Zhen-Wu-Tang ameliorates renal fibrosis and tissue stiffness in a murine lupus model, an effect associated with the suppression of fibroblast-to-myofibroblast transition.

Abstract

ETHNOPHARMACOLOGICAL RELEVANCE: Pathological extracellular matrix (ECM) deposition is a hallmark of lupus nephritis (LN) progression. Zhen-Wu-Tang (ZWT), a classic traditional formula, is clinically used for treating LN and has demonstrated anti-fibrotic properties. OBJECTIVE: This study aimed to evaluate the therapeutic effects of ZWT on ECM dysregulation in LN and its mechanisms of action. METHODS: Spontaneous LN models (MRL/lpr mice) and TGF-β-induced fibroblasts were used. Kidney transcriptomes were analyzed using RNA-seq. Renal ECM deposition and tissue stiffness were assessed via immunohistochemistry, PCR, and atomic force microscopy. Key mechanosensitive molecules (YAP, TAZ, Piezo1, ROCK1/2) were examined by RT-PCR and immunofluorescence. ZWT-containing serum was applied in vitro to assess fibroblast activation and ECM secretion, while ECM-related pathways (TGF-β-Smad2/3, NF-κB, PI3K/AKT, JAK2/STAT3) were detected via immunoblotting. RESULTS: ZWT treatment significantly ameliorated renal fibrosis in LN mice by suppressing fibronectin, collagen I, and collagen III deposition. Transcriptomics identified 66 ECM-related genes modulated by ZWT. ZWT also reduced renal stiffness and downregulated mechanosensitive signaling components. Finally, ZWT inhibited EMT and ECM overproduction by fibroblasts via concurrently suppressing Smad2/3, NF-κB, PI3K/AKT, and JAK2/STAT3 pathways. CONCLUSION: In summary, ZWT ameliorates renal fibrosis and renal stiffness in a murine lupus model, primarily by suppressing fibroblast activation via multi-pathway inhibition. These findings suggest its potential as a multi-target therapeutic agent for LN.