Objective: To investigate the role of miR-424 in diabetic nephropathy (DN) and its relationship with Rictor in mammalian target of rapamycin (mTOR) C2/Akt signaling.
Methods: The western blot analysis and real-time polymerase chain reaction were used to determine the differential expression of Rictor, mTOR, and miR-424 in DN rats. The upregulation of miR-424 was achieved by caudal vein injection of miR-424 mimics. The renal lesion was evaluated by hematoxylin-eosin staining (H&E) and periodic acid schiff staining. The dual-luciferase reporter assay was conducted to determine the binding target of miR-424. The effect of miR-424 upregulation on apoptosis was detected by the terminal deoxynucleotidyl transferase-mediated 2-Deoxyuridine-5-Triphosphate (dUTP) nick-end labeling assay and western blot analysis.
Results: A significantly lower expression of miR-424 and a significantly higher expression of Rictor and mTOR were found in renal tissues of DN rats. The upregulation of miR-424 improved renal lesion and DN symptoms of blood glucose level, urine protein level, body weight, creatinine level, blood urea nitrogen, and KW/BW ratio. The upregulation of miR-424 could significantly reduce apoptosis rates of tissue cells by decreasing the expression levels of caspase-3 and Bax as well as increasing the level of Bcl-2. Furthermore, Rictor was the direct target for miR-424, and upregulation of miR-424 inhibited Rictor through Akt signaling in renal tissue of DN rats and high-glucose-treated human glomerular mesangial cells.
Conclusion: miR-424 contributes to alleviating the symptoms in DN rat models by targeting Rictor through mTORC2/Akt signaling.
Keywords: Akt; Rictor; diabetic nephropathy (DN); mammalian target of rapamycin (mTOR); microRNA-424 (miRNA-424).
© 2019 Wiley Periodicals, Inc.