An integrated approach combining computational analyses and experimental validation deciphers the mechanism and active substance basis of Huazhuo Ningfu decoction in treating psoriasis.

Abstract

This study aims to elucidate the mechanism of HZD in treating psoriasis and identify its active components through a comprehensive approach combining tandem mass tag (TMT)-based quantitative proteomics with machine learning and molecular docking analysis. An integrated strategy was employed, combining tandem mass tag (TMT)-based quantitative proteomics and machine learning to identify critical targets in an imiquimod (IMQ)-induced mouse model of psoriasis. Key findings were then experimentally validated using the human immortalized keratinocyte (HaCaT) cell model and HZD-treated mice through techniques including 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), quantitative real-time PCR (qPCR), Western blot (WB), and immunofluorescence (IF). Furthermore, molecular docking and dynamics simulations were utilized to investigate interactions between HZD's bioactive components and the identified targets. Our findings demonstrate that HZD alleviates psoriasis primarily by inhibiting the Notch signaling pathway and upregulating keratin 77 (KRT77), thereby normalizing keratinocyte function. The identification of nuciferine as a key multi-target constituent, along with its structural analogs, provides a pharmacological basis for HZD's efficacy and reveals promising leads for novel psoriasis therapeutics.