Objective:

2

To investigate the action mechanism of

Polygonatum sibiricum

polysaccharides (PSP) in Alzheimer's disease (AD).

Methods:

2

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 AlCl

3

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:

2

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:

2

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.