JKM > Volume 39(1); 2018 > Article
Kwon, Kim, and Yang: Effects of Topical application of Astragalus membranaceus in Spontaneous Alopecia Mice Model

Abstract

Objectives

Astragalus membranaceus has been reported to inhibit immune responses, but its effect on hair loss is not clear. In this study, the effect of A. membranaceus extract (AM) on hair regrowth in C57BL/6 mice with natural hair loss in the telogen phase was investigated.

Methods

Mice with natural hair loss were topically treated with 1% AM on the dorsal skin for 2 weeks. Dorsal skin samples were stained with hematoxylin and eosin and probed with an anti-mouse CD8a IgG. The mRNA expression levels of tumor necrosis factor (TNF)-α, interferon (IFN)-γ and interleukin (IL)-4 were measured by reverse transcription polymerase chain reaction and quantitative real-time polymerase chain reaction.

Results

AM treatment induced hair regrowth in hair loss mice, while control mice suffered continued hair loss. Tapering hair shafts and broken hair follicles were decreased as well as CD8+ T lymphocyte infiltration. In addition, the expressions of TNF-α, IFN-γ and IL-4 were reduced by AM treatment. Also, AM treatment significantly increased the KGF expressions in Hs68 fibroblast cells.

Conclusion

These results suggest that topical application of A. membranaceus may be an alternative therapy for hair loss.

Fig. 1
Quality evaluation of Astragalus membranaceus. HPLC chromatograms of standard formononetin (A) and A. membranaceus extract (B). It takes 31.7 min for the retention times of formononetin itself and formononetin in A. membranaceus extract.
jkm-39-1-1f1.gif
Fig. 2
A. membranaceus induces hair regrowth in hair loss mice. Morphological fingdings on the back of the C57BL/6J mice. The mice of A. membranaceus group were treated with 100 μ L of 1% (w/v) A. membranaceus for 14 days. Following challenge for 15 days, photographs were taken and mice were sacrificed. Normal, non-treated normal control mice; Control, vehicle-treated alopecia mice; AM, AM-treated alopecia mice.
jkm-39-1-1f2.gif
Fig. 3
A. membranaceus recovers the dystrophy of hair follicles. Histological findings by hematoxylin and eosin (H&E) staining of dorsal skin sections (n = 7). The magnifications were × 100 (upper panel) and × 200 (lower panel). Arrow indicates ‘swarm of bees’. Arrowhead indicates tapering hair shaft. Normal, non-treated normal control mice; Control, vehicle-treated alopecia mice; AM, AM-treated alopecia mice.
jkm-39-1-1f3.gif
Fig. 4
A. membranaceus reduces the infiltration of CD8+ cells in skin tissues. Immunohistochemical CD8+ cell staining. Arrow indicates brown-stained CD8+ cells. Normal, non-treated normal control mice; Control, vehicle-treated alopecia mice; AM, AM-treated alopecia mice.
jkm-39-1-1f4.gif
Fig. 5
A. membranaceus decreases the mRNA expressions of cytokines in skin tissues. Expression of TNF-α, IL-4 and IFN-γ mRNA levels by RT-PCR (A) and qPCR (B). Results are presented as mean ± S.E.M. ## and ### indicates the mean differs significantly between Normal group and Control group (p < 0.01 and p < 0.001, respectively). *** indicates that the mean differs significantly between Control and AM group (p < 0.001).
jkm-39-1-1f5.gif
Fig. 6
A. membranaceus increases the KGF expression in Hs68 fibroblast cells. Expression of KGF fluorescence by immunofluorescence staining (A) and protein level by Western blotting (B) in Hs68 fibroblast cells. Green, KGF. Blue, DAPI. Results are presented as mean ± S.E.M. *** indicates that the mean differs significantly between non-treated cells and AM-treated cells (p < 0.001).
jkm-39-1-1f6.gif

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