Korean J Physiol Pharmacol.  2020 Mar;24(2):149-156. 10.4196/kjpp.2020.24.2.149.

Inhibition of melanogenesis by sodium 2-mercaptoethanesulfonate

• Kim JH
• Oh CT
• Kwon TR
• Kim JH
• Bak DH
• Kim H
• Park WS
• Kim BJ

• KMID: 2471036
• DOI: http://doi.org/10.4196/kjpp.2020.24.2.149

Abstract

Sodium 2-mercaptoethanesulfonate (mesna) is a protective agent that is widely used in medicine because of its antioxidant effects. Recently, reactive oxygen species (ROS) were shown to increase pigmentation. Thus, ROS scavengers and inhibitors of ROS production may suppress melanogenesis. Forkhead box-O3a (FoxO3a) is an antimelanogenic factor that mediates ROS-induced skin pigmentation. In this study, we aimed to investigate the whitening effect of mesna and the signaling mechanism mediating this effect. Human melanoma (MNT-1) cells were used in this study. mRNA and protein expression were measured by real-time quantitative PCR and Western blotting analysis to track changes in FoxO3a-related signals induced by mesna. An immunofluorescence assay was performed to determine the nuclear translocation of FoxO3a. When MNT-1 melanoma cells were treated with mesna, melanin production and secretion decreased. These effects were accompanied by increases in FoxO3a activation and nuclear translocation, resulting in downregulation of four master genes of melanogenesis: MITF, TYR, TRP1, and TRP2. We found that mesna, an antioxidant and radical scavenger, suppresses melanin production and may therefore be a useful agent for the clinical treatment of hyperpigmentation disorders.

Keyword

Antioxidants; Hyperpigmentation; Mesna; Skin lightening preparations

MeSH Terms

Antioxidants
Blotting, Western
Down-Regulation
Fluorescent Antibody Technique
Humans
Hyperpigmentation
Melanins
Melanoma
Mesna*
Negotiating
Pigmentation
Polymerase Chain Reaction
Reactive Oxygen Species
RNA, Messenger
Skin Lightening Preparations
Skin Pigmentation
Sodium*
Antioxidants
Melanins
Mesna
RNA, Messenger
Reactive Oxygen Species
Skin Lightening Preparations
Sodium

Figure

• 

  Fig. 1 Effect of 2-mercaptoethanesulfonate (mesna) on 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging activity and mushroom tyrosinase activity. (A) The activity of DPPH scavenging of different concentrations of mesna and ascorbic acid (AA). The activities of mushroom tyrosinase assayed on (B) L-3,4-dihydroxyphenylalanine (L-DOPA) oxidation and (C) L-tyrosine hydroxylation in the presence of different concentrations of mesna and AA are shown. Results are expressed as means ± standard deviation of three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001 compared to non-treated.

• 

  Fig. 2 Effects of 2-mercaptoethanesulfonate (mesna) on melanogenesis in MNT-1 cells. (A) MNT-1 cells were treated with the mensa, N-acetylcysteine (NAC), and ascorbic acid (AA) for 72 h, and the melanin levels were visualized. (B) The quantitative melanin levels were determined by measuring absorbance at 490 nm using a spectrophotometer. (C) MNT-1 cell growth in the presence and absence of various mesna concentrations were compared. (D) MNT-1 cells were treated with mesna, NAC, and AA for 0–48 h. After treatment with the indicated reagents, gene expression was examined by quantitative real-time reverse-transcriptase-PCR. Results were normalized to those of GAPDH mRNA; measurements were calculated from triplicate experiments and are expressed ± standard deviation. *p < 0.05, **p < 0.01, ***p < 0.001 compared to non-treated cells. (E) MNT-1 cells were treated with mesna, NAC, and AA for 24–48 h. Whole cell lysates were analyzed by Western blotting using antibodies against MITF, TYR, TPR-1, and TRP-2. Equal sample loading was confirmed by probing with anti-β-actin antibodies. Results are expressed as averages of experiments performed in triplicate. *p < 0.05 compared to control.

• 

  Fig. 3 Effects of 2-mercaptoethanesulfonate (mesna) on the nuclear translocation of FoxO3a. (A) MNT-1 cells were treated with the indicated reagents (1 µM) for 24 h. The cells were then stained with anti-FoxO3a antibodies (green) and 4′,6-diamidino-2-phenylindole (DAPI) to stain nuclei (blue). Representative images were acquired using a fluorescence microscope and merged using DP Controller software (×200 magnification). (B) Cells with FoxO3a localization were counted. (C) MNT-1 cells were treated with mesna, N-acetylcysteine (NAC), and ascorbic acid (AA) for 24 h, after which cellular fractionation was performed and FoxO3a was analyzed by Western blotting. Results are expressed as averages of experiments performed in triplicate. **p < 0.01 compared to control.

