Targeting TSLP and NFkB in dermatitis using computational and experimental validation of luteolin, quercetin, and cetirizine allopolyherbal combination

Atopic dermatitis (AD) is a rapidly escalating global health concern, affecting up to 20% of children and 10% of adults, with current therapies often proving inadequate due to limited efficacy, adverse effects, and lack of multi-target action. This study addresses a significant knowledge gap by investigating a novel, multi-targeted combination strategy involving luteolin, quercetin, and cetirizine to modulate key molecular mechanisms underlying AD, namely oxidative stress, inflammatory signaling, and histamine-mediated immune response. We hypothesized that the combination of these three compounds would exert synergistic anti-inflammatory and antioxidant effects, surpassing their individual performances using computational studies and experimental validation. In silico molecular docking revealed strong binding affinities of luteolin and quercetin with thymic stromal lymphopoietin (TSLP) at -6.2 and -6.4 kcal/mol, respectively, while cetirizine showed the highest affinity for histamine H1 receptor (-9.1 kcal/mol). Molecular dynamics simulations over 100 ns confirmed stable ligand-protein interactions. Network pharmacology analysis further suggested these compounds collectively modulate key signalling pathways in AD pathogenesis. In vitro assays validated these findings, with the combination showing superior DPPH radical scavenging (IC50: 1.5394 ± 0.056 µg/mL), NO• assay (84.1 ± 1.68%), and anti-inflammatory activity via albumin denaturation (88.2 ± 1.76%). A synergy value of 35.37±1.52% confirmed a positive interaction. These results support the therapeutic promise of this flavanoid-antihistamine combination as a multi-targeted strategy for AD management.