Objectives: Saffron, a traditional Chinese medicine, is renowned for its pharmacological effects in promoting blood circulation, resolving blood stasis, regulating menstruation, detoxification, and alleviating mental disturbances. Trans-crocetin, its principal bioactive component, exhibits significant anti-hypoxic activity. The clinical development and therapeutic efficacy of trans-crocetin are limited by its instability, poor solubility, and low bioavailability. Conversion of trans-crocetin into trans-sodium crocetinate (TSC) enhances its solubility, stability, and bioavailability, thereby amplifying its anti-hypoxic potential. Methods: This study integrates network pharmacology with in vivo and in vitro validation to elucidate the molecular targets and mechanisms underlying TSC’s therapeutic effects against high-altitude acute lung injury (HALI), aiming to identify novel treatment strategies. Results: TSC effectively reversed hypoxia-induced biochemical abnormalities, ameliorated lung histopathological damage, and suppressed systemic inflammation and oxidative stress in HALI rats. In vitro, TSC mitigated CoCl₂-induced hypoxia injury in human pulmonary microvascular endothelial cells (HPMECs) by reducing inflammatory cytokines, oxidative stress, and ROS accumulation while restoring mitochondrial membrane potential. Network pharmacology and pathway analysis revealed that TSC primarily targets the EGFR/PI3K/AKT/NF-κB signaling axis. Molecular docking and dynamics simulations demonstrated stable binding interactions between TSC and key components of this pathway. ELISA and RT-qPCR confirmed that TSC significantly downregulated the expression of EGFR, PI3K, AKT, NF-κB, and their associated mRNAs. Conclusions: TSC alleviates high-altitude hypoxia-induced lung injury by inhibiting the EGFR/PI3K/AKT/NF-κB signaling pathway, thereby attenuating inflammatory responses, oxidative stress, and restoring mitochondrial function. These findings highlight TSC as a promising therapeutic agent for HALI.

Hydrogen sulfide (H₂S) is an important endogenous gaseous signaling molecule that regulates diverse physiological and pathological processes through protein persulfidation. As a central regulator of inflammation and immune responses, NF-κB signaling is precisely controlled by post-translational modifications, and its dysregulation contributes to various inflammatory diseases. Recent studies reveal that H₂S-mediated persulfidation is a key mechanism for fine-tuning NF-κB activity by targeting critical components, including p65, IKKβ, IκBα, and upstream regulators. Through site-specific S-sulfhydration, H₂S modulates IKK activation, IκBα degradation, and p65 nuclear translocation, thereby limiting excessive NF-κB activation and inflammatory cytokine production. This review provides an integrated view of how endogenous H₂S production and persulfidation-dependent signaling regulate inflammatory responses. Rather than simply summarizing individual pathways, we focus on the molecular mechanisms underlying H₂S-mediated regulation of NF-κB-associated networks, including TLR4/NF-κB, PI3K/Akt/NF-κB, and MAPK/NF-κB pathways, and highlight its roles in oxidative stress, apoptosis, pyroptosis, and tissue repair. We further discuss current challenges in identifying persulfidation sites, understanding endogenous H₂S regulation, and improving detection technologies. By proposing H₂S as a precision modulator of inflammatory signaling, this review provides new insights into H₂S biology and highlights future opportunities for developing targeted H₂S-based therapeutic strategies.