Alpinia oxyphylla Miq. promotes angiogenesis and preserves neurons in subacute ischemic stroke in rats: possible involvement of the HIF-1α/Nrf2/VEGF-A-mediated signaling pathway.

ETHNOPHARMACOLOGICAL RELEVANCE Alpinia oxyphylla Miq., referred to in Chinese as Yi Zhi Ren (YZR), is a traditional Chinese herbal medicine long used to treat stroke. However, the effects of YZR extract on angiogenesis in subacute ischemic stroke remain unclear.

Aim

OF THE STUDY The effects of YZR extract administered at different phases on angiogenesis were investigated to determine the involvement of the upregulation of potential targets through network pharmacology analysis.

Materials And Methods

Liquid chromatography-tandem mass spectrometry profiling combined with network pharmacology predicted therapeutic targets and pathways. Male Sprague-Dawley rats received intracerebroventricular dimethyl sulfoxide 25 min before ischemic stroke induction, followed by intragastric YZR extract (0.8 g/kg) administered during the early subacute phase (D+E-YZR), delayed subacute phase (D+D-YZR), or both the early and delayed subacute phases (D+T-YZR).

Results

Through network pharmacology-based prediction, hypoxia-inducible factor-1α (HIF-1α), vascular endothelial growth factor-A (VEGF-A), and matrix metalloproteinase-9 (MMP-9) signaling pathways are potential targets of YZR extract on angiogenesis in subacute ischemic stroke. D+T-YZR treatment ameliorated neurological deficits; significantly upregulated the expression of phospho-p38 mitogen-activated protein kinase (p-p38 MAPK)/p38 MAPK (1.9-fold), HIF-1α (1.8-fold), nuclear factor E2-related factor 2 (Nrf2) (2.5-fold), VEGF-A (2.3-fold), VEGF receptor 2 (2.7-fold), von Willebrand factor (1.9-fold), stromal cell derived factor-1 (SDF-1) (2.1-fold), C-X-C chemokine receptor type 4 (2.2-fold), angiopoietin-2 (Ang-2) (2.1-fold), Tie2 (2.6-fold), CD105 (1.7-fold), fibroblast growth factor 2 (FGF-2) (2.8-fold), MMP-9 (2.2-fold), nerve growth factor (NGF) (2.4-fold), synapsin 1 (5.4-fold), microtubule-associated protein 2 (3.3-fold), postsynaptic density 95 (1.7-fold), neuronal nuclei (NeuN) (2.3-fold), growth-associated protein 43 (1.8-fold), VEGF-A/CD105 (4.2-fold), and VEGF-A/NeuN (2.5-fold); and downregulated Ang-1 (0.5-fold) expression in the cortical ischemic boundary zone (IBZ). Treatment with the HIF-1α inhibitor 2-methoxyestradiol and the Nrf2 inhibitor ML385 tended to attenuate the effects of D+T-YZR treatment on angiogenesis and neuronal preservation.

Conclusion

YZR extract appeared to promote neurological recovery during both the early and delayed subacute phases of ischemic stroke. This treatment promoted angiogenesis and preserved neurons, with the possible involvement of the upregulation of p38 MAPK-associated HIF-1α/Nrf2/VEGF-A-mediated SDF-1/Ang-2/CD105 and FGF-2/MMP-9/NGF signaling in the cortical IBZ at 28 days post-stroke.