• 

  Fig. 4 Effects of 2-mercaptoethanesulfonate (mesna) on MST1 and JNK phosphorylation. MNT-1 cells were treated with mesna, N-acetylcysteine (NAC), and ascorbic acid (AA) (1 µM) for the indicated time periods. Whole cell lysates were harvested and analyzed by Western blotting using antibodies against p-MST1, MST1, p-JNK, and JNK. Equal sample loading was confirmed by probing with antibodies against MST1, JNK, or β-actin. Results are expressed as averages of experiments performed in triplicate. *p < 0.05, **p < 0.01, ***p < 0.001 compared to 0 h.

Reference

1. Masaki H. Role of antioxidants in the skin: anti-aging effects. J Dermatol Sci. 2010; 58:85–90.

2. Meister A. Glutathione-ascorbic acid antioxidant system in animals. J Biol Chem. 1994; 269:9397–9400.

3. Villarama CD, Maibach HI. Glutathione as a depigmenting agent: an overview. Int J Cosmet Sci. 2005; 27:147–153.

4. Kabasakal L, Sehirli AO, Cetinel S, Cikler E, Gedik N, Sener G. Mesna (2-mercaptoethane sulfonate) prevents ischemia/reperfusion induced renal oxidative damage in rats. Life Sci. 2004; 75:2329–2340.

5. Yilmaz ER, Kertmen H, Gürer B, Kanat MA, Arikok AT, Ergüder BI, Hasturk AE, Ergil J, Sekerci Z. The protective effect of 2-mercaptoethane sulfonate (MESNA) against traumatic brain injury in rats. Acta Neurochir (Wien). 2013; 155:141–149.

6. Sakuma K, Ogawa M, Sugibayashi K, Yamada K, Yamamoto K. Relationship between tyrosinase inhibitory action and oxidationreduction potential of cosmetic whitening ingredients and phenol derivatives. Arch Pharm Res. 1999; 22:335–339.

7. Kim J, Choi H, Cho EG, Lee TR. FoxO3a is an antimelanogenic factor that mediates antioxidant-induced depigmentation. J Invest Dermatol. 2014; 134:1378–1388.

8. Wang H, Gao XD, Zhou GC, Cai L, Yao WB. In vitro and in vivo antioxidant activity of aqueous extract from Choerospondias axillaris fruit. Food Chem. 2008; 106:888–895.

9. Chang TS, Ding HY, Tai SS, Wu CY. Mushroom tyrosinase inhibitory effects of isoflavones isolated from soygerm koji fermented with Aspergillus oryzae BCRC 32288. Food Chem. 2007; 105:1430–1438.

10. Masamoto Y, Ando H, Murata Y, Shimoishi Y, Tada M, Takahata K. Mushroom tyrosinase inhibitory activity of esculetin isolated from seeds of Euphorbia lathyris L. Biosci Biotechnol Biochem. 2003; 67:631–634.

11. Hosoi J, Abe E, Suda T, Kuroki T. Regulation of melanin synthesis of B16 mouse melanoma cells by 1 alpha, 25-dihydroxyvitamin D3 and retinoic acid. Cancer Res. 1985; 45:1474–1478.

12. Yu T, Ji J, Guo YL. MST1 activation by curcumin mediates JNK activation, Foxo3a nuclear translocation and apoptosis in melanoma cells. Biochem Biophys Res Commun. 2013; 441:53–58.

13. Ura S, Masuyama N, Graves JD, Gotoh Y. MST1-JNK promotes apoptosis via caspase-dependent and independent pathways. Genes Cells. 2001; 6:519–530.

14. Skinner R, Sharkey IM, Pearson AD, Craft AW. Ifosfamide, mesna, and nephrotoxicity in children. J Clin Oncol. 1993; 11:173–190.

15. Choi J, Oh S, Lee D, Oh HJ, Park JY, Lee SB, Lim DS. Mst1-FoxO signaling protects Naïve T lymphocytes from cellular oxidative stress in mice. PLoS One. 2009; 4:e8011.

16. Sheth VM, Pandya AG. Melasma: a comprehensive update: part I. J Am Acad Dermatol. 2011; 65:689–697.

17. Rivas S, Pandya AG. Treatment of melasma with topical agents, peels and lasers: an evidence-based review. Am J Clin Dermatol. 2013; 14:359–376.

18. Ros JR, Rodríguez-López JN, García-Cánovas F. Effect of L-ascorbic acid on the monophenolase activity of tyrosinase. Biochem J. 1993; 295 (Pt 1):309–312.

19. Huh CH, Seo KI, Park JY, Lim JG, Eun HC, Park KC. A randomized, double-blind, placebo-controlled trial of vitamin C iontophoresis in melasma. Dermatology. 2003; 206:316–320.