Herbal formula FBD extracts prevented brain injury and inflammation induced by cerebral ischemia–reperfusion

The aim of this work was to verify neuroprotective and anti-inflammatory properties of FBD, a herbal formula composed of Poria cocos, Atractylodes macrocephala and Angelica sinensis, in ICR mice subjected to repetitive 10 min of common carotid arteries occlusion followed 24 h reperfusion. Intragastrical pretreatment with supercritical carbon dioxide extract (FBD-CO2, 37.5 mg/kg) twice daily for 3.5 d, significantly reduced Evans Blue influx, neuron specific enolase (NSE) efflux, brain infarction (all p < 0.05), also inhibited polymorphonuclear leukocytes (PMNs) infiltration (p < 0.001), suppressed secretion of tumor necrosis factor (TNF)-α in blood (p < 0.05), interleukin (IL)-1β and IL-8 in brain (both p < 0.01), and down-regulated cerebral expression of phosphor-IκB-α and phosphor-nuclear factor kappa-B (NF-κB), whether coupled with aqueous extract (FBD-H2O, 150 mg/kg) or not. Moreover, FBD-CO2 (0.1–10 μg/ml) inhibited 0.1 μM phorbol myristate acetate-evoked oxidative burst in rat PMNs, 20 ng/ml TNF-α-triggered PMNs adhesion to ECV304 endothelial cells, and PMNs neurotoxicity to PC12 neuron-like cells as well as NSE release (IC50 1.30, 0.98, 0.24 and 0.82 μg/ml, respectively). Our study demonstrated that FBD-CO2 prevented brain ischemia/reperfusion injury, at least in part, by limiting PMNs infiltration and neurotoxicity mediated by TNF-α, IL-1β and IL-8, via inhibition on NF-κB activation.