J Ethnopharmacol. 2026 Oct 8:122486. doi: 10.1016/j.jep.2026.122486. Online ahead of print.
ABSTRACT
ETHNOPHARMACOLOGICAL RELEVANCE: Hypoxia, a featured condition of high-altitude regions, contributes to the development of a wide range of cardiovascular and cerebrovascular diseases. However, the cardiovascular or cerebrovascular disease most strongly associated with hypoxia, along with its underlying molecular mechanisms and potential therapeutic targets, remains incompletely understood. Sanwei Tanxiang Powder (SWTX) is a classical Tibetan prescription that has long been used to improve blood circulation and alleviate hypoxia-related cardiovascular and cerebrovascular disorders. Licarin B is a bioactive constituent of SWTX with potential therapeutic value.
AIM OF THE STUDY: This study aimed to identify the cardiovascular and cerebrovascular disease most strongly associated with hypoxia, investigate its underlying molecular mechanisms, and explore the protective effects and molecular targets of licarin B against hypoxia-associated cerebrovascular injury.
MATERIALS AND METHODS: We employed an integrative strategy combining population-based epidemiological analysis, transcriptomics, network pharmacology, single-cell RNA sequencing, and experimental validation to systematically investigate hypoxia-associated cardiovascular and cerebrovascular diseases and their pathogenic mechanisms. Human cerebral microvascular endothelial cells (hCMEC/D3), were used to evaluate the effects of hypoxia and licarin B on oxidative stress, inflammatory responses, nitric oxide production, and endothelial function. The therapeutic efficacy of licarin B was further evaluated in a mouse model of acute ischemic stroke induced by middle cerebral artery occlusion (MCAO).
RESULTS: Epidemiological analysis identified ischemic stroke as the cardiovascular and cerebrovascular disease most strongly associated with hypoxia, as indicated by low hemoglobin levels. Multi-omics analyses revealed that hypoxia-associated stroke-related genes were predominantly enriched in inflammatory and oxidative stress pathways, with JUN identified as a key hypoxia-related stroke gene (HRSG). Single-cell transcriptomic analysis of murine brain tissue further demonstrated that JUN was relatively highly expressed in endothelial cells and positively associated with pro-inflammatory immune cell infiltration. In hCMEC/D3 cells exposed to hypoxia, JNK phosphorylation and JUN expression levels were markedly increased, accompanied by reactive oxygen species accumulation, impaired nitric oxide production, and enhanced expression of pro-inflammatory cytokines. Network pharmacology and molecular docking identified licarin B, a bioactive constituent of SWTX, as a candidate compound predicted to target JUN. In hypoxia-exposed hCMEC/D3 cells, licarin B significantly suppressed JUN expression, mitigated hypoxia-induced oxidative stress and inflammatory responses, and preserved endothelial barrier integrity, whereas JUN overexpression or JNK activation partially abrogated these protective effects. Furthermore, in a middle cerebral artery occlusion mouse model, licarin B administration reduced cerebral infarct volume, alleviated ischemic brain injury, suppressed the JNK/JUN signaling axis and pro-inflammatory cytokine expression, and improved neurological function.
CONCLUSIONS: This study identifies the hypoxia-JNK/JUN axis as a critical mediator of microvascular endothelial dysfunction during ischemic stroke and highlights licarin B, a bioactive constituent of SWTX, as a promising therapeutic candidate for hypoxia-associated cerebrovascular injury.
PMID:42849822 | DOI:10.1016/j.jep.2026.122486