Pinggan Yuyin Qingre formula ameliorates meibomian gland dysfunction through the interleukin-17/nuclear factor kappa B pathway based on network pharmacology and experimental validation.

OBJECTIVE: validation. METHODS: A mouse model of MGD was induced using the stearoyl-coenzyme a desaturase 1 inhibitor, followed by PGYYQR treatment for 2 weeks. MGD sign scoring, hematoxylin and eosin (HE) staining, oil red o (ORO) staining, and serum inflammatory cytokine analysis were conducted to assess the effects of PGYYQR on meibomian gland (MG) function, histopathology, and associated inflammation. Network pharmacology was employed to identify the active compounds and potential targets of PGYYQR. Molecular mechanisms were further investigated using Western blotting, reverse transcription quantitative real-time polymerase chain reaction, and reactive oxygen species (ROS) assays. RESULTS: PGYYQR treatment significantly reduced the scores of MG orifice obstruction and meibum quality in MGD mice. HE and ORO staining further demonstrated that PGYYQR ameliorated glandular damage and lipid dysfunction. Enzyme-linked immunosorbent assay results revealed that PGYYQR markedly decreased the serum levels of key inflammatory cytokines, including interleukin (IL)-1β, IL-6, and tumor necrosis factor-α. Network pharmacology identified 162 active compounds and 598 target genes in PGYYQR. Among these, IL-6, IL-1β, matrix metalloproteinase-9, and C-X-C motif chemokine ligand 8 were recognized as core targets related to MGD and were mainly enriched in the IL-17/ nuclear factor kappa B (NF-κB) signaling pathway. Further molecular analyses confirmed that PGYYQR significantly inhibited the IL-17/NF-κB axis by downregulating IL-17 expression and reducing phos-phorylated NF-κB p65 levels at both the protein and mRNA levels in MG tissues. PGYYQR also effectively reduced ROS levels in the conjunctival tissues of MGD mice. CONCLUSION: PGYYQR effectively improves MG function and preserves local tissue morphology in MGD model mice, primarily by suppressing the inflammatory response through coordinated modulation of the IL-17/NF-κB signaling pathway and oxidative stress.