Integrating network pharmacology and pharmacological evaluation for investigating the active ingredient of Chinese herbal formula Zhuhuang Granule in psoriasis.

OBJECTIVE: Psoriasis is a chronic, relapsing inflammatory skin disease associated with multiple factors, including genetics and environmental influences. The etiology and pathogenesis of this disease are not yet fully elucidated. Zhuhuang Granules (ZG) can improve psoriasis; however, their related molecular mechanism is not well defined. Therefore, this study aims to investigate the primary components and their molecular mechanisms for treating psoriasis through bioinformatics analysis and molecular biology experiments.

METHODS: The differentially expressed genes (DEGs) in psoriatic lesion areas and normal skin in the GEO datasets GSE14905 and GSE13355 were analyzed. Pathway enrichment analysis of DEGs was performed using Metascape and via the ClusterProfiler package. Protein-protein interaction (PPI) network analysis of DEGs was used using the STRING database. Target genes for the main active ingredients of ZG were predicted using SymMap. Cytoscape was used to find the core target genes of the main components of ZG. In the in-vitro experiments, HaCaT cells, upon treatment with IL-22 and Cortex Phellodendri extracts (HB extracts), were used for the measurement of cell viability, the levels of inflammatory factors (IL-17A, IL-1β, and TNF-α), as well as the expression levels of CTNNBIP1 and the Wnt pathway-related proteins by CCK-8 assay, ELISA, and western blot. In the in vivo experiments, the effects of HB extracts and CTNNBIP1 knockdown on epidermal abnormal proliferation and inflammatory factor secretion in mice were examined using HE staining, IHC staining, ELISA, and western blot after induction of the mouse models of psoriasis with imiquimod (IMQ).

RESULTS: The pathway enrichment results indicated a close correlation between DEGs of GSE14905 and GSE13355 and the pathological process of psoriasis. Cytoscape results revealed that Cortex Phellodendri was located in the core of ZG and targeted the most genes (n=20). CTNNBIP1 was selected for subsequent experiments, in combination with the group's previous studies. The in vitro experiments suggested that HB extracts affected HaCaT cell proliferation and inflammatory factor secretion by promoting CTNNBIP1 expression. The in-vivo experiments demonstrated that HB extracts significantly alleviated the symptoms of psoriasis in mice, reduced ki-67 and IL-17A expression in skin tissues, and upregulated CTNNBIP1 expression. Besides, the knockdown of CTNNBIP1 in the skin partially reversed the effect of HB extracts.

CONCLUSION: This research underlines that HB extracts suppress the Wnt pathway activation and keratinocyte proliferation and inflammation through the upregulation of CTNNBIP1, ultimately improving psoriasis.