Forkhead box A2-mediated lncRNA SOX2OT up-regulation alleviates oxidative stress and apoptosis of renal tubular epithelial cells by promoting SIRT1 expression in diabetic nephropathy.

Abstract

Renal tubular injury is the main feature of diabetic nephropathy (DN). We intend to investigate the function and related mechanisms of lncRNA SOX2 overlapping transcript (SOX2OT) in high glucose (HG)-induced oxidative stress and apoptosis of renal tubular epithelial cells (RTECs). To construct diabetes models, the human kidney-2 (HK-2) cells were treated with HG (30 mM), and mice were injected with streptozotocin. The levels of intracellular and mitochondrial reactive oxygen species (ROS) were assessed by dihydroethidium staining and MitoSox staining. The cell apoptosis was assessed by flow cytometry and TUNEL staining. Levels of serum creatinine, blood urea nitrogen (BUN), Urinary ACR, and oxidative stress marker 8-hydroxy-2'-deoxyguanosine (8-OHdG) were detected by relevant kits. In addition, fluorescence in situ hybridization staining, RNA-pull down, RNA immunoprecipitation (RIP), co-immunoprecipitation (co-IP), dual-luciferase reporter gene assay and chromatin immunoprecipitation (ChIP) were also executed. Levels of SOX2OT and silent information regulator 1 (SIRT1) were down-regulated in HG-cultured HK-2 cells. Overexpressing SOX2OT reduced intracellular and mitochondrial ROS levels and cell apoptosis in vitro. Moreover, SOX2OT overexpression also reduced serum creatinine, BUN, urinary ACR, 8-OHdG, renal tubular injury markers KIM1 and NGAL, ROS levels, and cell apoptosis in vivo. In addition, SOX2OT promoted SIRT1 expression by suppressing its ubiquitination. Besides, interference with SIRT1 reversed the inhibitory effect of SOX2OT overexpression on HG-induced oxidative stress and apoptosis. Forkhead box A2 (Foxa2) levels were up-regulated in HG-cultured HK-2 cells. Foxa2 could bind to the SOX2OT promoter and suppress its expression. Furthermore, interfering with SOX2OT reversed the inhibitory effect of Foxa2 interference on HG-induced oxidative stress and apoptosis. Foxa2-mediated SOX2OT up-regulation reduced oxidative stress and apoptosis of RTECs by promoting SIRT1 expression, thus alleviating the progression of DN.