Korean J Physiol Pharmacol.  2017 May;21(3):287-292. 10.4196/kjpp.2017.21.3.287.

Pimecrolimus increases the melanogenesis and migration of melanocytes in vitro

Affiliations
  • 1First Clinical College, Nanjing University of Chinese Medicine, Nanjing 210029, China. tancheng@medmail.com.cn
  • 2Department of Dermatology, Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing 210029, China.
  • 3Department of Pathology, Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing 210029, China.

Abstract

Vitiligo is an intriguing depigmentary disorder and is notoriously difficult to be treated. The ultimate goal of vitiligo treatment is to replenish the lost melanocytes by immigration from hair follicle and to restore the normal function of melanogenesis by residual melanocytes. There are two types of topical calcineurin inhibitors called tacrolimus and pimecrolimus, and are recommended as the first-line treatments in vitiligo. Although pimecrolimus is efficacious for the repigmentation of vitiligo, its intrinsic mechanisms have never been investigated in vitro. This research aimed to study the ability of pimecrolimus on stimulating melanogenesis, melanocyte migration and MITF (microphthalmia associated transcription factor) protein expression. Results showed that pimecrolimus at the dosages of 1, 10, 10² nM were neither mitogenic nor cytotoxic to melanocytes. The addition of pimecrolimus at 10, 10² and 10³ nM significantly increased intracellular tyrosinase activity, which was consistent with the elevated content of melanin content at the same concentrations. The peak effect was seen at 72 h in response to 10² nM pimecrolimus. Results of the wound scratch assay and Transwell assays indicate that pimecrolimus is effective in facilitating melanocyte migration on a collagen IV-coated surface. In addition, MITF protein yield reached the highest by pimecrolimus at 10² nM. In brief, pimecrolimus enhances melanin synthesis as well as promotes migration of melanocytes directly, possibly via their effects on MITF protein expression.

Keyword

Calcineurin; Melanogenesis; Migration; Pimecrolimus; Vitiligo

MeSH Terms

Calcineurin
Calcineurin Inhibitors
Collagen
Emigration and Immigration
Hair Follicle
In Vitro Techniques*
Melanins
Melanocytes*
Microphthalmia-Associated Transcription Factor
Monophenol Monooxygenase
Tacrolimus
Vitiligo
Wounds and Injuries
Calcineurin
Calcineurin Inhibitors
Collagen
Melanins
Microphthalmia-Associated Transcription Factor
Monophenol Monooxygenase
Tacrolimus

Figure

  • Fig. 1 Effects of pimecrolimus on the proliferation (A), tyrosinase activity (B) and melanin content (C) of human epidermal melanocytes.Cells were cultured for three days with pimecrolimus at various concentrations (1, 10, 100, and 1,000 nM) and compared with the control. Results represent the mean±SD of three independent experiments. *p<0.05 compared with the control.

  • Fig. 2 Effects of pimecrolimus on melanocytic migration in vitro.(A) Migration was measured by scratch assay. The average gap was evaluated three days after incubation with pimecrolimus or the control. Results represent the mean±SD of three independent experiments. (B) Migration was measured by Transwell assay. Cells pretreated with pimecrolimus and the control were stained with crystal violet after they penetrated to the lower surface of the membrane. The immigrated cells were counted under a light microscope at ×200. Results represent the mean±SD of three independent experiments. *p<0.05 compared with the control.

  • Fig. 3 Effect of pimecrolimus on the protein expression levels of MITF in melanocytes.The protein expression level of MITF was examined by Western blotting after a 3-day incubation with 10 or 100 nM pimecrolimus. Results represent the mean±SD of three independent experiments. *p<0.05 compared with the control.


