Trans-sodium crocetinate suppresses apoptotic and oxidative response following myoglobin-induced cytotoxicity in HEK-2 cells

Abstract

Purpose: Rhabdomyolysis (RM) is a serious fatal syndrome. The RM leads to acute kidney injury (AKI) as a fatal complication. The belief is that the RM-induced AKI is triggered by myoglobin (MB). MB activates oxidative and apoptotic pathways.Trans-sodium crocetinate (TSC) isobtained from saffron and it has antioxidant andrenoprotective effects. This research was designed to assess the mechanisms of MB-induced cytotoxicity in HEK-2 cells (human embryonic kidney cells) as well as the possible effects of TSC against MB-induced cytotoxicity. Methods: HEK-2 cells were exposed to diverse concentrations of TSC (2.5, 5, 10, 20, 40, 80, and 100 µM) for 24 h. Then, MB (9 mg/ml) was added to the cells. After 24 h, cell viability was measured through MTT, and The values of ROS generation were calculated using DCFH-DA assay. Besides, using western blot autophagy and apoptosis markers in cells were assessed. Results: MB decreased viability and increased ROS levels in HEK-2 cells. Furthermore, MB enhanced the apoptosis markers (cleaved caspase-3 and Bax/Bcl-2 ratio) and autophagy markers (LC3II/I ratio and Beclin-1) in HEK-2 cells. However, pretreatment of HEK-2 cells with TSC for 24 h reduced the cytotoxicity and ROS production caused by MB. Additionally, TSC pretreatment condensed the levels of autophagy and apoptosis markers in response to MB cytotoxicity. Conclusion:TSC has a positive effect in preventing MB-induced cytotoxicity in HEK-2 cells by increasing antioxidant activity and regulation of apoptotic and autophagy signaling pathways.