Hua Feng Dan protective effect on ischemic stroke through AKT and ERK pathway

Hua Feng Dan (HFD) has demonstrated definitive efficacy in the treatment of ischemic stroke (IS); however, its active components and underlying mechanisms of action remain unclear. This study employed a rat model of middle cerebral artery occlusion (MACO) to evaluate the neuroprotective effects of HFD against cerebral ischemia. Metabolomics approaches were used to demonstrate the complexity of HFD for the treatment of IS. Liquid chromatography/mass spectrometry (LC/MS) was used to identify the main chemical constituents of HFD. Network pharmacology and molecular docking were employed to predict the targets and mechanisms of HFD for IS. Experimental validation was performed to elucidate the underlying mechanisms. HFD demonstrated the ability to enhance neurological function, decrease the area of cerebral infarction, and improve brain structure in rats subjected to MCAO. Twenty-nine distinct metabolites were identified using non-targeted metabolomics analysis of urine. It was determined that HFD mediated its therapeutic effects on IS via amino sugar, nucleotide sugar metabolism, and glycerophospholipid metabolism. Additionally, a total of fifty-six compounds were identified from the alcoholic extracts of HFD, mainly alkaloids, bile acids, flavonoids, and coumarins. The results of network pharmacology and molecular docking suggest that AKT1, STAT3, EGFR, ERK2, and ERK1 are key therapeutic targets. Additionally, HFD upregulates the levels of AKT1 while downregulating those of ERK1/2. This may represent an intrinsic mechanism underlying its anti-cerebral ischemia effects. This study preliminarily elucidates the mechanism of action of HFD in the treatment of IS, providing guidance for its clinical application in this context.