Virtual screening identifies specnuezhenide as a potential therapeutic agent for rosacea via KLK5/TLR4/NF-κB pathway inhibition and metabolic modulation.

In vivo animal study using a mouse model of rosacea induced by LL-37 injection, combined with in silico virtual screening and metabolomics analysis.

“Rosacea was induced in mice by intradermal LL-37 injection, followed by intraperitoneal administration of Spe.”

6–8-week-old Specific Pathogen Free BALB/c female mice.

“A total of 49 6–8-week-old Specific Pathogen Free BALB/c female mice were purchased from Henan Skebes Bio-technology Co., Ltd.”

49 mice divided into seven groups with 7 mice per group.

“The mice were randomly divided into seven groups (n = 7 mice per group): control (PBS), model (LL-37), doxycycline hydrochloride (DH, 30 mg/kg/d), LL-37 + low-dose Spe (20 mg/kg/d), LL-37 + medium-dose Spe (40 mg/kg/d), LL-37 + high-dose Spe (60 mg/kg/d), and LL-37 + high-dose Spe + KLK5 (60 mg/kg/d + 2 μg KLK5).”

Treatment and induction lasted for two consecutive days, with assessments performed 12 hours after the final injection.

“Spe was administered intraperitoneally at 20, 40, or 60 mg/kg/d for two consecutive days.”

Specnuezhenide (Spe) reduced LL-37-induced skin inflammation, decreased inflammatory cell and mast cell infiltration, and reduced MMP-9 expression and CD31 fluorescence intensity.

“Spe markedly reduced LL-37-induced skin inflammation, decreased inflammatory cell and mast cell infiltration, decreased MMP-9 expression, and reduced CD31 fluorescence intensity and CD4+ T cell recruitment.”

Spe inhibited the expression of pro-inflammatory cytokines (IL-1β, IL-6, and TNF-α) and blocked the activation of the TLR4/NF-κB pathway.

“The study demonstrated that Spe inhibited the expression of pro-inflammatory cytokines (IL-1β, IL-6, and TNF-α) in serum and suppressed activation of TLR2, MMP-9, KLK5, IL-1β, IL-6, and TNF-α genes.”

Metabolomics analysis identified 43 dysregulated metabolites in rosacea, 26 of which were normalized by Spe treatment, with pathway enrichment suggesting regulation of unsaturated fatty acid biosynthesis.

“Metabolomics identified 43 dysregulated metabolites in rosacea, 26 of which were normalized by Spe treatment.”

The study was conducted in a mouse model, and the metabolomic associations are statistical correlations requiring further functional validation to establish causal relevance.

“While the serum metabolomic data presented here reflect distal systemic consequences rather than the local drivers of lesional pathology, and the observed metabolite–pathway associations remain only statistical correlations, further targeted functional validation will be essential to establish their causal relevance and underlying mechanistic roles.”