Methylsulfonylmethane inhibits cortisol‑induced stress through p53‑mediated SDHA/HPRT1 expression in racehorse skeletal muscle cells: A primary step against exercise stress

Sp,Nipin; Kang,Dong  Young; Kim,Do  Hoon; Lee,Hyo  Gun; Park,Yeong‑Min; Kim,Il  Ho; Lee,Hak  Kyo; Cho,Byung‑Wook; Jang,Kyoung‑Jin; Yang,Young  Mok

doi:10.3892/etm.2019.8196

Methylsulfonylmethane inhibits cortisol‑induced stress through p53‑mediated SDHA/HPRT1 expression in racehorse skeletal muscle cells: A primary step against exercise stress

Authors:
- Nipin Sp
- Dong Young Kang
- Do Hoon Kim
- Hyo Gun Lee
- Yeong‑Min Park
- Il Ho Kim
- Hak Kyo Lee
- Byung‑Wook Cho
- Kyoung‑Jin Jang
- Young Mok Yang
View Affiliations

Affiliations: Department of Pathology, School of Medicine, Institute of Biomedical Science and Technology, Konkuk University, Chungju, Chungcheongbuk 27478, Republic of Korea, Department of Animal Science, College of Natural Resources and Life Sciences, Pusan National University, Miryang, Gyeongsangnam 50463, Republic of Korea, Department of Immunology, School of Medicine, Konkuk University, Chungju, Chungcheongbuk 27478, Republic of Korea, Nara Biotech Co., Ltd., Jeonju, Jeollabuk 54852, Republic of Korea, Department of Animal Biotechnology, Chonbuk National University, Jeonju, Jeollabuk 54896, Republic of Korea
Published online on: November 13, 2019 https://doi.org/10.3892/etm.2019.8196
Pages: 214-222
Copyright: © Sp et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Cortisol is a hormone involved in stress during exercise. The application of natural compounds is a new potential approach for controlling cortisol‑induced stress. Tumour suppressor protein p53 is activated during cellular stress. Succinate dehydrogenase complex subunit A (SDHA) and hypoxanthine phosphoribosyl transferase 1 (HPRT1) are considered to be two of the most stable reference genes when measuring stress during exercise in horses. In the present study cells were considered to be in a ʻstressed stateʼ if the levels of these stable genes and the highly stress responsive gene p53 were altered. It was hypothesized that a natural organic sulphur‑containing compound, methylsulfonylmethane (MSM), could inhibit cortisol‑induced stress in racing horse skeletal muscle cells by regulating SDHA, HPRT1 and p53 expression. After assessing cell viability using MTT assays, 20 µg/ml cortisol and 50 mM MSM were applied to horse skeletal muscle cell cultures. Reverse transcription‑quantitative PCR and western blot analysis demonstrated increases in SDHA, HPRT1 and p53 expression in cells in response to cortisol treatment, which was inhibited or normalized by MSM treatment. To determine the relationship between p53 and SDHA/HPRT1 expression at a transcriptional level, horse gene sequences of SDHA and HPRT1 were probed to identify novel binding sites for p53 in the gene promoters, which were confirmed using a chromatin immunoprecipitation assay. The relationship between p53 and SDHA/HPRT1 expression was confirmed using western blot analysis following the application of pifithrin‑α, a p53 inhibitor. These results suggested that MSM is a potential candidate drug for the inhibition of cortisol‑induced stress in racehorse skeletal muscle cells.

