Ann Dermatol.  2017 Feb;29(1):102-105. 10.5021/ad.2017.29.1.102.

Attenuation of Dickkopf 1-Induced Hair Growth Inhibition in Cultured Human Hair Follicles by Tianeptine

• Jeong HS
• Kwack MH
• Kim MK
• Kim JC
• Sung YK

• KMID: 2368038
• DOI: http://doi.org/10.5021/ad.2017.29.1.102

Abstract

No abstract available.

MeSH Terms

Hair Follicle*
Hair*
Humans*

Figure

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  Fig. 1 Effects of tianeptine on dickkopf 1 (DKK-1)-treated cultured hair follicles. (A) Isolated human hair follicles were cultured in the absence or presence of recombinant human DKK-1 (rhDKK-1) for 6 days and hair shaft elongation was measured. Values are means±standard deviation (SD) of seven determinations (from 7 hair follicles) per experiment from three independent experiments (*p<0.005). (B) Isolated human hair follicles were also cultured for 6 days in the absence or presence of 100 nM or 200 nM tianeptine together with rhDKK-1. Values are means±SD of ten determinations (from 10 hair follicles) per experiment from three independent experiments (*p<0.005). (C) Human hair follicles were cultured in the absence or presence of 100 nM tianeptine with 50 ng/ml rhDKK-1 for 3 days and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) labeling was performed (upper panel). Corresponding 4,6-diamidino-2-phenylindole (DAPI) nuclear staining is also shown (lower panel). (D) TUNEL positive apoptotic cells (green) in the hair bulb were counted and data are mean±SD from seven hair follicles (*p<0.005).

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  Fig. 2 Effects of tianeptine on cultured human dermal papilla (DP) cells and outer root sheath (ORS) cells. (A) DP cells were cultured in the presence or absence of tianeptine for 3 days and 3-[4,5] dimethylthiazol-2,5-diphenyltetrazolium bromide (MTT) assay was employed to measure the cell viability. Data are expressed as means±standard deviation (SD) of five determinations per experiment from four independent experiments using four different DP cell lines (*p<0.05 and **p<0.005). (B) DP cells were treated with tianeptine (10 and 100 nM) for 24 h and analyzed by real-time polymerase chain reaction (RT-PCR). Relative levels of genes are shown as mean±SD from three independent experiments (*p<0.05 and **p<0.005). (C) ORS cells were cultured in the presence or absence of tianeptine for 3 days and MTT data is shown. Data are expressed as means±SD of five determinations per experiment from three independent experiments using three different ORS cell lines. (D) ORS cells were treated with tianeptine (10 and 100 nM) for 24 h and analyzed by RT-PCR. For the amplification of insulin-like growth factor-1 (IGF-1), the forward primer 5′-TCAACAAGCCACAGGGTAT-3′ and reverse primer 5′-CGTGCAGAGCAAAGGAT-3′ were used. For the amplification of hepatocyte growth factor (HGF), the forward primer 5′-CGAGGCCATGGTGCTATACT-3′ and reverse primer 5′-ACACCAGGGTGATTCAGACC-3′ were used. For the amplification of vascular endothelial growth factor (VEGF), the forward primer 5′-TCTTCAAGCCATCCTGTGTG-3′ and reverse primer 5′-GCGAGTCTGTGTTTTTGCAG-3′ were used. For the amplification of keratinocyte growth factor (KGF), the forward primer 5′-GACATGGATCCTGCCAACTT-3′ and reverse primer 5′-AATTCCAACTGCCACTGTCC-3′ were used. For the amplification of β-actin, the forward primer 5′-GGACTTCGAGCAAGAGATGG-3′ and reverse primer 5′-AGCACTGTGTTGGCGTACAG-3′ were used. Amplification was performed under the following cycling conditions: 30 cycles (95℃ for 1 min, 60℃ for 1 min and 72℃ for 1 min).

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