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

• Xu P
• Chen J
• Tan C
• Lai RS
• Min ZS

• KMID: 2376956
• DOI: http://doi.org/10.4196/kjpp.2017.21.3.287

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.

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