View Figures

View References

1	Williams CA: The effect of oxidative stress during exercise in the horse. J Anim Sci. 94:4067–4075. 2016. View Article : Google Scholar : PubMed/NCBI
2	Geor RJ and McCutcheon LJ: Hydration effects on physiological strain of horses during exercise-heat stress. J Appl Physiol (1985). 84:2042–2051. 1998. View Article : Google Scholar : PubMed/NCBI
3	Caillaud C, Connes P, Bouix D and Mercier J: Does haemorheology explain the paradox of hypoxemia during exercise in elite athletes or thoroughbred horses? Clin Hemorheol Microcirc. 26:175–181. 2002.PubMed/NCBI
4	González O, González E, Sánchez C, Pinto J, González I, Enríquez O, Martínez R, Filgueira G and White A: Effect of exercise on erythrocyte beta-adrenergic receptors and plasma concentrations of catecholamines and thyroid hormones in thoroughbred horses. Equine Vet J. 30:72–78. 1998. View Article : Google Scholar : PubMed/NCBI
5	Tadros EM, Frank N, De Witte FG and Boston RC: Effects of intravenous lipopolysaccharide infusion on glucose and insulin dynamics in horses with equine metabolic syndrome. Am J Vet Res. 74:1020–1029. 2013. View Article : Google Scholar : PubMed/NCBI
6	Morris MC and Rao U: Cortisol response to psychosocial stress during a depressive episode and remission. Stress. 17:51–58. 2014. View Article : Google Scholar : PubMed/NCBI
7	Álvarez-Diduk R and Galano A: Adrenaline and noradrenaline: Protectors against oxidative stress or molecular targets? J Phys Chem B. 119:3479–3491. 2015. View Article : Google Scholar : PubMed/NCBI
8	Flint MS, Baum A, Episcopo B, Knickelbein KZ, Liegey Dougall AJ, Chambers WH and Jenkins FJ: Chronic exposure to stress hormones promotes transformation and tumorigenicity of 3T3 mouse fibroblasts. Stress. 16:114–121. 2013. View Article : Google Scholar : PubMed/NCBI
9	Ranabir S and Reetu K: Stress and hormones. Indian J Endocrinol Metab. 15:18–22. 2011. View Article : Google Scholar : PubMed/NCBI
10	Konturek PC, Brzozowski T and Konturek SJ: Stress and the gut: Pathophysiology, clinical consequences, diagnostic approach and treatment options. J Physiol Pharmacol. 62:591–599. 2011.PubMed/NCBI
11	Mosavat M, Ooi FK and Mohamed M: Stress hormone and reproductive system in response to honey supplementation combined with different jumping exercise intensities in female rats. Biomed Res Int. 2014:1236402014. View Article : Google Scholar : PubMed/NCBI
12	Steimer T: The biology of fear- and anxiety-related behaviors. Dialogues Clin Neurosci. 4:231–249. 2002.PubMed/NCBI
13	McEwen BS: Central effects of stress hormones in health and disease: Understanding the protective and damaging effects of stress and stress mediators. Eur J Pharmacol. 583:174–185. 2008. View Article : Google Scholar : PubMed/NCBI
14	Hannibal KE and Bishop MD: Chronic stress, cortisol dysfunction, and pain: A psychoneuroendocrine rationale for stress management in pain rehabilitation. Phys Ther. 94:1816–1825. 2014. View Article : Google Scholar : PubMed/NCBI
15	Fioranelli M, Bottaccioli AG, Bottaccioli F, Bianchi M, Rovesti M and Roccia MG: Stress and inflammation in coronary artery disease: A review Psychoneuroendocrineimmunology-Based. Front Immunol. 9:20312018. View Article : Google Scholar : PubMed/NCBI
16	Butawan M, Benjamin RL and Bloomer RJ: Methylsulfonylmethane: Applications and safety of a novel dietary supplement. Nutrients. 9(pii): E2902017. View Article : Google Scholar : PubMed/NCBI
