Ann Dermatol.  2010 Aug;22(3):255-261. 10.5021/ad.2010.22.3.255.

Enhancement of Keratinocyte Differentiation by Rose Absolute Oil

• Kim JH
• Choi DK
• Lee SS
• Choi SJ
• Kim CD
• Yoon TJ
• Lee JH

• KMID: 2265357
• DOI: http://doi.org/10.5021/ad.2010.22.3.255

Abstract

BACKGROUND
Through differentiation processes, keratinocytes provide a physical barrier to our bodies and control skin features such as moisturization, wrinkles and pigmentation. Keratinocyte differentiation is disturbed in several skin diseases such as psoriasis and atopic dermatitis.
OBJECTIVE
The aim of this study is to evaluate the keratinocyte differentiation-enhancing effect of rose absolute oil (RAO).
METHODS
Primary cultured human normal keratinocytes were treated with RAO, and differentiation then checked by the expression of marker genes.
RESULTS
RAO did not induce cytotoxicity on cultured keratinocytes at a dose of 10microM. The level of involucrin, an early marker for keratinocyte differentiation, was significantly increased by RAO. Concomitantly, RAO increased involucrin promoter activity, indicating that RAO increased involucrin gene expression at the mRNA level. Furthermore, RAO increased the level of filaggrin in cultured keratinocytes, and in the granular layer of mouse skin. In line with these results, RAO decreased the proliferation of keratinocytes cultured in vitro. When RAO was applied topically on the tape-stripped mouse skins, it accelerated the recovery of disturbed barrier function.
CONCLUSION
These results suggest that RAO may be applicable for the control of skin texture and keratinocyte differentiation-related skin diseases.

Keyword

Differentiation; Filaggrin; Keratinocyte; Rose absolute oil

MeSH Terms

Animals
Dermatitis, Atopic
Gene Expression
Humans
Intermediate Filament Proteins
Keratinocytes
Mice
Pigmentation
Protein Precursors
Psoriasis
RNA, Messenger
Skin
Skin Diseases
Intermediate Filament Proteins
Protein Precursors
RNA, Messenger

Figure

• 

  Fig. 1 Cytotoxicity of rose absolute oil (RAO). Normal human epidermal keratinocytes were treated with RAO at the indicated concentrations for 3 days. Cell viability was measured by MTT assay. The mean values±SEM are averages of triplicate measurements.

• 

  Fig. 2 Effect of rose absolute oil (RAO) on involucrin expression in keratinocytes. (A) Normal human epidermal keratinocytes were treated with RAO at the indicated concentrations for 3 days. Calcium (1.2 mM) was included as a positive control for keratinocyte differentiation. Cellular proteins were prepared and protein level for involucrin was evaluated by Western blot. Actin was used as a loading control. (B) Effect of RAO on the involucrin promoter activity. Keratinocytes were transduced with 10 multiplicity of infections (MOIs) of involucrin-luc reporter adenovirus, then treated with RAO for 3 days. Cells were lysed and assayed for luciferase activity. Data are represented as relative light unit (RLU) and SEM, measured from 3 independent experiments.

• 

  Fig. 3 Effect of rose absolute oil (RAO) on filaggrin expression. Normal human epidermal keratinocytes were treated with RAO at the indicated concentrations for 7 days. Calcium (1.2 mM) was included as a positive control for keratinocyte differentiation. Cellular proteins were prepared and protein level for filaggrin was evaluated by Western blot. Actin was used as a loading control.

• 

  Fig. 4 Effect of rose absolute oil (RAO) on the cell growth. Normal human epidermal keratinocytes were treated with 10µg/ml RAO for the indicated time points in the presence of 1µCi of [3H]thymidine. Calcium (1.2 mM) was included as a positive control for keratinocyte differentiation. Cells were lysed using 0.1 N NaOH and radioactivity was measured. Data are expressed as percentage of control. The mean values±SEM are averages of triplicate measurements.

• 

  Fig. 5 Effect of rose absolute oil (RAO) on transepidermal water loss (TEWL). (A) Hairless mice were tape-stripped, then topically treated with 0.1% solution of RAO (in 70% polyethylene glycol and 30% ethanol) and RAO complex in which RAO (0.1%), oleic acid (0.1%), oleanolic acid (0.05%), urocanic acid (0.01%), acetylhexapeptide (0.01%) and ceramide (0.1%) were additionally included. At the indicated time points, TEWL was measured using Tewameter. Basal TEWL (0 h) was measured before tape-stripping. Five hairless mice were included in each group. Data are represented as average and SEM. (B) Skin specimens were obtained after 5 days of treatment. Sections were prepared and stained with anti-filaggrin antibody (×100).

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