Objective: To investigate the mechanism of miR-223 on the apoptosis of Aβ1-42-induced hippocampal neurons. Methods: The hippocampal neurons of newborn SD rats were cultured in vitro, and randomly divided into blank group(without any transfection), negative group(transfected with miR-NC), miR-223-m group(transfected with miR-223 mimics), miR-223-m+LV group(transfected with miR-223 mimics+lentiviral skeleton), miR-223-m+LV-RhoB group(transfected with miR-223 mimics+Rhob overexpression vector). The expression of miR-223 and RhoB in Aβ1-42-induced hippocampal neurons was detected by qRT-PCR and Western blotting, the survival rate of Aβ1-42-induced hippocampal neurons with different concentrations was detected by MTT assay, and the apoptosis of hippocampal neurons was detected by flow cytometry assay. Dual-luciferase reporter gene was used to detect the fluorescence activity of cells. Results: Aβ1-42 could significantly inhibit the survival rate of hippocampal neurons in a dose-dependent manner, and it also decreased the relative expression of miR-223 and increased the expression level of RhoB. 30 μmol·L-1Aβ1-42 oligopeptide fragment was selected for further grouping. After incubated for 48 hours with 30 μmol·L-1Aβ1-42, the apoptosis rate of the miR-223-m group was lower than that of blank group and negative group, and the apoptosis rate of the miR-223-m+LV-RhoB group was increased compared with the miR-223-m group and the miR-223-m+LV group(P<0.05). In addition, the expression of RhoB protein, Bax and caspase-3 protein was decreased and the expression of Bcl-2 protein was increased in the miR-223-m group compared with the blank group and the negative group(P<0.05), while the expression of Rhob, Bax and caspase-3 protein increased and the expression of Bcl-2 protein decreased in the miR-223-m+LV-RhoB group compared with the miR-223-m group and the miR-223-m+LV group(P<0.05). Dual-Luciferase reporter gene analysis showed that the miR-223 mimics and wild type RhoB 3'UTR could inhibit luciferase activity(P<0.05). Conclusion: miR-223 can inhibit apoptosis of Aβ1-42-induced hippocampal neurons by targeting RhoB. |
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