Integrated multi-omics reveals that Notoginsenoside R1 mitigates inflammation mediated by NLRP3 and metabolic disruption in muscle contusion

Muscle contusion triggers sterile inflammation and metabolic stress, in which macrophage-mediated NLRP3 inflammasome activation plays a critical role in secondary tissue injury and delayed repair. Panax notoginseng (SanQi) has long been used for traumatic injury and inflammation-related disorders, but its active constituents and immunopharmacological mechanisms in muscle contusion remain incompletely defined. Here, we investigated the protective effects of SanQi in a rat gastrocnemius contusion model and explored its underlying mechanisms using integrated transcriptomic, metabolomic, zebrafish, macrophage, molecular docking, molecular dynamics, and Western blot analyses. SanQi accelerated the resolution of muscle inflammation, reduced macrophage infiltration, and improved contusion-associated hemorheological abnormalities and hypercoagulability. Integrated multi-omics analyses showed that SanQi suppressed NLRP3 inflammasome-related inflammatory signaling and partially reversed metabolic alterations associated with muscle contusion, including succinate accumulation, TCA-cycle-associated metabolic changes, amino sugar and nucleotide sugar metabolism, and glycerophospholipid remodeling. Bioactivity-guided screening further identified Notoginsenoside R1 (NGR1) as a prioritized anti-inflammatory candidate constituent. In BMDMs, NGR1 selectively inhibited NLRP3 inflammasome activation and reduced caspase-1 cleavage and mature IL-1β release. Network pharmacology, molecular docking, molecular dynamics simulations, and Western blotting indicated that NGR1 interacts with the TLR4-MD2 complex and is associated with suppression of TLR4/NF-κB signaling. Collectively, our findings provide mechanistic insights into the pharmacological actions underlying the traditional "promoting blood circulation and removing blood stasis" effects of Panax notoginseng, with NGR1 identified as a prioritized bioactive saponin candidate contributing to its anti-inflammatory activity.