Ann Dermatol.  2016 Feb;28(1):55-64. 10.5021/ad.2016.28.1.55.

Role of Arachidonic Acid in Promoting Hair Growth

• Munkhbayar S
• Jang S
• Cho AR
• Choi SJ
• Shin CY
• Eun HC
• Kim KH
• Kwon O

• KMID: 2429482
• DOI: http://doi.org/10.5021/ad.2016.28.1.55

Abstract

BACKGROUND
Arachidonic acid (AA) is an omega-6 polyunsaturated fatty acid present in all mammalian cell membranes, and involved in the regulation of many cellular processes, including cell survival, angiogenesis, and mitogenesis. The dermal papilla, composed of specialized fibroblasts located in the bulb of the hair follicle, contributes to the control of hair growth and the hair cycle.
OBJECTIVE
This study investigated the effect of AA on hair growth by using in vivo and in vitro models.
METHODS
The effect of AA on human dermal papilla cells (hDPCs) and hair shaft elongation was evaluated by MTT assay and hair follicle organ culture, respectively. The expression of various growth and survival factors in hDPCs were investigated by western blot or immunohistochemistry. The ability of AA to induce and prolong anagen phase in C57BL/6 mice was analyzed.
RESULTS
AA was found to enhance the viability of hDPCs and promote the expression of several factors responsible for hair growth, including fibroblast growth factor-7 (FGF-7) and FGF-10. Western blotting identified the role of AA in the phosphorylation of various transcription factors (ERK, CREB, and AKT) and increased expression of Bcl-2 in hDPCs. In addition, AA significantly promoted hair shaft elongation, with increased proliferation of matrix keratinocytes, during ex vivo hair follicle culture. It was also found to promote hair growth by induction and prolongation of anagen phase in telogen-stage C57BL/6 mice.
CONCLUSION
This study concludes that AA plays a role in promoting hair growth by increasing the expression of growth factors in hDPCs and enhancing follicle proliferation and survival.

Keyword

Arachidonic acid; Hair; Hair follicle

MeSH Terms

Animals
Arachidonic Acid*
Blotting, Western
Cell Membrane
Cell Survival
Fibroblasts
Hair Follicle
Hair*
Humans
Immunohistochemistry
Intercellular Signaling Peptides and Proteins
Keratinocytes
Mice
Organ Culture Techniques
Phosphorylation
Transcription Factors
Arachidonic Acid
Intercellular Signaling Peptides and Proteins
Transcription Factors

Figure

• 

  Fig. 1 The effect of arachidonic acid (AA) on viability of cultured human dermal papilla cells (hDPCs). (A) Treatment of hDPCs with AA (1~5 µM) resulted in significantly increased cell viability, as measured by the MTT assay. (B) AA-enhanced hair shaft elongation in ex vivo follicle culture. Human hair follicles (HFs) were treated with the vehicle or AA (1, 2, 5, 10 µM) (n=5); AA significantly enhanced hair shaft elongation. (C, D) The proliferation of matrix keratinocytes increases in AA-treated HFs. Human HFs were cultured with the vehicle or AA (1, 2 µM, or 5 µM) for 3 days, and then subjected to immunofluorescence staining, to examine proliferation in the hair matrix keratinocytes, with Ki-67 (proliferation, green fluorescence), and 4',6-diamidino-2-phenylindole (DAPI, blue fluorescence) to counterstain the nuclei (×200). Data are expressed as mean±standard error. CTL: control. *p≤0.05, **p≤0.01, ***p≤0.01, versus the control group.

• 

  Fig. 2 Arachidonic acid (AA) increases the expression of growth and survival factor genes in human dermal papillary cells (hDPCs). (A) The mRNA for fibroblast growth factors (FGF-7 and -10) and hepatocyte growth factor (HGF) is elevated in hDPCs that were treated with AA (1, 2 µM, or 5 µM) for 24 hours. (B) Treatment with AA increases the expression of FGF-7 and FGF-10 in the cytoplasm of the dermal papilla (DP) and inner root sheath (IRS) on day 3 of its application, compared to the vehicle control (×100). (C and D) Human DPCs were lysed to analyze the total protein content of the lysates via Western blotting by using primary antibodies for total-p42/44 ERK, phospho-p42/44 ERK, total-AKT, phospho-AKT, total-CREB, phospho-CREB, Bcl-2, and β-actin. ERK: extracellular signal-regulated kinases, AKT: protein kinase B, CREB: cAMP response element-binding protein, CTL, control. Data are expressed as mean±standard error. *p≤0.05, **p≤0.01, ***p≤0.01, versus the control group.

• 

  Fig. 3 Enhanced anagen induction in response to arachidonic acid (AA) treatment in 8-week-old female C57BL/6 mice. (A) After their backs were shaved, five mice in each group, were treated either with the vehicle, topical 3% minoxidil (MNX) (positive control), or 2% AA for 4 weeks. (B) Weekly representation of the amount of black colored skin. (C) Weekly representation of the amount of hair-bearing skin. Data are expressed as mean±standard error. CTL: control, D0: first day after the back was shaved, W: week.*p≤0.05, ***p≤0.001, versus the control group.

• 

  Fig. 4 Arachidonic acid (AA) prolongs the anagen hair cycle. (A) The experiment design. Ten days after depilation by waxing, the back skin of C57BL/6 mice was treated with either with the vehicle, topical 3% minoxidil (MNX), or 2% AA. (B) On day 21 after depilation, biopsied skin samples were fixed in paraffin, sectioned and stained with hematoxylin and eosin (×40). (C) Calculating the hair cycle scores. For each mouse (5 mice per group), 50 hair follicles (HFs) from each section were analyzed and graded as follows: anagen VI, 100; early catagen, 200; mid catagen, 300; late catagen, 400. The score indicates the mean hair cycle stage for all HFs in a group. Data are expressed as mean±standard error. CTL: control. *p≤0.05, ***p≤0.001 versus the control group.

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