β-Asarone Inhibits Amyloid-β by Promoting Autophagy in a Cell Model of Alzheimer's Disease

Abstract

Alzheimer's disease (AD) is one of the most common types of dementia that causes memory, thinking, and behavior problems. The most important feature of AD is the gradual irreversible loss of cognitive ability through the formation of amyloid b (Ab) plaques and neurofibrillary tangles composed of tau protein. The metabolism of Ab and tau proteins is closely related to and is affected by autophagy. Current research speculates that autophagy dysfunction leads to an increase in harmful proteins in AD. b-Asarone is the main constituent of Acorus tatarinowii Schott and has important effects on the central nervous system. In this paper, we primarily explored the effects of b-asarone on the clearance of noxious proteins and the associated potential mechanisms via autophagy in a PC12 cell AD model. A CCK-8 assay and LDH experiments were used to assess cell viability/toxicity, and SPiDER-bGal was used to detect cellular senescence. The important proteins associated with the pathogenesis of AD including APP, PS1, Ab, BACE1, and SYN1 were analyzed by immunofluorescence (IF) and Western blot analysis. Antimycin A (A3) and cyclosporine A (CSA) were selected as the activators and inhibitors of autophagy, respectively. LC3, BECN, P62, PINK1, and Parkin protein expression were also examined by IF and Western blot analysis. The data showed that b-asarone administration significantly dose-dependently increased cell proliferation and decreased cytotoxicity; moreover, b-asarone inhibited SA-bGal and improved cell senescence. The results further showed that, compared to the model, APP, PS1, Ab, BACE1, and p62 were reduced, while SYN1, BECN1, and LC3 were increased after treatment with b-asarone. The results of Canonical Correlation Analysis (CCA) showed a highly significant relationship between the pathological factors of AD and the protein expression of autophagy. In conclusion, our study demonstrated that b-asarone can inhibit Ab, and this effect may occur by promoting autophagy in a cell model of AD.

Sodium Butyrate Protects N2a Cells against Aβ Toxicity In Vitro

“…Aβ is produced by the cleavage of APP by βand γ-secretase [29]. Previous studies have found that the overexpression of APP led to the increase of Aβ, which is closely related to the occurrence of AD [30][31][32][33][34][35][36][37][38][39][40][41][42][43]. It has been shown that BHBA can improve cognitive behavior in an Alzheimer's disease mouse model.…”

“…Cells. Aβ [25][26][27][28][29][30][31][32][33][34][35] (synthesized by Shanghai Sangon Biological Engineering Technology & Services Co.) was diluted in distilled water at a concentration of 16 mmol/L and was maintained at 37°C for 7 days to preage the peptide. N2a cells were divided into four groups: the control (NT) group, the NaB (2 mM) group, the Aβ 25-35 (40 μmol/L) treatment group, and the Aβ 25-35 (40 μmol/L) with NaB (2 mM) pretreatment group.…”

“…N2a cells were divided into four groups: the control (NT) group, the NaB (2 mM) group, the Aβ 25-35 (40 μmol/L) treatment group, and the Aβ 25-35 (40 μmol/L) with NaB (2 mM) pretreatment group. Cells were starved for 4 h and then pretreated with 2 mM NaB for 2 h before Aβ [25][26][27][28][29][30][31][32][33][34][35] was added. RNA or protein from N2a cells was extracted after 24 h. Each experiment was repeated three times.…”

“…After 24 hours, the complete medium was removed, and 200 μL prewarmed DMEM basal medium was added for cell starvation. Subsequently, NaB was added after 4 h to pretreat the cells, and after 2 h, Aβ [25][26][27][28][29][30][31][32][33][34][35] was added for stimulation. There were 5 replicates for each test group.…”

“…Cells were incubated with 0, 40, 60, or 80 μM Aβ 25-35 for 24 hours (n = 4, means ± SD, Student's t-test, * * P < 0:01, and * * * P < 0:001). (b) Effect of 2 mM NaB pretreatment on cell viability challenged by 40 μM Aβ 25-35 for 24 hours.Cells were starved for 4 h and then pretreated with 2 mM NaB for 2 h before Aβ[25][26][27][28][29][30][31][32][33][34][35] was added. Cell viability was analyzed by the CCK-8 assay (n = 4, means ± SD, Student's t-test, * * P < 0:01, and * * * P < 0:001).…”