SAT-185 The role of SIRT2/Mtch1 in vascular calcification of end-stage renal disease involved in ferroptosis

Abstract

Vascular calcification (VC) is a common complication of end-stage renal disease (ESRD) and an important risk factor for the increase of cardiovascular events and mortality in patients with end-stage renal disease. At present, the mechanism of vascular calcification in ESRD patients has not been fully elucidated. The electrolyte imbalances, inflammation, oxidative stress, apoptosis, autophagy and so on are involved in vascular calcification. Among them, Oxidative stress plays an important role in diseases, and lipid peroxides produced in this process are closely related to ferroptosis. Therefore, this study is to explore the further mechanism of VC in ESRD patients. We used vascular smooth muscle cells to established calcification model pretreated with ferroptosis inducer Erastin(Era) and inhibitor Ferrostatin-1(Fer-1). Cells were separated to six groups: Ctrl, Fer, Era, CAL, CAL+Fer and CAL+Era. The cell samples were detedted by Label-free. Ferroptosis was observed by electron microscopy. Mitochondria were stained by mitotracker and its damage was observed by confocal. the production of lipid peroxides including MDA, SOD and NO in cells was detected. Proteins and mRNA expression were detected by western blot and RT-PCR, respectively. In calcified cell model, the level of RUNX2 and OPN were increased. Mitochondrial damage observed in confocal, ferroptosis observed under electron microscopy, MDA and NO production increased, while SOD production decreased. Erastin aggravated but Ferrostatin-1 alleviated calcification. On this basis, we screened out the protein Sirtuin2 (SIRT2) and Mtch1 related to ferroptosis and calcification by Label-free. Western blot demonstrated SIRT2 expression decreased and Mtch1 expression increased in calcified cell model. Fer-1 increased the expression of SIRT2 and thereby decreased Mtch1, while the pretreatment of Erastin inhibited the expression of SIRT2 increased and promoted Mtch1 expression. SIRT2 was involved in vascular calcification of ESRD related to ferroptosis, and its mechanism was regulating the expression of the apoptotic protein Mtch1.