The Treatment of Thymoquinone Increased Expression Levels of Apoptotic and Oxidative Genes in the Nrk-52e Rat Kidney Epithelial Cell Line

Abstract

This study was aimed to show the effects of Thymoquinone (TQ), the active ingredient of Nigella sativa, on apoptotic and oxidative pathways in kidney cells, depending on concentration and time. For this purpose, the NRK-52E rat kidney epithelial cell line was used. To determine the IC50 and proliferative values of TQ, the MTT cell viability test was performed at 24 and 48 hours. The proliferative (TQP-10µM) and toxic (TQIC50-60 µM) concentrations were determined. The expression of apoptotic (caspase 3 (CASP3), caspase 8 (CASP8), caspase 9 (CASP9), Bax (Bcl-2 associated X), Bcl-2 (Bcl-2 apoptosis regulator)) genes and oxidative (glutathione peroxidase 1(GPX1)), superoxide dismutase 1 (SOD1), neutrophil cytosolic factor 1 (NCF1)) genes were determined as concentration and time-dependent, in the samples taken by RT-qPCR at the concentrations and times dependent. According to the results of this analysis, GPX1 gene was significantly up-regulated in Tp compared to the control gene at 24 hours, SOD1 gene and NCF1 gene did not change in TIC50. Apoptotic genes were found to be limited up-regulated at both concentrations at 24 hours. At 48 hours, GPX1 was up-regulated at TQP concentration and SOD1 at TQIC50 concentration. The NCF1 gene was significantly expressed at TPIC50 at only 48 hours. Apoptotic genes were limitedly upregulated at TQP concentration at 48 hours, while significant upregulation of all apoptotic genes was found at TQIC50 concentration. In conclusion, GPx increased significantly at Tp at the 24th hour, SOD1 at TQIC50 concentration, and NCF gene increased significantly at the toxic concentration at the 48th hour. It was revealed that the effect of the apoptotic pathway on TQ-dependent cell death was limited at the 24th hour, while apoptotic cell death at toxic concentrations occurred externally at the 48th hour. © 2023 Suleyman Demirel University. All rights reserved.