Ann Dermatol.  2018 Dec;30(6):694-700. 10.5021/ad.2018.30.6.694.

The Effects of the 3-OH Group of Kaempferol on Interfollicular Epidermal Stem Cell Fate

• Chae JB
• Choi HR
• Shin JW
• Na JI
• Huh CH
• Park KC

• KMID: 2428925
• DOI: http://doi.org/10.5021/ad.2018.30.6.694

Abstract

BACKGROUND
Kaempferol (3,4"²,5,7-tetrahydroxyflavone) is a flavonoid known to have a wide range of pharmacological activities. The 3-OH group in flavonoids has been reported to determine antioxidant activities.
OBJECTIVE
We tested whether kaempferol can affect the expression of integrins and the stem cell fate of interfollicular epidermal stem cells.
METHODS
Skin equivalent (SE) models were constructed, and the expression levels of stem cell markers and basement membrane-related antigens were tested. The immunohistochemical staining patterns of integrins, p63, and proliferating cell nuclear antigen (PCNA) were compared between kaempferol- and apigenin-treated SE models. Reverse transcription-polymerase chain reaction (RT-PCR) was used to evaluate the mRNA expression of integrins.
RESULTS
Kaempferol increased the thickness of the epidermis when added to prepare SEs. In addition, the basal cells of kaempferol- treated SEs appeared more columnar. In the immunohistological study, the expression of integrins α6 and β1 and the numbers of p63- and PCNA-positive cells were markedly higher in the kaempferol-treated model. However, apigenin showed no effects on the formation of three-dimensional skin models. RT-PCR analysis also confirmed that kaempferol increased the expression of integrin α6 and integrin β1.
CONCLUSION
Our findings indicated that kaempferol can increase the proliferative potential of basal epidermal cells by modulating the basement membrane. In other words, kaempferol can affect the fate of interfollicular epidermal stem cells by increasing the expression of both integrins α6 and β1. These effects, in particular, might be ascribed to the 3-OH group of kaempferol.

Keyword

Basement membrane; Extracellular matrix; Kaempferol; Stem cell; 3-OH

MeSH Terms

Apigenin
Basement Membrane
Epidermis
Extracellular Matrix
Flavonoids
Integrins
Proliferating Cell Nuclear Antigen
RNA, Messenger
Skin
Stem Cells*
Apigenin
Flavonoids
Integrins
Proliferating Cell Nuclear Antigen
RNA, Messenger

Figure

• 

  Fig. 1 The molecular structures of kaempferol and apigenin. Their structures are the same except for the presence and absence, respectively, of the 3-OH functional group (A, B). Kaempferol demonstrated satisfactory antioxidant activity from a concentration of 0.025 mM onwards in the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay, while apigenin did not (C).

• 

  Fig. 2 Cytotoxicity of kaempferol (A, B) and apigenin (C, D) to cultured normal keratinocytes and fibroblasts. Cells were treated as described in the “materials and methods.” The y-axis represents the relative cell viability following the MTT assay. The values shown are the means±standard deviation of triplicate wells.

• 

  Fig. 3 The effects of kaempferol and apigenin on skin equivalent reconstruction and integrin expression. Sections of skin equivalents were stained for hematoxylin and eosin (H&E) (A~C), integrin α6 (E~G), and integrin β1 (I~K). Stained sections were analyzed with an image analysis program (D, H, L). Experiments were performed twice with the same cells; representative results are shown (original magnification: ×400, **p<0.01 in comparison with the control group).

• 

  Fig. 4 Immunohistochemical staining for proliferative markers and involucrin. Sections were stained for p63 (A~C), proliferating cell nuclear antigen (PCNA) (E~G), and involucrin (I~K), respectively. The expression levels were analyzed with an image analysis program (D, H, L). Experiments were performed twice with the same cells; representative results are shown (original magnification: ×400, *p<0.05; **p<0.01 in comparison with the control group).

• 

  Fig. 5 Reverse transcription-polymerase chain reaction analysis for ITGA6 and ITGB1 expression levels. The expression of both integrins was increased by kaempferol treatment in a dose-dependent manner (**p<0.01 in comparison with the control group).

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