Peitu Qingxin formula ameliorates atopic dermatitis by targeting the multi-omics-informed miR-30a/GAPR-1 axis in sphingolipid metabolism.

BACKGROUND

Dysregulated sphingolipid metabolism plays a key role in atopic dermatitis (AD), yet therapeutic options targeting this pathway remain limited. The traditional formula Peitu Qingxin (PTQX) shows clinical efficacy, but its molecular mechanism is largely undefined.

PURPOSE: To identify a novel mechanism of the PTQX formula in AD sphingolipid metabolism and to validate the miR-30a/GAPR-1 axis as a therapeutic target.

METHODS

Using Mendelian randomization, clinical proteomics, and dual-luciferase reporter assays, we identified miR-30a as a direct upstream regulator of GAPR-1. Gain-of-function studies in AD-like 3D skin equivalents validated this axis. The effective dose of PTQX was determined in NC/Nga mice induced with 2,4-dinitrochlorobenzene. Molecular docking predicted the binding of PTQX components to GAPR-1. Adenovirus-mediated GAPR-1 silencing was performed in AD mice, and targeted lipidomics was used for detection. Finally, PTQX efficacy was evaluated in miR-30a knockout mice by assessing dermatitis severity, inflammatory cytokines/proteins, proteomics, and untargeted metabolomics.

RESULTS

We found that miR-30a directly targets GAPR-1 and may exert a protective role in AD. In AD-like 3D skin equivalents, miR-30a overexpression downregulated GAPR-1, restored barrier proteins, and suppressed IL-8. Medium-dose PTQX treatment upregulated miR-30a and downregulated GAPR-1, concomitantly with amelioration of dermatitis. PTQX components were predicted to have strong binding affinity to GAPR-1 in silico. Adenovirus-mediated GAPR-1 silencing alleviated AD-like symptoms and restored sphingolipid metabolites. In miR-30a knockout mice, GAPR-1 expression was elevated. PTQX treatment suppressed serum levels of KC, IL-5, IL-12p70, IL-10, G-CSF, and TNF-α, reduced skin protein levels of NLRP3 and TLR2, and restored CYP1A1 expression. Multi-omics analyses further revealed that sphingolipid dysregulation is a key downstream mechanism. PTQX reversed the downregulation of key sphingolipid enzymes GBA and ENPP7, as well as 7 sphingolipid metabolites.

CONCLUSION

Targeting the miR-30a/GAPR-1 axis by PTQX represents a novel therapeutic approach for treating sphingolipid dysregulation in AD.