The Combined Therapy of Berberine Treatment with lncRNA BACE1-AS Depletion Attenuates Aβ25-35 Induced Neuronal Injury Through Regulating the Expression of miR-132-3p in Neuronal Cells

Accumulating articles reported that berberine (Ber) played a neuroprotective role in Alzheimer's disease (AD). Long noncoding RNAs (lncRNAs) have been identified as biomarkers and therapeutic targets of AD. However, the precise mechanism by which lncRNA beta-amyloid cleaving enzyme 1 antisense RNA (BACE1-AS)regulates the progression of AD remains largely unknown. HPN and SK-N-SH cells treated with amyloid beta 25-35 (A beta(25-35)) were regarded as AD model in vitro. Cell survival rate was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Lactate dehydrogenase (LDH) cytotoxicity assay was conducted to detect the cytotoxicity of neuronal cells. Flow cytometry was performed to determine the intracellular concentration of Ca2+, reactive oxygen species (ROS) and apoptosis of neuronal cells. Western blot assay was carried out to detect the apoptosis-related proteins of neuronal cells. The abundance of lncRNA BACE1-AS and miR-132-3p was measured by quantitative real time polymerase chain reaction (qRT-PCR). The binding sites between miR-132-3p and BACE1-AS were predicted by Starbase, and the combination was confirmed by dual-luciferase reporter assay. We found that Ber alleviated A beta(25-35) induced neuronal injury in AD model, especially in high concentration Ber group. The enrichment of BACE1-AS was positively regulated by A beta(25-35) and was inversely modulated by Ber in neuronal cells. The interference of BACE1-AS alleviated the neuronal damage of AD model. miR-132-3p was a direct target of lncRNA BACE1-AS in HEK293T cells, and it was negatively regulated by BACE1-AS in neuronal cells. BACE1-AS accumulation reversed the protective effect of miR-132-3p overexpression on AD model. Ber treatment and BACE1-AS intervention recovered the viability of AD model. Ber up-regulated the level of miR-132-3p via BACE1-AS in SK-N-SH and HPN neuronal cells. in conclucsion, Ber protected neuronal cells against A beta(25-35) at least partly through BACE1-AS/miR-132-3p axis. The combined therapy of Ber treatment with BACE1-AS depletion might provide new insight into AD treatment.