17	S P N, Darvin P, Yoo YB, Joung YH, Kang DY, Kim DN, Hwang TS, Kim SY, Kim WS, Lee HK, et al: The combination of methylsulfonylmethane and tamoxifen inhibits the Jak2/STAT5b pathway and synergistically inhibits tumor growth and metastasis in ER-positive breast cancer xenografts. BMC Cancer. 15:4742015. View Article : Google Scholar : PubMed/NCBI
18	Lim EJ, Hong DY, Park JH, Joung YH, Darvin P, Kim SY, Na YM, Hwang TS, Ye SK, Moon ES, et al: Methylsulfonylmethane suppresses breast cancer growth by down-regulating STAT3 and STAT5b pathways. PLoS One. 7:e333612012. View Article : Google Scholar : PubMed/NCBI
19	Joung YH, Darvin P, Kang DY, Sp N, Byun HJ, Lee CH, Lee HK and Yang YM: Methylsulfonylmethane inhibits RANKL-induced osteoclastogenesis in BMMs by suppressing NF-κB and STAT3 activities. PLoS One. 11:e01598912016. View Article : Google Scholar : PubMed/NCBI
20	Preetha NS, Kang DY, Darvin P, Kim DN, Joung YH, Kim SY, Cho KY, Do CH, Park KD, Lee JH, et al: Induction of in vitro ketosis condition and suppression using methylsulfonylmethane by altering ANGPTL3 expression through STAT5b signaling mechanism. Anim Cells Syst. 19:30–38. 2015. View Article : Google Scholar
21	Cappelli K, Felicetti M, Capomaccio S, Spinsanti G, Silvestrelli M and Supplizi AV: Exercise induced stress in horses: Selection of the most stable reference genes for quantitative RT-PCR normalization. BMC Mol Biol. 9:492008. View Article : Google Scholar : PubMed/NCBI
22	Cheng VW, Piragasam RS, Rothery RA, Maklashina E, Cecchini G and Weiner JH: Redox state of flavin adenine dinucleotide drives substrate binding and product release in Escherichia coli succinate dehydrogenase. Biochemistry. 54:1043–1052. 2015. View Article : Google Scholar : PubMed/NCBI
23	Guzy RD, Sharma B, Bell E, Chandel NS and Schumacker PT: Loss of the SdhB, but Not the SdhA, subunit of complex II triggers reactive oxygen species-dependent hypoxia-inducible factor activation and tumorigenesis. Mol Cell Biol. 28:718–731. 2008. View Article : Google Scholar : PubMed/NCBI
24	Gravells P, Ahrabi S, Vangala RK, Tomita K, Brash JT, Brustle LA, Chung C, Hong JM, Kaloudi A, Humphrey TC and Porter AC: Use of the HPRT gene to study nuclease-induced DNA double-strand break repair. Hum Mol Genet. 24:7097–7110. 2015.PubMed/NCBI
25	Ceballos-Picot I, Mockel L, Potier MC, Dauphinot L, Shirley TL, Torero-Ibad R, Fuchs J and Jinnah HA: Hypoxanthine-guanine phosphoribosyl transferase regulates early developmental programming of dopamine neurons: Implications for Lesch-Nyhan disease pathogenesis. Hum Mol Genet. 18:2317–2327. 2009. View Article : Google Scholar : PubMed/NCBI
26	Kang TH, Park Y, Bader JS and Friedmann T: The housekeeping gene hypoxanthine guanine phosphoribosyltransferase (HPRT) regulates multiple developmental and metabolic pathways of murine embryonic stem cell neuronal differentiation. PLoS One. 8:e749672013. View Article : Google Scholar : PubMed/NCBI
27	Surget S, Khoury MP and Bourdon JC: Uncovering the role of p53 splice variants in human malignancy: A clinical perspective. Onco Targets Ther. 7:57–68. 2013.PubMed/NCBI
28	Pflaum J, Schlosser S and Müller M: p53 family and cellular stress responses in cancer. Front Oncol. 4:2852014. View Article : Google Scholar : PubMed/NCBI