Reference

1. Whitton ME, Pinart M, Batchelor J, Leonardi-Bee J, González U, Jiyad Z, Eleftheriadou V, Ezzedine K. Interventions for vitiligo. Cochrane Database Syst Rev. 2015; (2):CD003263. PMID: 25710794.
Article
2. Chou WC, Takeo M, Rabbani P, Hu H, Lee W, Chung YR, Carucci J, Overbeek P, Ito M. Direct migration of follicular melanocyte stem cells to the epidermis after wounding or UVB irradiation is dependent on Mc1r signaling. Nat Med. 2013; 19:924–929. PMID: 23749232.
Article
3. Guerra L, Dellambra E, Brescia S, Raskovic D. Vitiligo: pathogenetic hypotheses and targets for current therapies. Curr Drug Metab. 2010; 11:451–467. PMID: 20540698.
Article
4. Taieb A, Alomar A, Böhm M, Dell'anna ML, De Pase A, Eleftheriadou V, Ezzedine K, Gauthier Y, Gawkrodger DJ, Jouary T, Leone G, Moretti S, Nieuweboer-Krobotova L, Olsson MJ, Parsad D, Passeron T, Tanew A, van der Veen W, van Geel N, Whitton M, Wolkerstorfer A, Picardo M. Vitiligo European Task Force (VETF). European Academy of Dermatology and Venereology (EADV). Union Europe´enne des Me´decins Spe´cialistes (UEMS). Guidelines for the management of vitiligo: the European Dermatology Forum consensus. Br J Dermatol. 2013; 168:5–19. PMID: 22860621.
Article
5. Khalid M, Mujtaba G, Haroon TS. Comparison of 0.05% clobetasol propionate cream and topical Puvasol in childhood vitiligo. Int J Dermatol. 1995; 34:203–205. PMID: 7751099.
Article
6. Ezzedine K, Eleftheriadou V, Whitton M, van Geel N. Vitiligo. Lancet. 2015; 386:74–84. PMID: 25596811.
Article
7. Kathuria S, Khaitan BK, Ramam M, Sharma VK. Segmental vitiligo: a randomized controlled trial to evaluate efficacy and safety of 0.1% tacrolimus ointment vs 0.05% fluticasone propionate cream. Indian J Dermatol Venereol Leprol. 2012; 78:68–73. PMID: 22199063.
Article
8. Köse O, Arca E, Kurumlu Z. Mometasone cream versus pimecrolimus cream for the treatment of childhood localized vitiligo. J Dermatolog Treat. 2010; 21:133–139. PMID: 20394489.
Article
9. Leone G, Pacifico A, Iacovelli P, Paro Vidolin A, Picardo M. Tacalcitol and narrow-band phototherapy in patients with vitiligo. Clin Exp Dermatol. 2006; 31:200–205. PMID: 16487090.
Article
10. Kostovic K, Pasic A. New treatment modalities for vitiligo: focus on topical immunomodulators. Drugs. 2005; 65:447–459. PMID: 15733009.
11. Travis LB, Weinberg JM, Silverberg NB. Successful treatment of vitiligo with 0.1% tacrolimus ointment. Arch Dermatol. 2003; 139:571–574. discussion 573. PMID: 12756092.
Article
12. Dawid M, Veensalu M, Grassberger M, Wolff K. Efficacy and safety of pimecrolimus cream 1% in adult patients with vitiligo: results of a randomized, double-blind, vehicle-controlled study. J Dtsch Dermatol Ges. 2006; 4:942–946. PMID: 17081269.
Article
13. Bilaç DB, Ermertcan AT, Sahin MT, Oztürkcan S. Two therapeutic challenges: facial vitiligo successfully treated with 1% pimecrolimus cream and 0.005% calcipotriol cream. J Eur Acad Dermatol Venereol. 2009; 23:72–73. PMID: 18355205.
Article
14. Shim WH, Suh SW, Jwa SW, Song M, Kim HS, Ko HC, Kim BS, Kim MB. A pilot study of 1% pimecrolimus cream for the treatment of childhood segmental vitiligo. Ann Dermatol. 2013; 25:168–172. PMID: 23717007.
Article
15. Lan CC, Chen GS, Chiou MH, Wu CS, Chang CH, Yu HS. FK506 promotes melanocyte and melanoblast growth and creates a favourable milieu for cell migration via keratinocytes: possible mechanisms of how tacrolimus ointment induces repigmentation in patients with vitiligo. Br J Dermatol. 2005; 153:498–505. PMID: 16120133.
Article
16. Kang HY, Choi YM. FK506 increases pigmentation and migration of human melanocytes. Br J Dermatol. 2006; 155:1037–1040. PMID: 17034537.
Article
17. Tan C, Zhu W, Lu Y. Aloin, cinnamic acid and sophorcarpidine are potent inhibitors of tyrosinase. Chin Med J (Engl). 2002; 115:1859–1862. PMID: 12622939.
18. Lee BW, Schwartz RA, Hercogová J, Valle Y, Lotti TM. Vitiligo road map. Dermatol Ther. 2012; 25(Suppl 1):S44–S56. PMID: 23237038.
Article
19. Falabella R, Barona MI. Update on skin repigmentation therapies in vitiligo. Pigment Cell Melanoma Res. 2009; 22:42–65. PMID: 19040503.
Article
20. Fiorentino DF, Chen RO, Stewart DB, Brown KK, Sundram UN. The direct cellular target of topically applied pimecrolimus may not be infiltrating lymphocytes. Br J Dermatol. 2011; 164:996–1003. PMID: 21166661.
Article
21. Smit NP, Van Rossum HH, Romijn FP, Sellar KJ, Breetveld M, Gibbs S, Van Pelt J. Calcineurin activity and inhibition in skin and (epi) dermal cell cultures. J Invest Dermatol. 2008; 128:1686–1690. PMID: 18200052.
22. Lee JY, Kang WH. Effect of cyclosporin A on melanogenesis in cultured human melanocytes. Pigment Cell Res. 2003; 16:504–508. PMID: 12950728.
Article
23. Lee KY, Jeon SY, Hong JW, Choi KW, Lee CY, Choi SJ, Kim JH, Song KH, Kim KH. Endothelin-1 enhances the proliferation of normal human melanocytes in a paradoxical manner from the TNF-α-inhibited condition, but tacrolimus promotes exclusively the cellular migration without proliferation: a proposed action mechanism for combination therapy of phototherapy and topical tacrolimus in vitiligo treatment. J Eur Acad Dermatol Venereol. 2013; 27:609–616. PMID: 22404745.
24. Jung H, Chung H, Chang SE, Kang DH, Oh ES. FK506 regulates pigmentation by maturing the melanosome and facilitating their transfer to keratinocytes. Pigment Cell Melanoma Res. 2016; 29:199–209. PMID: 26581186.
Article
25. Coskun B, Saral Y, Turgut D. Topical 0.05% clobetasol propionate versus 1% pimecrolimus ointment in vitiligo. Eur J Dermatol. 2005; 15:88–91. PMID: 15757818.
26. Wong R, Lin AN. Efficacy of topical calcineurin inhibitors in vitiligo. Int J Dermatol. 2013; 52:491–496. PMID: 23331250.
Article
27. Rao C, Su Z, Li H, Ma X, Zheng X, Liu Y, Lu F, Qu J, Hou L. Microphthalmia-associated transcription factor regulates skin melanoblast migration by repressing the melanoma cell adhesion molecule. Exp Dermatol. 2016; 25:74–76. PMID: 26284703.
Article
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