Qingxin Kaiqiao Fang decreases Tau hyperphosphorylation in Alzheimer's disease via the PI3K/Akt/GSK3β pathway in vitro and in vivo

Ethnopharmacological relevance: Alzheimer's disease (AD) belongs to the category of "senile dementia" in traditional Chinese medicine. AD is associated with brain emptiness or collaterals blocked by phlegm-heat. "Fumanjian" from Jingyue Quanshu treats dementia by promoting qi circulation, alleviating depression, eliminating turbidity, cultivating positivity, and dispelling evil spirits. Qingxin Kaiqiao Fang (QKF), derived from Fumanjian, is effective in treating AD owing to previously mentioned clinical effects. Elucidating the mechanism (s) of action of QKF on AD associated with phlegm-heat may be beneficial for therapeutic management; however, further research is needed. Aim of the study: This study aimed to determine the role of the PI3K/Akt pathway in AD, especially the specific effector protein involved, and explore the efficacy of QKF in treating AD by modulating the PI3K/Akt signal. Materials and methods: High-performance liquid chromatography-Q-orbitrap-mass spectrometry was used to analyze the chemical components of QKF. Subsequently, APP/PS1 double-transgenic mice were used for behavioral tests, and hematoxylin-eosin and Nissl staining were used to assess the neuroprotective and cognitive effects of QKF. Cerebrospinal fluid pharmacology was used in in vitro validation, and A beta 25- 35 was used to induce PC12 cells to establish the AD cell model. Various methods, including immunohistochemistry, Western blotting, quantitative real-time polymerase chain reaction, morphological assay, cell counting kit-8(CCK-8) assay, and terminal deoxynucleotide transferase (TdT)-mediated dUTP nick-end labeling (TUNEL)staining, were used to evaluate the effect of QKF on Tau hyperphosphorylation and anti-apoptosis. These methods also assessed the influence of QKF on the PI3K/Akt/GSK3 beta pathway involving the mRNA and protein expressions. Finally, the inhibitor - LY294002 was used for reverse validation. Results: We identified 295 chemical components in the water extract of QKF.QKF improved spatial cognition and learning memory in APP/PS1 mice, protected PC12 cell morphology, improved cell survival, reduced A beta 25-35induced apoptosis, and inhibited the hyperphosphorylation of Tau protein via the PI3k/Akt/GSK3 beta signaling pathway. Furthermore, this protective effect of QKF was reduced by LY294002 in vitro. Conclusions: QKF can improve spatial cognition, learning, and memory abilities in APP/PS1 mice and protect PC12 cells. Decreasing the Tau hyperphosphorylation in AD exhibits curative efficacy on AD via the PI3K/Akt/ GSK3 beta pathway in vitro and in vivo.