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. 2022 Oct;10(20):1104.
doi: 10.21037/atm-22-4423.

Cucurbitacin promotes hair growth in mice by inhibiting the expression of fibroblast growth factor 18

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Cucurbitacin promotes hair growth in mice by inhibiting the expression of fibroblast growth factor 18

Keke Guo et al. Ann Transl Med. 2022 Oct.

Abstract

Background: The inhibition of fibroblast growth factor 18 (FGF18) promotes the transition of hair follicles (HFs) from the telogen phase to the anagen phase. Cucurbitacin has been shown to have a good effect in promoting hair cell growth. This study explored the potential effect of cucurbitacin on hair growth and its effect on FGF18 expression in mice.

Methods: Male C57BL/6J mice were randomly divided into the following two groups: (I) the vehicle group; and (II) the cucurbitacin group. Matrix cream and cucurbitacin cream were applied to the depilated skin on the back of the vehicle group mice and the cucurbitacin group mice, respectively. On days 3, 6, 9, 12, 15, and 18, the hair growth in the depilated dorsal skin of the mice was recorded with a digital camera and a HF detector, and the HF cycle status of the mice was observed by hematoxylin and eosin (H&E) staining. In addition, the level of FGF18 messenger ribonucleic acid (mRNA) in the dorsal skin was measured on days 15 and 18 by quantitative real-time polymerase chain reaction (qRT-PCR), while the level of FGF18 protein was measured by western blot and immunofluorescence staining.

Results: The dorsal skin to which the cucurbitacin cream was applied began to darken on day 6 and grew hairs on day 9, which was 3 days earlier than the dorsal skin to which the matrix cream was applied. The H&E staining revealed a transition from the telogen phase to the anagen phase 3 days earlier for the cucurbitacin cream-treated skin than the matrix cream-treated skin. In addition, the skin treated with cucurbitacin cream also showed a significant decrease in FGF18 mRNA as seen by qRT-PCR, and reduced FGF18 protein levels as detected by western blot and immunofluorescence staining compared to the skin treated with matrix cream only.

Conclusions: Cucurbitacin significantly reduced the levels of FGF18 mRNA and protein in the dorsal skin of mice to accelerate the HFs to enter the anagen phase earlier, thereby promoting the regeneration of hair. Thus, cucurbitacin can be considered a new and valuable agent for the development of anti-hair loss products.

Keywords: Cucurbitacin; fibroblast growth factor 18 (FGF18); hair follicle cycle.

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Conflict of interest statement

Conflicts of Interest: All authors have completed the ICMJE uniform disclosure form (available at https://atm.amegroups.com/article/view/10.21037/atm-22-4423/coif). All authors report that this work was supported by Infinitus (China) Co., Ltd. The authors have no other conflicts of interest to declare.

Figures

Figure 1
Figure 1
Effect of cucurbitacin on hair regeneration in the depilated back skin of mice. (A) Diagram depicting the experimental design. (B) Gross observation images of the depilated back skin following the application of matrix cream or cucurbitacin cream. The images were taken on different days as indicated. (C) Changes in the morphology of the HFs in the depilated skin after the application of matrix cream or cucurbitacin cream. The images were taken with a HF detector at 200× magnification and on different days as indicated. (D) H&E staining of dorsal skin tissues after the application of matrix cream or cucurbitacin cream. Images were taken on different days as indicated. Scale bars: 10 μm. HF, hair follicle; H&E, hematoxylin and eosin.
Figure 2
Figure 2
Effect of cucurbitacin on the expression of FGF18 in the depilated back skin of mice. (A) qRT-PCR analysis of FGF18 mRNA expression levels in the skin on days 15 and 18 after applying matrix cream or cucurbitacin cream. (B) Western blot analysis of FGF18 protein expression levels in the skin on days 15 and 18 after applying matrix cream or cucurbitacin cream. The plot below the blot shows the quantitation of the FGF18 protein in the blot as determined by grayscale densitometry. The results were normalized to α-tubulin expression. (C) Immunofluorescence staining of FGF18 protein expression levels in the skin on days 15 and 18 after applying matrix cream or cucurbitacin cream. Green fluorescence represents FGF18 protein. All tissue slices were counterstained with DAPI (blue). Scale bars: 50 μm. The plot besides the image compares the level of FGF18 protein between the two different treatments. Data in the plots are mean ± SEM from 3 experiments, ‘*’ and ‘**’ indicate that the results of the mice that received the cucurbitacin cream treatment differed significantly to the results of the mice that received the matrix cream treatment at the P<0.05 and P<0.01 levels, respectively. FGF18, fibroblast growth factor 18; qRT-PCR, quantitative real-time polymerase chain reaction; DAPI, 4, 6-diamino-2-phenyl indole; SEM, standard error of mean.

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