Integrative multi‑omics and experimental validation reveal that Duhuo Jisheng Decoction alleviates IVDD by inhibiting...

Intervertebral disc degeneration (IVDD) is a chronic and progressive condition with limited therapeutic options. Duhuo Jisheng Decoction (DHJSD), a traditional Chinese medicine formula, is clinically used to alleviate IVDD, but its underlying mechanisms remain unclear.

This study aims to investigate the active components and molecular mechanisms of DHJSD in treating IVDD, with a focus on the p38MAPK signaling pathway and mitochondrial homeostasis.

Bioinformatic analyses, including transcriptomic profiling, single-cell RNA sequencing, and network pharmacology, were performed to identify active components and potential targets. The findings were validated using in vitro (LPS-induced degenerated nucleus pulposus cells) and in vivo (rat tail puncture-induced IVDD model) experiments.

Transcriptomic analysis revealed 2202 differentially expressed genes, and 10 hub genes (e.g., TP53, AKT1, TNF) were identified. Single-cell analysis identified 12 cell types in degenerated NP tissues, with immune cells enriched in early degeneration and fibrotic NP cells in advanced stages. Network pharmacology screened 28 hub targets, with 11 core targets (e.g., IL-1β, TNF, CASP3) enriched in MAPK and inflammation-related pathways. In vivo, DHJSD treatment significantly improved disc height (DR imaging) and water content (MRI), restored histological structure (HE and Safranin-O staining), and reduced serum TNF-α and IL-1β levels (*p < 0.05). In vitro, DHJSD-containing serum (medium dose, 48 h) significantly reversed LPS-induced decreases in cell proliferation (CCK-8), upregulated Col II and Agg expression, and downregulated TNF-α, IL-1β, MMP-2, CASP3, and p38MAPK/JNK phosphorylation (*p < 0.05). DHJSD also partially restored mitochondrial membrane potential, indicating improved mitophagy.

DHJSD alleviates IVDD by suppressing inflammation, ECM degradation, and NP cell apoptosis, likely via regulating mitophagy through the p38MAPK signaling pathway. Collectively, this work not only establishes DHJSD as a modulator of the p38MAPK-mitophagy axis but also offers a mechanistic paradigm that shifts the understanding of TCM-based IVDD therapy from broad efficacy to pathway-specific intervention.