Allicin induces post-translational modifications of p53, DNA damage, and oxidative stress in human embryonic kidney cells

• Published in: Journal of biotech research 2022-01, Vol.13, p.90-102
• Main Authors: Narain, Nicole M, Amoako, Daniel G, Somboro, Anou M, Arhin, Isaiah, Mhlongo, Ndumiso N, Kumalo, Hezekiel M, Khan, Rene B
• Format: Article
• Language: English
• Subjects: Adenosine diphosphate; Adenosine triphosphate; Antioxidants; Apoptosis; ATP; BAX protein; Bcl-2 protein; Bioassays; Caspase; Cell culture; Cell death; Comet assay; Cytotoxicity; Deoxyribonucleic acid; DNA; DNA damage; DNA fragmentation; Enzymes; Fragmentation; Free radicals; Garlic; Gene expression; Kidneys; Lipid peroxidation; Lipids; Nitrogen; Oxidants; Oxidative stress; Oxidizing agents; Oxygen; p53 Protein; Peroxidation; Physiology; Poly(ADP-ribose) polymerase; Post-translation; Proteins; Reactive nitrogen species; Reactive oxygen species; Ribose; Superoxide dismutase; Toxicity; Tumors; Western blotting
• ISSN: 1944-3285

Allicin derived from garlic has been exploited as a therapeutic intervention in the alleviation of fatal diseases. However, its molecular effect on the kidney is yet to be elucidated. This study aimed to determine the toxicity of allicin by assessing the oxidative and apoptotic pathways in human embryonic kidney (HEK293) cells. The HEK293 cells were cultured until confluent in complete culture medium, then treated with the aqueous allicin extract for 24 hours. Cytotoxicity was determined using the 3-(4,5-dimethylthiazol-2-yl)2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. Nitrates and lipid peroxidation assays were performed to assess the generation of free radicals and reactive oxygen/nitrogen species. Luminometry was used to assess caspase activation and adenosine triphosphate concentration. Fragmentation of DNA was measured utilizing the comet assay and confirmed through Hoechst staining. Additionally, protein expressions of nuclear factor erythroid 2-related factor 2 (Nrf2), superoxide dismutase 2 (SOD2), Bcl-2 associated X-protein (Bax), tumor protein p53, and Poly (ADP-ribose) polymerase 1 (PARP-1) were examined by Western blotting assay. Cell viability was decreased with increasing allicin concentration. Although the antioxidant SOD2 and its regulator Nrf2 were upregulated, the higher concentrations of allicin incited its oxidant capacity. This was affirmed by a dose-dependent increase in both reactive nitrogen species and reactive oxygen species that overwhelmed allicin's antioxidant effects and resulted in oxidative stress. Additionally, allicin induced apoptosis via caspase activation, adenosine triphosphate stimulation, cleavage of PARP-1, and up-regulation of Bax. Fragmentation of DNA was confirmed by increased comet tail length and the apoptotic cells displayed in the Hoechst assay. Moreover, allicin treatment downregulated the expression of p53 while upregulating PARP-1. Allicin is cytotoxic to HEK293 cells via oxidative stress and apoptotic cell death.