Anoectochilus roxburghii extract alleviates inflammation and restores skin barrier function in vitro and in vivo

Anoectochilus roxburghii (Wall.) Lindl. is a traditional medicinal herb used for clearing heat, cooling blood, and detoxifying. This study evaluated the anti-inflammatory and barrier-repair activities of anoectochilus roxburghii extract (ARE) using integrated in vitro and clinical approaches. ARE was prepared by water extraction and characterized by HPLC. Its mechanisms were investigated in UVB/capsaicin (CAP)/sodium dodecyl sulfate (SDS)-stimulated HaCaT cells, wound healing assays, and 3D epidermal models, then validated clinically. Quantitative analysis showed ARE contained 50.20% polysaccharides, 26.3% kinsenoside, 1.2% flavonoids, 2.5% polyphenols, 10.9% protein, and 2.8% aspartic acid. In HaCaT cells, ARE inhibited the NF-κB pathway and reduced TNF-α, IL-8, COX-2, and TRPV1 expression, while enhancing tight junction proteins (Claudin-1 and ZO-1). ARE promoted human primary keratinocyte migration. In 3D models, ARE upregulated filaggrin (FLG), loricrin (LOR), involucrin (IVL), and transglutaminase 1 (TGM1), while suppressing IL-1α and prostaglandin E2 (PGE2). Clinically, ARE formulations increased stratum corneum hydration, reduced transepidermal water loss (TEWL), and decreased skin redness. These findings demonstrate that ARE exerts multi-target anti-inflammatory and barrier-strengthening effects, supporting the traditional use of anoectochilus roxburghii for inflammatory skin conditions.