Essential oil from Angelica sinensis Radix attenuates ischemic stroke via the cGMP/PKG/MAPK pathway: insights from network pharmacology and molecular mechanism evaluation.

ETHNOPHARMACOLOGICAL RELEVANCE: As a traditional herbal remedy, the root of Angelica sinensis has long been used to treat headaches, strokes, and dementia. As the primary active ingredient in A. sinensis, A. sinensis essential oil (ASEO) also plays a significant role in the therapy of stroke. Although ASEO has shown clear therapeutic effects, its specific mechanism of preventing ischemic stroke still needs to be further clarified.

AIM OF THE STUDY: Through network pharmacological analysis combined with in vitro and in vivo experimental verification, this study investigated the protective effects of ASEO against stroke and elucidated the underlying molecular regulatory mechanisms.

MATERIALS AND

METHODS

Gas chromatography-mass spectrometry (GC-MS) was used to analyze the chemical composition of ASEO. Middle cerebral artery occlusion/reperfusion (MCAO/R) rats were administered different doses of ASEO to evaluate its therapeutic effects. The therapeutic effects were evaluated using behavioral, histopathological, and immunofluorescence assays. In addition, using network pharmacology techniques, the potential molecular mechanisms underlying the treatment of ischemic stroke with ASEO have been preliminarily elucidated. HT22 cells treated with oxygen-glucose deprivation/reoxygenation (OGD/R) were used as an in vitro model. Subsequently, the key signal transduction pathways involved in the therapeutic effects of ASEO against ischemic stroke were further validated using multiple experimental approaches, including enzyme-linked immunosorbent assay (ELISA), flow cytometry, Western blotting, pathway-specific pharmacological inhibition and PKG-specific siRNA knockdown.

RESULTS

Twenty-seven compounds were identified from ASEO by GC-MS. In vivo experiments demonstrated that ASEO treatment exerted significant protective effects on MCAO/R rats by improving impaired neurological function, reducing cerebral infarct volume, alleviating neuronal apoptosis, and modulating the expression of apoptotic proteins. Mechanistically, network pharmacology studies suggested that the protective mechanism of ASEO may involve the cGMP/PKG and MAPK signaling pathways. Subsequent experiments confirmed that ASEO inhibited neuronal apoptosis via the cGMP/PKG/MAPK axis both in vivo and in vitro. Importantly, both pharmacological inhibition and siRNA-mediated genetic knockdown of PKG significantly attenuated the protective effects of ASEO against OGD/R injury in HT22 cells.

CONCLUSION

ASEO exerts its anti-ischemic stroke effect by attenuating apoptosis via the cGMP/PKG/MAPK signaling pathway, providing a scientific foundation for its clinical application in ischemic stroke therapy.