Ann Dermatol.  2015 Dec;27(6):667-675. 10.5021/ad.2015.27.6.667.

Herbal Extracts Induce Dermal Papilla Cell Proliferation of Human Hair Follicles

• Rastegar H
• Ashtiani H
• Aghaei M
• Barikbin B
• Ehsani A

• KMID: 2157441
• DOI: http://doi.org/10.5021/ad.2015.27.6.667

Abstract

BACKGROUND
The number of people suffering from balding or hair thinning is increasing, despite the advances in various medical therapies. Therefore, it is highly important to develop new therapies to inhibit balding and increase hair proliferation.
OBJECTIVE
We investigated the effects of herbal extracts commonly used for improving balding in traditional medicine to identify potential agents for hair proliferation.
METHODS
The expression levels of 5alpha-reductase isoforms (type I and II) were analyzed using quantitative real-time reverse transcription polymerase chain reaction in the human follicular dermal papilla cells (DPCs). The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenylteterazolium bromide and bromodeoxyuridine tests were used to evaluate the cell proliferation effect of herbal extracts in DPCs. The expression levels of extracellular signal-regulated kinase (ERK), Akt, cyclin D1, cyclin-dependent kinase 4 (Cdk4), B-cell lymphoma (Bcl-2) and Bcl-2-associated X protein (Bax) were measured using western blot analysis.
RESULTS
The 5alpha-reductase isoform mRNAs and proteins were detected in the cultured DPCs, and the expression level of 5alpha-R2 in DPCs in the presence of the herbal extracts was gradually decreased. Herbal extracts were found to significantly increase the proliferation of human DPCs at concentrations ranging from 1.5% to 4.5%. These results show that the herbal extracts tested affected the protein expressions of ERK, Akt, cyclin D1, Cdk4, Bcl-2, and Bax in DPCs.
CONCLUSION
These results suggest that herbal extracts exert positive effects on hair proliferation via ERK, Akt, cyclin D1, and Cdk4 signaling in DPCs; they also suggest that herbal extracts could be a great alternative therapy for increasing hair proliferation.

Keyword

Dermal papilla cell; Hair follicle; Herbal extracts; Proliferation

MeSH Terms

bcl-2-Associated X Protein
Blotting, Western
Bromodeoxyuridine
Cell Proliferation*
Cyclin D1
Cyclin-Dependent Kinase 4
Hair Follicle*
Hair*
Humans*
Lymphoma, B-Cell
Medicine, Traditional
Phosphotransferases
Polymerase Chain Reaction
Protein Isoforms
Reverse Transcription
RNA, Messenger
Bromodeoxyuridine
Cyclin D1
Cyclin-Dependent Kinase 4
Phosphotransferases
Protein Isoforms
RNA, Messenger
bcl-2-Associated X Protein

Figure

• 

  Fig. 1 Relative gene expression of two 5α-reductase (5αR) isoforms in cultured human dermal papilla cells (DPCs). (A) Relative gene expression of 5αR1 and 5αR2 were detected using quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR) in DPCs. Data represent three independent experiments and relative expression values were calculated using the equation RQ=2-ΔΔct. (B) qPCR analysis of 5αR1 and 5αR2 mRNA expression in DPCs following herbal extract treatment using graded concentrations. Expression levels were normalized to human glyceraldehyde 3-phosphate dehydrogenase (GAPDH) mRNA. Data are mean±standard deviation; *p<0.05 and †p<0.01 compared with untreated control group.

• 

  Fig. 2 Effects of herbal extracts on expression of 5α-reductase (5αR) isoform proteins in human dermal papilla cells (DPCs). Cells were treated with indicated concentrations of herbal extracts for 48 hours and then expression of proteins was analyzed using western blotting. SRD5A1: steroid 5αR type I, GAPDH: glyceraldehyde 3-phosphate dehydrogenase.

• 

  Fig. 3 Effect of herbal extracts on cell proliferation of human dermal papilla cells (DPCs). Cells were treated with different concentrations of herbal extracts with or without Akt kinase inhibitor for 48 hours, and proliferation was assessed using (A) 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and (B) bromodeoxyuridine (BrdU) assays. Herbal extracts increased cell proliferation in DPCs concentration-dependently. Results are mean±standard deviation and were calculated as a percentage of corresponding control values. *p<0.05 and †p<0.01, one-way analysis of variance. Each point represents four repetitions each in triplicate.

• 

  Fig. 4 Effects of herbal extracts on expression of cell cycle regulatory proteins in human dermal papilla cells (DPCs). Cells were treated with indicated concentrations of herbal extracts for 48 hours and then the expression of proteins was analyzed using western blotting. Cdk4: cyclin-dependent kinase 4, GAPDH: glyceraldehyde 3-phosphate dehydrogenase.

• 

  Fig. 5 Herbal extracts increased phosphorylated extracellular signal-regulated kinase (p-ERK) and p-Akt in cultured dermal papilla cells (DPCs) analyzed using western blot. Level of p-ERK and p-Akt increased after 48 hours treatment concentrationdependently. GAPDH: glyceraldehyde 3-phosphate dehydrogenase.

• 

  Fig. 6 Effects of herbal extracts on expression of apoptosis-related proteins in human dermal papilla cells (DPCs). Cells were treated with indicated concentrations of herbal extracts for 48 hours and then expression of proteins was analyzed using western blotting. Bcl-2: B-cell lymphoma, Bax: Bcl-2-associated X protein, GAPDH: glyceraldehyde 3-phosphate dehydrogenase.

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