Astragaloside IV ameliorates high glucose-induced HK-2 cell apoptosis and oxidative stress by regulating the Nrf2/ARE signaling pathway.

Abstract

Diabetic nephropathy (DN) has become the major cause of end-stage renal disease increasing the mortality risk of diabetes. Research has demonstrated that the oxidative damage and apoptosis of renal tubular cells is present during DN. Astragaloside IV (AS-IV) has been widely used for the treatment of many diseases, however, the role and mechanism by which AS-IV may ameliorate high glucose-induced apoptosis and oxidative stress of the human proximal tubular cell line HK-2 remains largely unknown. The present study investigated the effect of AS-IV on high glucose-induced apoptosis and oxidative stress in HK-2 cells. Cell viability, apoptosis and protein expression were detected by Trypan blue staining, Cell Counting Kit-8 assay, terminal deoxynucleotidyl transferase 2′-deoxyuridine-5′-triphosphate nick-end labelling, flow cytometry and western blot analyses. In addition, enzymatic activities, including superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT) and lipid peroxide (LPO), were measured with the corresponding detection kits. DCFH-DA assay and flow cytometry were performed to detect the production of reactive oxygen species (ROS). Western blot analysis and reverse transcription-quantitative polymerase chain reaction were conducted to evaluate protein and mRNA expressions of the nuclear factor erythroid 2 like 2 (Nrf2)/antioxidant response element (ARE) signaling pathway. The results demonstrated that AS-IV significantly enhanced HK-2 cell viability induced by high glucose in a dose-dependent manner. In addition, AS-IV notably inhibited HK-2 cell apoptosis stimulated by high glucose, which may be associated with inhibition of BCL2 associated X protein, Cleaved-caspase-3 and Cleaved-caspase-9, expression and the promotion of Bcl-2. AS-IV significantly increased the activities of antioxidant enzymes SOD, GSH-Px and CAT, and decreased the high-glucose-induced ROS production in HK-2 cells, in a dose-dependent manner. Finally, it was determined that AS-IV regulated the Nrf2/ARE signaling pathway and inhibited the expression of liver-type fatty acid binding protein. In conclusion, these findings may provide evidence that AS-IV has a potential role for the treatment of DN.