Neuroprotective Mechanism of Polygonatum sibiricum Polysaccharides in Alzheimer's Disease: Highlighting Role of PI3K-AKT Signalling Pathway and Leptin Receptor

Objective: To investigate the action mechanism of Polygonatum sibiricum Polysaccharides (PSP) in Alzheimer's disease (AD).

Methods: Network pharmacology and molecular docking was used to identify the major active ingredients and potential targets of PSP in treating AD. Male Kunming mice were randomly divided into 6 groups by a simple randomization method: control, model, low-, medium-, and high-dose PSP, and donepezil groups (n=6 per group). An AD mice model was established by intraperitoneally injecting 120 mg/kg D-galactose and oral administration of 40 mg/kg AlCl₃ for 70 d. PSP (100, 200, and 400 mg/kg) and donepezil (5 mg/kg) was administered orally for 35 d, respectively. Behavioral tests including the open field test, elevated plus maze, Morris water maze, and shuttle box test were performed to evaluate anxiety levels and learning and memory abilities. Western blot analysis was used to detect the phosphatidylinositol 3-kinase-protein kinase B (PI3K-AKT) signalling pathway activation. Leptin receptor (LepR) and neuronal nuclei (NeuN) co-localization was observed by immunofluorescence. Adeno-associated virus serotype 9 (AAV9)-mediated LepR knockdown (LepR-KD) was used to investigate the role of LepR in PSP-mediated cognitive improvement in AD mice and LepR and NeuN co-localization in the cerebral cortex. Immunohistochemistry was used to assess Tau Protein deposition in the cortices of AD mice. Enzyme-linked immunosorbent assay quantified pro-inflammatory cytokines levels in the brain tissue.

Results: Network pharmacology identified that PI3K-AKT was the key signalling pathway affected by PSP in AD mice. In vivo experiments showed that PSP significantly improved anxiety levels and cognitive learning abilities in AD mice, upregulated the expression ratios of p-PI3K/PI3K and p-AKT/Akt in brain tissue, enhanced the activity of LepR and NeuN, and reduced Tau Protein accumulation and the expression levels of interleukin (IL)-1β, IL-6, and tumor necrosis factor alpha (P<0.05 or P<0.01). LepR-KD further demonstrated that its deficiency attenuated PSP's neuroprotective effects on cognitive function and cortical neuronal survival.

Conclusion: PSP modulates the PI3K-AKT signalling pathway in a LepR-dependent manner, thereby attenuating aberrant Tau Protein deposition and inflammatory cytokine activity, which may cause delayed AD pathogenesis.

Polygonatum sibiricum polysaccharides; Alzheimer’s disease; Chinese medicine; PI3K-AKT; leptin receptor; network pharmacology.