29	Han ES, Muller FL, Perez VI, Qi W, Liang H, Xi L, Fu C, Doyle E, Hickey M, Cornell J, et al: The in vivo gene expression signature of oxidative stress. Physiol Genomics. 34:112–126. 2008. View Article : Google Scholar : PubMed/NCBI
30	Alston CL, van der Westhuizen FH, He L, Wassmer E, Davison JE, Falkous G, McFarland R and Taylor RW: P53 Novel SDHA and SDHB mutations as a cause of isolated mitochondrial complex II deficiency. Neuromuscular Disord. 22 (Suppl 1):S212012. View Article : Google Scholar
31	Suzuki T, Kusunoki Y, Tsuyama N, Ohnishi H, Seyama T and Kyoizumi S: Elevated in vivo frequencies of mutant T cells with altered functional expression of the T-cell receptor or hypoxanthine phosphoribosyltransferase genes in p53-deficient mice. Mutat Res. 483:13–17. 2001. View Article : Google Scholar : PubMed/NCBI
32	Zhang H, Chi Y, Gao K, Zhang X and Yao J: p53 protein-mediated up-regulation of MAP kinase phosphatase 3 (MKP-3) contributes to the establishment of the cellular senescent phenotype through dephosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2). J Biol Chem. 290:1129–1140. 2015. View Article : Google Scholar : PubMed/NCBI
33	Maghsoudlou P, Ditchfield D, Klepacka DH, Shangaris P, Urbani L, Loukogeorgakis SP, Eaton S and De Coppi P: Isolation of esophageal stem cells with potential for therapy. Pediatr Surg Int. 30:1249–1256. 2014. View Article : Google Scholar : PubMed/NCBI
34	Joung YH, Lim EJ, Darvin P, Chung SC, Jang JW, Do Park K, Lee HK, Kim HS, Park T and Yang YM: MSM enhances GH signaling via the Jak2/STAT5b pathway in osteoblast-like cells and osteoblast differentiation through the activation of STAT5b in MSCs. PLoS One. 7:e474772012. View Article : Google Scholar : PubMed/NCBI
35	Kim DN, Joung YH, Darvin P, Kang DY, Sp N, Byun HJ, Cho KH, Park KD, Lee HK and Yang YM: Methylsulfonylmethane enhances BMP2induced osteoblast differentiation in mesenchymal stem cells. Mol Med Rep. 14:460–466. 2016. View Article : Google Scholar : PubMed/NCBI
36	Birch-Machin MA and Bowman A: Oxidative stress and ageing. Br J Dermatol. 175 (Suppl 2):S26–S29. 2016. View Article : Google Scholar
37	Walker DM, Patrick O'Neill J, Tyson FL and Walker VE: The stress response resolution assay. I. Quantitative assessment of environmental agent/condition effects on cellular stress resolution outcomes in epithelium. Environ Mol Mutagen. 54:268–280. 2013. View Article : Google Scholar : PubMed/NCBI
38	Walker DM, Nicklas JA and Walker VE: The stress response resolution assay. II. Quantitative assessment of environmental agent/condition effects on cellular stress resolution outcomes in epithelium. Environ Mol Mutagen. 54:281–293. 2013. View Article : Google Scholar : PubMed/NCBI
39	Morgan RA, Keen JA, Homer N, Nixon M, McKinnon-Garvin AM, Moses-Williams JA, Davis SR, Hadoke PWF and Walker BR: Dysregulation of cortisol metabolism in equine pituitary pars intermedia dysfunction. Endocrinology. 159:3791–3800. 2018. View Article : Google Scholar : PubMed/NCBI
40	Devi GR, Alam MJ and Singh RK: Synchronization in stress p53 network. Math Med Biol. 32:437–456. 2015.PubMed/NCBI
41	Karabay AZ, Koc A, Ozkan T, Hekmatshoar Y, Sunguroglu A, Aktan F and Buyukbingol Z: Methylsulfonylmethane induces p53 independent apoptosis in HCT-116 colon cancer cells. Int J Mol Sci. 17(pii): E11232016. View Article : Google Scholar : PubMed/NCBI
42	James A, Wang Y, Raje H, Rosby R and DiMario P: Nucleolar stress with and without p53. Nucleus. 5:402–426. 2014. View Article : Google Scholar : PubMed/NCBI