Ann Dermatol.  2019 Apr;31(2):164-174. 10.5021/ad.2019.31.2.164.

Synthesized Ceramide Induces Growth of Dermal Papilla Cells with Potential Contribution to Hair Growth

• Oh JH
• Jeong KH
• Kim JE
• Kang H

• KMID: 2439062
• DOI: http://doi.org/10.5021/ad.2019.31.2.164

Abstract

BACKGROUND
The ceramide is known to play an important role in the formation of intracellular lipids, and play a crucial role as a barrier for skin and hair cuticle. Recent study has revealed that ceramide has potential effect on hair growth in a mouse model. However, the role of ceramide in human dermal papilla cells (hDPCs) known to play an important role in hair growth is not well understood yet.
OBJECTIVE
The goal of this study was to investigate the effect of synthetic ceramides (oleyl and stearyl ceramides) on hair growth using hDPCs.
METHODS
hDPCs were treated with synthesized ceramides. hDPCs viability was evaluated by MTT assay. The expression of hair growth related factors were investigated by western blot, real-time polymerase chain reaction and growth factor array. The expression of β-catenin was confirmed by immunofluorescence.
RESULTS
Treatment with ceramides increased the expression of proteins affecting cell proliferation such as Bcl-2, BAX, phosphorylated-ERK and Cyclin D1. Also, ceramides treatment were increased the expression of several growth factors, including epidermal growth factor family, and promote the expression of Wnt/β-catenin and BMP2/4 signaling.
CONCLUSION
Our data suggest that synthetic ceramides stimulates hair growth by induction proliferation of hDPCs via modulation of Wnt/β-catenin and BMP2/4 signaling.

Keyword

Ceramide; Dermal papilla cell; Hair; Wnt signaling pathway

MeSH Terms

Animals
Blotting, Western
Cell Proliferation
Ceramides
Cyclin D1
Epidermal Growth Factor
Fluorescent Antibody Technique
Hair*
Humans
Intercellular Signaling Peptides and Proteins
Mice
Real-Time Polymerase Chain Reaction
Skin
Wnt Signaling Pathway
Ceramides
Cyclin D1
Epidermal Growth Factor
Intercellular Signaling Peptides and Proteins

Figure

• 

  Fig. 1 Effects of ceramides on viability of human dermal papilla cells (hDPCs). (A) Scheme of ceramide's synthesis process. (B) Chemical structures of synthetic ceramides (oleyl and stearyl ceramide). (C) The effect of ceramides on hDPCs viability was determined by MTT assay. hDPCs were cultured for 24 hours. After starvation, these cultured hDPCs were treated with three kinds of ceramides for 72 hours. Stearyl Cer: stearyl ceramide, Oleyl Cer: oleyl ceramide. *p<0.05 compared with C8-ceramide treatment group, **p<0.01 compared with group treated with C8-ceramide (positive control).

• 

  Fig. 2 Ceramide increases expression levels of Bcl-2/BAX and ERK/Cyclin-D1. (A) Expression levels of various factors known to affect proliferation were examined by western blot. Bcl-2 expression in groups treated with oleyl and stearyl ceramides was increased compare to those of the control group and C8-ceramide group. (B~D) Phosphorylated-ERK was normalized relative to total ERK. Cyclin-D1 was normalized relative to β-actin. Western blot results showed upregulated levels of Cyclin-D1 and phosphorylated ERK after treatment with ceramides compare to the control group. Results are expressed as mean±standard deviation. C8-Cer: C8-ceramide, Stearyl Cer: stearyl ceramide, Oleyl Cer: oleyl ceramide. *p<0.05 compared with the control, **p<0.01 compared with the control, ***p<0.001 compared with the control.

• 

  Fig. 3 (A) Ceramides upregulate growth factor protein levels. Human growth factor antibody array was used to detect growth factor expression in human dermal papilla cells (hDPCs). (B) Ceramide treatment significantly increased the expression of growth factors in hDPCs. Results are expressed as mean±standard deviation. C8-Cer: C8-ceramide, Stearyl Cer: stearyl ceramide, Oleyl Cer: oleyl ceramide, AR: amphiregulin, IGF-1: insulin-like growth factor 1, EGF: epidermal growth factor, EGFR: receptor.. *p<0.05 compared with the control, **p<0.01 compared with the control.

• 

  Fig. 4 Effects of ceramide treatment on B-catenin pathway. Phosphorylation of Akt/GSK3β and β-catenin expression was determined by western blot. Graph was schematized data of western blot band. (A, B) Phosphorylation of Akt/GSK3β and β-catenin expression was increased by ceramide treatment compare to those in the control. (C) β-catenin expression in the nucleus of human dermal papilla cells was determined by immunofluorescence staining. Oleyl and stearyl ceramides significantly increased β-catenin expression in nuclei compare to C8-ceramide treatment or control. DAPI (blue) was used to counterstain nuclei (×100). Results are expressed as mean±standard deviation. C8-Cer: C8-ceramide, Stearyl Cer: stearyl ceramide, Oleyl Cer: oleyl ceramide. *p<0.05 compared with the control.

• 

  Fig. 5 Ceramides upregulate expression of Wnt/β-catenin targets but downregulate expression of BMP target. Expression levels of target genes were determined using real-time polymerase chain reaction. (A) Wnt target genes: Wnt3, Wnt5a, Wnt7a, Wnt10b; (B) β-catenin target genes: β-catenin, AP-1, Lef-1, Axin2; and (C) BMP target genes: BMP2, BMP4. Results are expressed as mean±standard deviation. C8-Cer: C8-ceramide, Stearyl Cer: stearyl ceramide, Oleyl Cer: oleyl ceramide. *p<0.05 compared with the control, **p<0.01, ***p<0.001 compared with the control, #p<0.05 compared with the group treated with C8-ceramide.

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