SUN-081 THE EFFECT OF ASTRAGALOSIDE-IV ON OXYGEN STRESS AND EMT OF HMRSV5 INDUCED BY HIGH-GLUCOSE PERITONEAL DIALYSATE

Abstract

Epithelial-to-mesenchymal transition (EMT) of peritoneal mesothelial cells (PMCs) is the crucial process of peritoneal fibrosis (PF), which rise great challenge for peritoneal dialysis therapy. This study aims to observe the effect of Astragaloside-IV (AS-IV) on oxygen stress and EMT of HMrSV5 PMCs, explore the function of ROS in EMT and pharmacological mechanism of AS-IV. We used peritoneal dialysis solutions (PDS) with high glucose concentration to establish oxygen stress model of HMrSV5 cells. Morphologic alterations of cells were observed and MTT assay was used to detect cell viability. DHE fluorescent probes were loaded to label secreted ROS. EMT markers and signal transduction factors were analyzed by western blotting. 40 or 50 μg/mL of AS-IV increased the viability of HMrSV5, while it was suppressed due to the longstanding explosure to PDS with high glucose concentration, notably in PDS containing 4.25% glucose for 48h. Phenotypic changes of HMrSV5 were characterized by cell elongation and losing cobblestone like feature. 40μg/mL of AS-IV could suppress morphologic changes of cells, decrease ROS expression in model group, upregulate the protein expression of E-cadherin and ZO-1 while downregulate α-SMA. Furthermore, the phosphorylation of Smad2/3 protein was inhibited by AS-IV, and 5mmol/L of NAC had the similar effect on EMT of PMCs. PDS with high glucose concentration can suppress cell viability and induce EMT of PMCs by upregulating ROS expression and activation of Smads pathway. AS-IV could reduce ROS generation, suppress the phosphorylation of Smad2/3 and thus, inhibit the EMT of PMCs.