Effect of trans-sodium crocetinate on contrast-induced cytotoxicity in HEK-۲۹۳ cells

Objective(s): Contrast media (CM) are used for diagnostic or therapeutic intervention purposes in medicine. The main adverse reaction after the administration of CM is contrast-induced nephropathy (CIN). This complication is the third cause of renal failure after hospital treatment. The current study is designed to investigate the possible protective effect of trans-sodium crocetinate (TSC), derived from carotenoid crocetin, against sodium amidotrizoate/meglumine amidotrizoate (SAMA) induced cytotoxicity in HEK-۲۹۳ cells.Materials and Methods: HEK-۲۹۳ cells were incubated with different concentrations of TSC (۱, ۲.۵, ۵, ۱۰, ۲۵, and ۵۰ µM, for ۴۸ hr) and then SAMA (۷ mgI/ml, for ۲۴ hr) was added. The cell viability, intracellular ROS, and phosphatidyl serine exposure were detected by MTT assay, DCFH-DA, and annexin V-FITC/PI method, respectively. The P-ERK/ERK ratio, apoptosis (Bax/Bcl-۲ ratio and cleaved caspase-۳), and autophagy (LC۳ II/I ratio and beclin-۱) markers in cells were evaluated by the western blot method.Results: The exposure of HEK-۲۹۳ cells to SAMA reduced viability, increased apoptotic cells, enhanced ROS production, and subsequently decreased P-ERK/ERK ratio. Similarly, SAMA enhanced apoptosis (Bax/Bcl-۲ ratio and cleaved caspase-۳) and autophagy (LC۳ II/I ratio and beclin-۱) markers in HEK-۲۹۳ cells. The pretreatment of cells with TSC before exposure to SAMA significantly attenuated contrast-induced cytotoxicity. TSC reduced intracellular ROS production and activated the phosphorylation of ERK. In addition, TSC decreased the levels of apoptosis and autophagy proteins. Conclusion: The pretreatment of HEK-۲۹۳ cells with TSC can decrease contrast-induced cytotoxicity through antioxidant effect and modulate  ERK, apoptosis, and autophagy pathways.

Keywords:

Apoptosis , Autophagy , Contrast media , Cytotoxicity , Reactive Oxygen Species , Trans-sodium crocetinate