Abstract
BackgroundmiR-214 was demonstrated to be upregulated in models of renal disease and promoted fibrosis in renal injury independent of TGF-β signaling in vivo. However, the detailed role of miR-214 in acute kidney injury (AKI) and its underlying mechanism are still largely unknown.MethodsIn this study, an I/R-induced rat AKI model and a hypoxia-induced NRK-52E cell model were used to study AKI. The concentrations of kidney injury markers serum creatinine, blood urea nitrogen, and kidney injury molecule-1 were measured. The expressions of miR-214, tumor necrosis factor-α, interleukin (IL)-1β, IL-6, were detected by RT-qPCR. The protein levels of Bcl-2, Bax, Dickkopf-related protein 3, β-catenin, c-myc, and cyclinD1 were determined by western blot. Cell apoptosis and caspase 3 activity were evaluated by flow cytometry analysis and caspase 3 activity assay, respectively. Luciferase reporter assay was used to confirm the interaction between miR-214 and Dkk3.ResultsmiR-214 expression was induced in ischemia–reperfusion (I/R)-induced AKI rat and hypoxic incubation of NRK-52E cells. Overexpression of miR-214 alleviated hypoxia-induced NRK-52E cell apoptosis while inhibition of miR-214 expression exerted the opposite effect. Dkk3 was identified as a target of miR-214. Anti-miR-214 abolished the inhibitory effects of DKK3 knockdown on hypoxia-induced NRK-52E cell apoptosis by inactivation of Wnt/β-catenin signaling. Moreover, miR-214 ameliorated AKI in vivo by inhibiting apoptosis and fibrosis through targeting Dkk3 and activating Wnt/β-catenin pathway.ConclusionmiR-214 ameliorates AKI by inhibiting apoptosis through targeting Dkk3 and activating Wnt/β-catenin signaling pathway, offering the possibility of miR-214 in the therapy of ischemic AKI.
Highlights
MiR-214 was demonstrated to be upregulated in models of renal disease and promoted fibrosis in renal injury independent of TGF-β signaling in vivo
Since inflammation and tubular epithelial cell apoptosis are the major features of acute kidney injury (AKI) [25], the effectiveness of hypoxia-induced NRK-52E cell model was further assessed by detecting the expression of inflammatory factors including tumor
Overexpression of miR‐214 alleviated hypoxia‐induced apoptosis in NRK‐52E cells The expression of miR-214 in kidney tissues of I/Rinduced rat AKI models was analyzed by qRT-PCR and the in vivo results demonstrated that miR-214 expression was aberrantly upregulated in I/R-induced rat AKI models after 12, 48 and 72 h of reperfusion compared to that in sham group (Fig. 2a)
Summary
MiR-214 was demonstrated to be upregulated in models of renal disease and promoted fibrosis in renal injury independent of TGF-β signaling in vivo. The detailed role of miR-214 in acute kidney injury (AKI) and its underlying mechanism are still largely unknown. Previous studies discovered that miR-214 was up-regulated in models of renal disease and promoted fibrosis in renal injury independent of TGF-β signaling in vivo [11, 12]. Dickkopf (DKK) proteins, an evolutionarily conserved family consisting of DKK1, 2, 3, and 4, have been reported to function as regulators of Wnt/β-catenin pathway [18]. They encode secreted proteins that typically antagonize Wnt/β-catenin signaling, by inhibiting the Wnt coreceptors LRP5 and 6 [19]. This study was designed to investigate the functions and mechanisms of miR-214 in I/R-induced AKI rat model and hypoxiainduced NRK-52E cell model
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