P0526MIR-181D-5P TARGETS KLF6 TO IMPROVE ISCHEMIC/REPERFUSION AKI THROUGH EFFECTS ON RENAL FUNCTION,APOPTOSIS AND INFLAMMATION

Abstract

Abstract Background and Aims Inflammation and renal tubular cell death is the most crucial pathophysiological changes in acute kidney ischemic/reperfusion injury(IRI). The miR-181d family plays diverse roles in cell proliferation, apoptosis and inflammation, but its renal target and potential role in IRI are unknown. Method We built hypoxia/reoxygenation (H/R) renal cell model (HK-2 cells) and renal ischemia/reperfusion (I/R) injury mice model. We used observe the expression location of miR-181d-5p and KLF6 by in situ hybridization and immunohistochemistry, the blood creatinine and urea nitrogen levels through fully automatic biochemical detector, the severity of kidney injury by HE staining, the apoptosis and necrosis of renal tubular epithelial cells by TUNEL and Annexin V-FITC/PI flow cytometry, the concentrations of pro-inflammatory factors in the cell supernatant by ELISA and the mRNA and protein levels of related factors and predicted and verified the relationship between miR-181d-5p and KLF6 via bioinformatics systems and luciferase reporter genes. Results We showed that the expression of miR-181d-5p decreased in HK-2 cells of H/R and renal mouse model of I/R injury. Mechanistically, KLF6 was predicted as a new potential target gene of miR-181d-5p through bioinformatics and the Luciferase reporter verification. After renal ischemia/ reperfusion, overexpression of microRNA-181d significantly attenuated the level of KLF6 and inhibited inflammatory mediators such as IL-6, TNF-α, reducing apoptosis and further improving renal function. Critical NF-κB signaling pathways mediated this inflammatory response. Conclusion In conclusion, upregulation of microRNA-181d inhibits KLF6 expression, which have protective anti-apoptosis and anti-inflammatory effects against I/R kidney in vivo, and H/R HK-2 cells in vitro, thereby providing novel insights into the molecular mechanisms associated with ischemia/reperfusion injury and a potential novel therapeutic target.