Renshen Guipi Wan alleviates chronic cerebral hypoperfusion-induced cognitive impairment in rats by modulating PI3K/Akt and Notch1/HES1 signaling

Objective:  Renshen Guipi Wan (RSGPW) is derived from Guipi Tang and has traditionally been used to treat forgetfulness. We characterized its chemical profile and investigated its effects on cognitive impairment associated with chronic cerebral hypoperfusion.   Methods : RSGPW was analyzed by ultra-high performance liquid chromatography coupled with Q Exactive HF Orbitrap high-resolution mass spectrometry (UHPLC-HRMS). Rats subjected to permanent bilateral common carotid artery occlusion (2VO) received RSGPW (1, 2, or 4 g/kg/day by gavage) or donepezil (5 mg/kg/day) for 8 weeks. Cognitive function, cortical perfusion, hippocampal injury, redox markers, and synaptic and signaling proteins were assessed. Network analysis guided the investigation of signaling pathways in H₂O₂-injured SH-SY5Y cells treated with RSGPW-containing serum. LY294002 and valproic acid were used to pharmacologically modulate the respective pathways.   Results : A total of 140 compounds were tentatively identified. RSGPW improved spatial and recognition memory, increased cortical perfusion, reduced hippocampal neuronal injury, increased superoxide dismutase activity, decreased malondialdehyde levels, and restored the expression of postsynaptic density protein 95 and synaptophysin. It also increased hippocampal phosphoinositide 3-kinase (PI3K) expression and protein kinase B (Akt) phosphorylation while reducing Notch1 and hairy and enhancer of split-1 (HES1). Similar changes were observed in H₂O₂-injured cells. LY294002 attenuated the RSGPW-associated increase in Akt phosphorylation, whereas valproic acid increased Notch1 and HES1 in RSGPW-treated cells.   Conclusion : RSGPW alleviated hypoperfusion-associated cognitive impairment, accompanied by improved cortical perfusion and redox balance, recovery of synaptic protein expression, restoration of PI3K/Akt signaling, and suppression of Notch1/HES1 signaling. These findings provide preclinical support for further investigation of RSGPW as a potential treatment for hypoperfusion-related cognitive impairment.