Institute of Basic Medical Sciences and Peking Union Medical College Hospital, Chinese Academy of Medical Sciences / Peking Union Medical College., 2020.
Objective To explore the role and molecular mechanism of Neferine (Nef) on PC12 cells injury induced by Aβ1-42. Methods PC12 cells were divided into control group, model group(4 μg/mL Aβ1-42 cultured for 24 h) and low, medium and high-dose neferine intervention group. The PC12 cell viability was determined by MTT method and cell apoptosis was detected by flow cytometry. Protein expression of aquaporin4 (AQP4), Bcl-2 and Bax was analyzed by Western Blot, and miR-29a-3p, AQP4 mRNA were detected by qPCR. Biological information prediction and dual luciferase gene report analyzed the targeting relationship between miR-29a-3p and AQP4. miR-29a-3p, si-AQP4 were transfected into PC12 cells induced by Aβ1-42, or anti-miR-29a-3p was transfected into cells and performed high-dose Nef intervention to investigate the effects on cell survival and apoptosisin PC12 cells injury induced by Aβ1-42. Results Compared with the model group, the cell viability rate, Bcl-2 and miR-29a-3p expression of neferine group were significantly increased, and the cell apoptosis rate, Bax, AQP4 mRNA and protein of neferine group expression were significantly decreased (P＜0.05). AQP4 was the target gene of miR-29a-3p. Both over-expression of miR-29a-3p and inhibition of AQP4 expression significantly increased the viability rate of PC12 cells injury induced by Aβ1-42 and the expression level of Bcl-2, and decreased the apoptosis rate and Bax protein level. Inhibitiion of expression of miR-29a-3p reversed the promotion effect of Neferine on the survival of PC12 cell injury induced by Aβ1-42 and the level of Bcl-2 protein, as well as the inhibition effect of Neferine on the apoptosis rate and Bax protein expression. Conclusions Neferine inhibits PC12 cell injury induced by Aβ1-42, promotes cell survival and inhibits apoptosis by miR-29a-3p/AQP4.
