Tetrandrine ameliorated Alzheimer's disease through suppressing microglial inflammatory activation and neurotoxicity in the 5XFAD mouse

Abstract

Background: Alzheimer's disease (AD) is a neurodegenerative disorder prevalent in the aged population. Tetrandrine is a natural metabolite isolated from herbal medicine Stephania tetrandra with various activities.

Purpose: In this study, we investigated the therapeutic role of tetrandrine in 5XFAD mouse, a transgenic model of AD.

Methods: 5XFAD mice were intraperitoneally injected with saline or different doses of tetrandrine (10, 20, and 40 mg/kg per 2 days) from the age of 5 months to 7 months followed by the determination of cognitive ability, amyloid plaque load, cell apoptosis, and inflammation in the brain. In vitro, the protective roles of tetrandrine against inflammatory activation of microglia and the resulting neurotoxicity were studied in BV2 cells and differentiated PC12 cells, respectively.

Results: Morris water maze test showed that two months of tetrandrine treatment dose-dependently improved the cognitive ability of 5XFAD mice. Immunostaining against Aβ 1-42 demonstrated reduced amyloid plaque deposition in the brain of tetrandrine-treated 5XFAD mice. TUNEL assay revealed decreased cell apoptosis in the hippocampus after tetrandrine treatment. Further, RT-PCR showed that the ectopic transcription of inflammation-associated genes including TNFα, IL-1β, IL-6, COX-2, iNOS, and p65 was reversed in 5XFAD mice treated with tetrandrine. In vitro, Aβ 1-42 stimulated the secretion of inflammatory cytokines TNFα and IL-1β in microglial BV2 cells as determined by ELISA, which was suppressed by tetrandrine pre-treatment. Tetrandrine pre-treatment also inhibited the expression of TLR4, p65, iNOS, and COX-2 in BV2 cells induced by Aβ 1-42. Most importantly, treatment of PC12-derived neuron-like cells with conditional medium from Aβ 1-42-stimulated BV2 cells remarkably impaired cell viability and promoted cell apoptosis, which was attenuated by the conditional medium from BV2 cells with tetrandrine pre-treatment.

Conclusion: Collectively, findings in this study demonstrated that tetrandrine ameliorates AD by suppressing microglia-mediated inflammation and neurotoxicity.

Keywords: 5XFAD mouse; Alzheimer's disease; Inflammation; Microglia; Neurotoxicity; Tetrandrine.