Methylsulfonylmethane inhibits cortisol-induced stress through p53-mediated SDHA/HPRT1 expression in racehorse skeletal muscle cells: A primary step against exercise stress.
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 () and hypoxanthine phosphoribosyl transferase 1 () 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 'stressed state' 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 and 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 and / expression at a transcriptional level, horse gene sequences of and 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.
Copyright: © Sp et al.
Publication Date: 2019-11-13 PubMed ID: 31853292PubMed Central: PMC6909739DOI: 10.3892/etm.2019.8196Google Scholar: Lookup
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Summary
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This research article examines how a natural organic sulphur-containing compound, methylsulfonylmethane (MSM), can inhibit stress induced by the hormone cortisol in racehorse skeletal muscle cells, focusing on the effects on two stable stress-measuring genes and a highly responsive stress gene.
Mechanism of Stress Measurement
- The researchers considered cells to be in a stressed state if the levels of two specific genes—Succinate dehydrogenase complex subunit A (SDHA) and hypoxanthine phosphoribosyl transferase 1 (HPRT1)—and the highly stress-responsive gene p53 were altered. This is because SDHA and HPRT1 are known to be relatively stable under typical conditions, so fluctuations in their levels could suggest a response to stress.
The Role of Cortisol and MSM
- The researchers initiated stress in horse skeletal muscle cells by adding 20 µg/ml of cortisol to their cultures. They hypothesized that MSM, at a dose of 50 mM, could normalize this cortisol-induced stress.
- They used MTT assays to measure cell viability after treatment with cortisol and MSM, and found that MSM was able to inhibit the cortisol-induced stress.
Analyzing Expression of SDHA, HPRT1, and p53
- Expression levels of SDHA, HPRT1, and p53 in cells were examined through reverse transcription-quantitative PCR and western blot analysis. Results demonstrated increases in the expression of these genes in response to cortisol treatment.
- This cortisol-triggered increase was then normalized or inhibited by adding MSM to the cells, thus verifying their initial hypothesis.
Confirmation of the p53 and SDHA/HPRT1 Relationship
- To establish a correlation between p53 and SDHA/HPRT1, the researchers studied the horse gene sequences of SDHA and HPRT1 for potential new binding sites for p53 in their respective gene promoters. The presence of such sites was confirmed using a chromatin immunoprecipitation assay.
- The connection between p53 and the expression of SDHA and HPRT1 was further substantiated using western blot analysis following the use of pifithrin-α, a molecule known for inhibiting p53.
Conclusions
- These findings suggest that MSM could be a viable option for quelling cortisol-induced stress in racehorse skeletal muscle cells. Hence, MSM might help to counteract exercise-induced stress in racehorses, strengthening endurance during high-stress racing events.
Cite This Article
APA
Sp N, Kang DY, Kim DH, Lee HG, Park YM, Kim IH, Lee HK, Cho BW, Jang KJ, Yang YM.
(2019).
Methylsulfonylmethane inhibits cortisol-induced stress through p53-mediated SDHA/HPRT1 expression in racehorse skeletal muscle cells: A primary step against exercise stress.
Exp Ther Med, 19(1), 214-222.
https://doi.org/10.3892/etm.2019.8196 Publication
Researcher Affiliations
- Department of Pathology, School of Medicine, Institute of Biomedical Science and Technology, Konkuk University, Chungju, Chungcheongbuk 27478, Republic of Korea.
- Department of Pathology, School of Medicine, Institute of Biomedical Science and Technology, Konkuk University, Chungju, Chungcheongbuk 27478, Republic of Korea.
- 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.
- Department of Animal Science, College of Natural Resources and Life Sciences, Pusan National University, Miryang, Gyeongsangnam 50463, Republic of Korea.
- Department of Pathology, School of Medicine, Institute of Biomedical Science and Technology, Konkuk University, Chungju, Chungcheongbuk 27478, Republic of Korea.
- Department of Pathology, School of Medicine, Institute of Biomedical Science and Technology, Konkuk University, Chungju, Chungcheongbuk 27478, Republic of Korea.
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