Dönmez DB, Şahintürk V. Investigation of the effect of hyperthyroidism on endoplasmic reticulum stress and transient receptor potential canonical 1 channel in the kidney. Turk J. Med. Sci. [Internet]. 2021; 51(3):1554–1563. doi: https://doi.org/g8n6x2

Aykanat NEB, Sahin E, Kaçar S, Bağci R, Karakaya S, Dönmez DB, Şahintürk V. Cardiac hypertrophy caused by hyperthyroidism in rats: the role of ATF–6 and TRPC1 channels. Can. J. Physiol. Pharmacol. [Internet]. 2021; 99(11):1226–1233. doi: https://doi.org/grgnrb

Liang B, Liu LY, Huang HB, Li LY, Zhou JX. High T3 induces beta–cell insulin resistance via endoplasmic reticulum stress. Mediators Inflamm. [Internet]. 2020;2020: 5287108. doi: https://doi.org/g8n6x3

Lin JH, Li H, Zhang YH, Ron D, Walter P. Divergent Effects of PERK and IRE1 signaling on cell viability. PLos One [Internet]. 2009; 4(1):e4170. doi: https://doi.org/bd47h2

Rutkowski DT, Arnold SM, Miller CN, Wu J, Li J, Gunnison KM, Mori K, Sadighi Akha AA, Raden D, Kaufman RJ. Adaptation to ER stress is mediated by differential stabilities of pro–survival and pro–apoptotic mRNAs and proteins. PLoS Biol. [Internet]. 2006;4(11):e374. doi: https://doi.org/c3rwkc