Nutr Res Pract.  2023 Dec;17(6):1056-1069. 10.4162/nrp.2023.17.6.1056.

Extracts of Grifola frondosa inhibit the MAPK signaling pathways involved in keratinocyte inflammation and ameliorate atopic dermatitis

• Choi EJ
• Choi JK

• KMID: 2548864
• DOI: http://doi.org/10.4162/nrp.2023.17.6.1056

Abstract

BACKGROUND/OBJECTIVES
Grifola frondosa, commonly referred to as the maitake mushroom, has been studied extensively to explore its potential health benefits. However, its anti-inflammatory effects in skin disorders have not been sufficiently elucidated. This study aimed to elucidate the anti-inflammatory role of the ethanol extract of G. frondosa in atopic dermatitis (AD) using in vivo and in vitro models.
MATERIALS/METHODS
We investigated its impact on skin and spleen inflammatory responses in Dermatophagoides farinae extract (DFE)/1-chloro-2,4 dinitrochlorobenzene (DNCB)-induced AD-like skin lesions in a mouse model. Additionally, we determined the immunosuppressive response and mechanism of G. frondosa by inducing atopic-like immune reactions in keratinocytes through tumor necrosis factor (TNF)-α/interferon (IFN)-γ stimulation.
RESULTS
Our study revealed that G. frondosa ameliorates clinical symptoms in an ADlike mouse model. These effects contributed to the suppression of Th1, Th2, Th17, and Th22 immune responses in the skin and spleen, leading to protection against cutaneous inflammation. Furthermore, G. frondosa inhibited the production of antibodies immunoglobulin (Ig)E and IgG2a in the serum of AD mice. Importantly, the inhibitory effect of G. frondosa on inflammatory cytokines in TNF-α/IFN-γ-stimulated AD-like keratinocytes was associated with the suppression of MAPK (Mitogen Activated Protein Kinase) pathway activation.
CONCLUSIONS
Collectively, these findings highlight the potential of G. frondosa as a novel therapeutic agent for AD treatment and prevention.

Keyword

Grifola frondosa; atopic dermatitis; inflammation; cytokines; keratinocytes; mitogen-activated protein kinase

Figure

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  Fig. 1 FT-IR spectrum of the crude ethanolic extract of G. frondosa.

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  Fig. 2 G. frondosa treatment alleviates clinical symptoms in the AD-like skin lesion mouse model. (A) Study design for inducing AD-like skin lesions. Mice were grouped into four sets with 5 mice each. (B) 24 h post DNCB or DFE treatment, the ear thickness was gauged using a dial thickness instrument. (C) A photo representation of mouse ears from each typical group on the 28th day. (D) Microscope images of ear tissues stained with H&E or toluidine blue (at a 100× zoom, scale marker is 20 µm). (E) The thickness of both the epidermis and dermis was gauged from the H&E-stained tissue images. The count of invading mast cells was ascertained through toluidine blue staining. Data are presented as the mean ± SEM.Cera, ceramide; AD, atopic dermatitis; DNCB, 2,4-dinitrochlorobenzene; DFE, Dermatophagoides farina extract; H&E, hematoxylin and eosin.*P < 0.05; ****P < 0.0001 indicates significant reduction compared to AD.

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  Fig. 3 G. frondosa treatment reduces immunoglobulin levels in the blood and suppresses cytokines related to Th1, Th2, Th17, and Th22. G. frondosa treatment reduces immunoglobulin levels in the blood and suppresses cytokines related to Th1, Th2, Th17, and Th22. (A) Total serum IgE and IgG2a levels were measured by enzyme-linked immunosorbent assay. (B) Cytokine expression in the ears and spleen of mice exhibiting AD-like skin lesions. On the 28th day, the ears and spleen were removed and total RNA was extracted. The quantitative real-time polymerase chain reaction was conducted following the procedures outlined in the Methods section. Data are presented as the mean ± SEM.Ig, immunoglobulin; AD, atopic dermatitis; Cera, ceramide; ns, not significant.*P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001 indicates significant reduction compared to AD.

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  Fig. 4 G. frondosa suppresses chemokines and cytokines in keratinocytes. (A) The cytotoxic effects of G. frondosa on HaCaT cells were examined. The MTT assay was used to cell viability over a 24-h period. (B and C) HaCaT cells with or without G. frondosa or CsA pre-treatment for 1 h were stimulated with TNF-α (10 ng/mL) and IFN-γ (10 ng/mL) for 6 h to analyze gene expression. Pro-inflammatory cytokine gene expressions (TNF-α, IL-1β, and IL-6) and CCL17 were assessed by quantitative real-time polymerase chain reaction analysis (C). Data are presented as the mean ± SEM.TNF, tumor necrosis factor; IFN, interferon; CsA, Cyclosporine A; IL, interleukin.****P < 0.0001 indicates significant reduction compared to atopic dermatitis.

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  Fig. 5 G. frondosa downregulates the phosphorylation of MAPK signaling molecules such as p38 and ERK in keratinocytes. Inhibition of the p38, ERK, and JNK MAPKs pathways. Western blotting analysis of whole-cell extracts of HaCaT cells. Before being stimulated with TNF-α (10 ng/mL) and IFN-γ (10 ng/mL) for 30 min, cells were pre-treated with either G. frondosa (1 mg/mL) or CsA (1 mg/mL) for an hour.TNF, tumor necrosis factor; IFN, interferon; CsA, Cyclosporine A.

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