Data Sheet 1_Mechanistic study on components of Angelica Pubescens-Taxillus Chinensis in improving compression-induced intervertebral disc degeneration via piezo1/NF-κB signaling axis.pdf

Additionally, the impact of CBCM on the Piezo1 and NF-κB pathways was analyzed through Western blotting, qRT-PCR, and immunofluorescence double staining. The underlying mechanism was confirmed by rescue experiments employing a Piezo1 agonist and antagonist, as well as p65 knockdown and overexpression. In vivo, a rat tail disc compression-induced IVDD model was established.

The CBCM was administered intraperitoneally. IVDD was evaluated via X-ray, MRI, and histology. Molecular mechanisms underlying senescence, ECM degradation, and the Piezo1/NF-κB axis were analyzed by ELISA and Western blotting.

Molecular docking was performed to further validate plausible interactions of CBN, OST, and QR with Piezo1.</p>Results<p>Compression effectively induced NP cell senescence and activated Piezo1 expression and NF-κB signaling. The CBCM concentration ratio that optimally suppressed senescence markers was quercetin at 50 μM, osthole at 6.25 μM, and columbianadin at 12.5 μM. Mechanistically, the optimized CBCM inhibited Piezo1 expression, NF-κB activation, and p65 nuclear translocation.

Both p65 knockdown and Piezo1 inhibition mimicked CBCM’s anti-senescent effects, whereas p65 overexpression or Piezo1 agonism reversed these protective effects. These in vitro findings were corroborated in a rat tail compression model of IVDD. Molecular docking offered computational evidence supporting the potential direct binding of CBN, OST, and QR with Piezo1 and proteins involved in the NF-κB signaling pathway.</p>Conclusion<p>These findings suggest that the optimal Angelica Pubescens-Taxillus Chinensis CBCM attenuates the compression-induced senescence-associated phenotype of NP cells in vitro and in a rat tail model by modulating Piezo1-mediated NF-κB signaling.</p>