Yin-Hua Li-Shi Formula Exerts Anti-Atopic Dermatitis Effects Through Luteolin Targeting AKT/MAPK-IL-6/IL-17 Axis

Xuemin Wang,1,* Ning Jia,2,* Hyewon Joo,3,* Shuxin Wang,4 Caiyun Zhang,2 Qilong Chen,2 Wencheng Jiang1 1Department of Traditional Chinese Medicine Dermatology, Shanghai Skin Disease Hospital, School of Medicine, Tongji University, Shanghai, 200443, People’s Republic of China; 2Central Laboratory, Shanghai Skin Disease Hospital, School of Medicine, Tongji University, Shanghai, 200443, People’s Republic of China; 3Allergic Dermatoses Clinical Center, Shanghai Skin Disease Hospital, Tongji University School of Medicine, Shanghai, 200443, People’s Republic of China; 4Guang’anmen Hospital, China Academy of Chinese Medical Sciences, Beijing, 100053, People’s Republic of China*These authors contributed equally to this workCorrespondence: Wencheng Jiang, Shanghai Skin Disease Hospital, Tongji University School of Medicine, Shanghai, 200443, People’s Republic of China, Email [email protected] Qilong Chen, Shanghai Skin Disease Hospital, Tongji University School of Medicine, Shanghai, 200443, People’s Republic of China, Email [email protected]: Yin-Hua Li-Shi Formula (YHLS), a medically authorized traditional Chinese medicine (TCM) formulation for atopic dermatitis (AD), has a 30-year clinical application history in China, yet has not had its precise therapeutic mechanisms fully elucidated to date.Methods: Chemical profiling of YHLS was conducted using high-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Network pharmacology predicted YHLS’s active ingredients, therapeutic targets, and anti-atopic dermatitis (AD) pathways, with molecular docking validating component-target binding affinity. By intersecting network pharmacology/molecular docking results with mass spectrometry profiles, the key bioactive component was identified. For in vivo validation, an AD-like model was generated via topical MC903 application in C57BL/6 mice. Dermatitis severity, ear thickness, H&E staining, ELISA, RT-qPCR, and Western blot were performed to assess YHLS’s efficacy and underlying mechanisms.Results: Luteolin, the major bioactive component of YHLS targeting AD, was identified by intersecting UPLC-MS/MS metabolite profiles with network pharmacology and molecular docking results. Network pharmacology revealed 102 common targets between YHLS and AD, with PPI/KEGG analyses confirming MAPK1 and AKT1 as core inflammatory targets. Molecular docking assays demonstrated that luteolin binds tightly to MAPK1 (− 8.2 kcal/mol) and AKT1 (− 9.7 kcal/mol). In MC903-induced AD-like mice, YHLS alleviated symptoms dose-dependently (reduced severity scores, ear thickness, hyperkeratosis, and inflammatory infiltration; all P< 0.05). High-dose YHLS decreased serum IgE (P< 0.001) and inhibited AKT1/MAPK1 phosphorylation. RT-qPCR showed YHLS downregulated IL-6 (high-dose P< 0.05) and potently suppressed IL-17 (all doses P< 0.0001, dose-dependent), with TNF-α also reduced.Conclusion: Luteolin is the major bioactive component of Yin-Hua Li-Shi Formula (YHLS) that mediates its anti-atopic dermatitis effects by specifically targeting the AKT/MAPK-IL-6/IL-17 axis. Keywords: atopic dermatitis, AD, Yin-Hua Li-Shi formula, YHLS, network pharmacology, molecular docking, animal experiment, inflammation, AKT/MAPK pathway