Hair Growth Effect of DN106212 in C57BL/6 Mouse and Its Network Pharmacological Mechanism of Action

Abstract

Centipeda minima (CMX) has been widely investigated using network pharmacology and clinical studies for its effects on hair growth via the JAK/STAT signaling pathway. Human hair follicle papilla cells exhibit hair regrowth through the expression of Wnt signaling-related proteins. However, the mechanism of action of CMX in animals has not been elucidated fully. This study examined the effect of induced hair loss and its side-effects on the skin, and observed the mechanism of action of an alcoholic extract of CMX (DN106212) on C57BL/6 mice. Our results showed that DN106212 was more effective in promoting hair growth than dimethyl sulfoxide in the negative control and tofacitinib (TF) in the positive control when mice were treated with DN106212 for 16 days. We confirmed that DN106212 promotes the formation of mature hair follicles through hematoxylin and eosin staining. We also found that the expression of vascular endothelial growth factor (Vegfa), insulin-like growth factor 1 (Igf1), and transforming growth factor beta 1 (Tgfb1) is related to hair growth using PCR. DN106212-treated mice had significantly higher expression of Vegfa and Igf1 than TF-treated ones, and inhibiting the expression of Tgfb1 had similar effects as TF treatment. In conclusion, we propose that DN106212 increases the expression of hair growth factors, promotes the development of hair follicles, and promotes hair growth. Although additional experiments are needed, DN106212 may serve as an experimental basis for research on natural hair growth-promoting agents.

Keywords: Centipeda minima; DN106212; Igf1; Vegfa; hair growth.

Figure 1

HPLC chromatograms of DN106212. Overlapped HPLC chromatogram of the compound DN106212 and the CMX fraction used in previous study (grey line) (Kim et al. 2021 [34]) and an enhanced purification (blue line).

Figure 2

Chemical structures of arnicolide D, arnicolide C, microhelenin C, and brevilin A.

Figure 3

The effect of DN106212 on hair growth in C57BL/6 mice. (a) Indicated doses of DN106212 and TF were applied topically to the dorsal skin twice a day for 16 days. (b) Comparison of percentage of area hair regrowth on days 0, 7, 10, 13, and 16 among all groups. Data are presented as the mean ± SD, n = 6 in each group; * p < 0.05 (compared with CTL), ^# p < 0.05 (compared with TF).

Figure 4

Results of 16 day application of topical control (a), TF (b), and DN106212 (c). Different hair growth patterns can be observed. The arrow indicates that the hair follicle is generated in the deep dermis. Longitudinal sections of the dorsal skin (16 day) were stained using H&E and viewed under 40× magnification. Scale bar: 100 µm. Each image presented is a representative of six mice.

Figure 5

Comparison of the expression of Vegfa, Igf1, and Tgfb1 mRNA evaluated with RT-PCR. The relative expression levels of (a) Vegfa, (b) Igf1, and (c) Tgfb1 were determined using qPCR analysis. * p < 0.05, ** p < 0.01, and *** p < 0.001 compared with the control group.

Figure 6

Compound–target network of CMX. Boxes and circles represent the compounds and targets, respectively. Edges between compounds and targets indicate the interactions between them. The targets of CMX were colored to indicate three pathways with a high combined score among pathways related to hair loss. CMX, emulsion extract from Centipeda minima.