Anti-Atopic Effect of Scutellaria baicalensis and Raphanus sativus on Atopic Dermatitis-like Lesions in Mice by Experimental Verification and Compound-Target Prediction

Abstract

Scutellaria baicalensis Georgi and Raphanus Sativus Linne herbal mixture (SRE) is a Chinese herbal medicine. In this study, we aimed to evaluate the therapeutic efficacy of SRE as an active ingredient for 2,4-dinitrochlorobenzene (DNCB)-induced atopic dermatitis (AD) and to predict the underlying therapeutic mechanisms and involved pathways using network pharmacological analysis. Treatment with SRE accelerated the development of AD-like lesions, improving thickness and edema of the epidermis. Moreover, administering the SRE to AD-like mice suppressed immunoglobulin E and interleukin-4 cytokine and reduced T lymphocyte differentiation. In silico, network analysis was used to predict the exact genes, proteins, and pathways responsible for the therapeutic effect of the SRE against DNCB-induced AD. These results indicated that the SRE exerted protective effects on the DNCB-induced AD-like model by attenuating histopathological changes and suppressing the levels of inflammatory mediators. Therefore, the SRE can potentially be a new remedy for improving AD and other inflammatory diseases and predicting the intracellular signaling pathways and target genes involved. This therapeutic effect of the SRE on AD can be used to treat DNCB-induced AD and its associated symptoms.

Keywords: Raphanus sativus; Scutellaria baicalensis; T lymphocyte differentiation; atopic dermatitis; network pharmacology; skin.

Figure 1

Effects of SRE on atopic dermatitis-like symptoms in hairless mice. (A) Schematics of experiment design. (B) After 30 days, images of skin lesions from the groups were taken on the last day of treatment. (C) Value of transepidermal water loss (TEWL). (D) Level of skin hydration. (E) Spleen weight in hairless mice (n = 7). All data are presented as mean ± SEs; * p < 0.05 compared with the AD group, and # p < 0.05 compared with the sham group—AD: atopic dermatitis; Dex: dexamethasone; DNCB: 2,4-dinitrochlorobenzene.

Figure 2

Effects of SRE on atopic dermatitis-like histological changes in hairless mice. (A) H&E staining of the skin lesion; scale bar = 200 μm (B) Epidermal thickness was analyzed in H&E-stained sections (n = 3). All data are presented as mean ± SEs; * p < 0.05 compared with the AD group, and # p < 0.05 compared with the sham group. AD: atopic dermatitis; Dex: dexamethasone.

Figure 3

Effects of SRE on lymphocyte cells—IgE and IL-4 levels in atopic dermatitis-like hairless mice. (A) Dot plot of representative fluorescence-activated cell sorting analysis in the spleen. (B) The percentage of GATA3+ and CD25+. (C) Level of IgE in serum. (D) Level of IL-4 in hairless mice serum (n = 7). All data are presented as mean ± SEs; * p < 0.05 compared with the AD group, and # p < 0.05 compared with the sham group. AD: atopic dermatitis; IgE: immunoglobulin E; IL-4: interleukin-4; CD: cluster of differentiation; GATA: GATA binding protein 3; Dex: dexamethasone.

Figure 4

Target and potential genes of SRE. (A) HPLC-DAD chromatogram of the sample (254 nm) (B) Network analysis of herbs–small molecules–genes; the green hexagon is an herb, the pink oval is the small molecule, and the cyan rectangle is a gene. (C) Protein–protein interactions (PPIs): the core gene is the larger size of the circle and the more orange color in SRE.

Figure 5

Pathway analysis of SRE: (A) KEGG classification of pathways. (B) Rank according to the p-value. (C) Pathways (triangle orange color) and their corresponding genes (rectangle cyan color) for SRE.