Kushe Tincture Ameliorates DNCB-Induced Atopic Dermatitis by Affecting NF-Kb/JAK-STAT3 Pathway: Bioinformatics Analysis and Animal Experiment Verification.

BACKGROUND

Kushe tincture (KSD), an optimized in-hospital preparation of Dongzhimen Hospital of Beijing University of Traditional Chinese Medicine, has been clinically used as an anti-inflammatory and anti-itching topical therapeutic drug for the treatment of eczema, atopic dermatitis (AD) and so on for decades. However, the potential therapeutic mechanisms remained unexplored.

OBJECTIVE

This study aimed to explore the mechanisms underlying the therapeutic effects of KSD against DNCB-induced AD through the combination of single-cell RNA sequencing (scRNA-seq), spatial transcriptomics (ST), and animal experiments.

METHODS

Firstly, we collected the active components and targets of KSD from the Traditional Chinese Medicine Systems Pharmacology (TCMSP) and obtained AD-associated targets from the Gene Expression Omnibus (GEO) database. Protein-protein interaction (PPI) networks were constructed using Cytoscape with Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses. Secondly, core targets were screened and visualized by scRNA-seq and ST technology. Finally, we adopted DNCB-induced AD-like mouse model to observe the therapeutic effect of KSD through lesion scoring, morphological staining, serological tests, skin barrier function measurements, flow cytometry, and Western blotting.

RESULTS

Forty-six common drug-disease targets were obtained and top 2 ranked targets were chosen, namely NF-kB1 and STAT3. Combined scRNA-seq and ST suggested that NF-KB and STAT3 had higher and wider expression in most subpopulations, especially in monocytes and CD4+ T-cells. Animal experiments showed that KSD treatment improved the visible symptoms and skin barrier function and decreased mast cell infiltration, IgE levels, Th17 cells, and expression of NF-kB, JAK2, and STAT3.

CONCLUSION

KSD has therapeutic effects on DNCB-induced AD-like skin lesions, and it has effects on NF-kB and JAK-STAT3 proteins, which may be possible targets.