Data Sheet 1_Astragaloside IV attenuates post-ischemic neuroinflammation associated with Treg-related regulation of CD86/CD206-associated brain myeloid phenotypic responses and PI3K/AKT signaling alterations.zip

Flow cytometry quantified CD4⁺CD25⁺FOXP3⁺ regulatory populations and additionally analyzed CD4⁺FOXP3⁺ populations in the spleen and cervical lymph nodes, together with CD86⁺/CD206⁺ phenotypic responses within the CD11b⁺CD45^low/int population in brain tissue. Immunofluorescence evaluated brain CD3⁺FOXP3⁺ signals and Iba1/CD86 or Iba1/CD206 double-positive signals.

Anti-CD25 intervention was used to evaluate the involvement of Treg-related immune responses in AS-IV-associated effects.</p>Results<p>AS-IV reduced infarct volume, showed a trend toward improvement in neurological deficit scores, and alleviated histopathological injury after MCAO. It decreased IL-6 and TNF-α and increased IL-10 and TGF-β in ischemic brain tissue. Transcriptomic analysis identified changes associated with the PI3K/AKT signaling pathway and immune regulatory processes, while western blotting showed increased p-PI3K/PI3K and p-AKT/AKT ratios after AS-IV treatment.

AS-IV increased Treg proportions in the spleen and cervical lymph nodes and increased CD3⁺FOXP3⁺ signals in brain tissue. It also reduced CD86⁺-associated and increased CD206⁺-associated brain myeloid phenotypic responses. Anti-CD25 partially attenuated AS-IV-associated improvements in neurological and brain injury outcomes, as well as its effects on inflammatory cytokines and brain myeloid phenotypic responses.</p>Conclusion<p>AS-IV attenuated post-ischemic neuroinflammation in association with Treg-related immune responses and PI3K/AKT-associated signaling alterations, accompanied by changes in CD86/CD206-associated brain myeloid phenotypic responses.

These findings support AS-IV as a potential neuroimmunomodulatory intervention for ischemic stroke.</p>