downregulation of mir-205 modulates cell susceptibility to oxidative and endoplasmic reticulum stresses in renal tubular cellsdownregulation mir - 205的调节细胞易受氧化和在肾小管细胞内质网压力.pdfVIP

Downregulation of miR-205 Modulates Cell Susceptibility to Oxidative and Endoplasmic Reticulum Stresses in Renal Tubular Cells Shiyo Muratsu-Ikeda, Masaomi Nangaku*, Yoichiro Ikeda, Tetsuhiro Tanaka, Takehiko Wada, Reiko Inagi* Division of Nephrology and Endocrinology, University of Tokyo School of Medicine, Tokyo, Japan Abstract Background: Oxidative stress and endoplasmic reticulum (ER) stress play a crucial role in tubular damage in both acute kidney injury (AKI) and chronic kidney disease (CKD). While the pathophysiological contribution of microRNAs (miRNA) to renal damage has also been highlighted, the effect of miRNA on renal damage under oxidative and ER stresses conditions remains elusive. Methods: We assessed changes in miRNA expression in the cultured renal tubular cell line HK-2 under hypoxia- reoxygenation-induced oxidative stress or ER stress using miRNA microarray assay and real-time RT-PCR. The pathophysiological effect of miRNA was evaluated by cell survival rate, intracellular reactive oxygen species (ROS) level, and anti-oxidant enzyme expression in miRNA-inhibited HK-2 or miRNA-overexpressed HK-2 under these stress conditions. The target gene of miRNA was identified by 3 9-UTR-luciferase assay. Results: We identified 8 and 10 miRNAs whose expression was significantly altered by oxidative and ER stresses, respectively. Among these, expression of miR-205 was markedly decreased in both stress conditions. Functional analysis revealed that decreased miR-205 led to an increase in cell susceptibility to oxidative and ER stresses, and that this increase was associated with the induction of intracellular ROS and suppression of anti-oxidant enzymes. While increased miR-205 by itself made no change in cell growth or morphol