[논문]Protective Role of Corticosterone against Hydrogen Peroxide-Induced Neuronal Cell Death in SH-SY5Y Cells

Lee, Chan; Jang, Jung-Hee; Park, Gyu Hwan

doi:10.4062/biomolther.2022.126

Protective Role of Corticosterone against Hydrogen Peroxide-Induced Neuronal Cell Death in SH-SY5Y Cells 원문보기

Biomolecules & therapeutics, v.30 no.6, 2022년, pp.570 - 575

Lee, Chan (Department of Pharmacology, School of Medicine, Keimyung University) , Jang, Jung-Hee (Department of Pharmacology, School of Medicine, Keimyung University) , Park, Gyu Hwan (College of Pharmacy, Research Institute of Pharmaceutical Sciences, Kyungpook National University)

Abstract ▼ AI-Helper

Stress breaks body balance, which can cause diverse physiological disorders and worsen preexisting diseases. However, recent studies have reported that controllable stress and overcoming from stress reinforce resilience to resist against more intense stress afterwards. In this study, we investigated the protective effect of corticosterone (CORT), a representative stress hormone against hydrogen peroxide (H2O2)-induced neuronal cell death and its underlying molecular mechanism in SH-SY5Y cells, a human neuroblastoma cell line. The decreased cell viability by H2O2 was effectively restored by the pretreatment with low concentration of CORT (0.03 μM for 72 h) in the cells. H2O2-increased expression of apoptotic markers such as PUMA and Bim was decreased by CORT pretreatment. Furthermore, pretreatment of CORT attenuated H2O2-mediated oxidative damages by upregulation of antioxidant enzymes via activation of nuclear factor erythroid 2-related factor 2 (Nrf2). These findings suggest that low concentration of CORT with eustressed condition enhances intracellular self-defense against H2O2-mediated oxidative cell death, suggesting a role of low concentration of CORT as one of key molecules for resilience and neuronal cell survival.

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그림 Fig. 1. Effect of CORT on cell viability in SH-SY5Y cells. The cells were treated with various concentration of CORT for 24 h. (A) High concentration groups were treated with 1, 5, 10, 25, 50 and 100 μM CORT. (B) Low concentration groups were treated with 0.003, 0.01, 0.03, 0.01, 0.3 and 1 μM CORT. Cell viability was determined by using MTT assay. Data were presented as mean ± SEM (n=3), **p<0.01 vs. control group.
그림 Fig. 2. Effect of CORT against H₂O₂-decreased cell viability in SHSY5Y cells. The cells were pretreated with 0.003 and 0.03 μM of CORT for 72 h and then incubated with or without 250 μM H₂O₂ for 24 h. Cell viability was determined by using MTT assay. Data were presented as mean ± SEM (n=3), **p<0.01 vs. control group; ^##p<0.01 vs. H₂O₂-treated group.
그림 Fig. 3. Effect of CORT on H₂O₂-induced PUMA expression in SHSY5Y cells. The cells were pretreated with 0.003 and 0.03 μM CORT for 72 h and then incubated with or without 250 μM H₂O₂ for 24 h. PUMA expression was determined by western blot (A), and presented a relative intensity of PUMA/Actin (B). Data were expressed as mean ± SEM (n=3), **p<0.01 vs. control group; ^##p<0.01 vs. H₂O₂-treated group.
그림 Fig. 4. Effect of CORT on H₂O₂-induced Bim expression in SHSY5Y cells. The cells were pretreated with 0.003 and 0.03 μM CORT for 72 h and then incubated with or without 250 μM H₂O₂ for 24 h. Bim expression was determined by western blot (A), and presented a relative intensity of Bim/Actin (B). Data were expressed as mean ± SEM (n=3), **p<0.01 vs. control group; ^##p<0.01 vs. H₂O₂-treated group.
그림 Fig. 5. Effect of CORT against H₂O₂-induced 4-HNE expression and ROS production in SH-SY5Y cells. The cells were pretreated with 0.003 and 0.03 μM CORT for 72 h and then incubated with or without 250 μM H₂O₂ for 24 h. 4-HNE expression was determined by western blot assay (A) and presented a relative intensity of 4-HNE/Actin (B). Intracellular ROS was determined by using DHR 123 fluorescence dye (C). Data were expressed as mean ± SEM (n=3), **p<0.01 vs. control group; ^#p<0.05 and ^##p<0.01 vs. H₂O₂- treated group.
그림 Fig. 6. CORT-induced expression of antioxidant enzymes in SHSY5Y cells. The cells were pretreated with 0.003 and 0.03 μM CORT for 72 h and then incubated with or without 250 μM H₂O₂ for 24 h. Expression levels of HO-1, GCS and MnSOD were measured by western blot assay (A) and presented a relative intensity of HO-1/Actin, GCS/Actin and MnSOD/Actin (B). Data were expressed as mean ± SEM (n=3), ^#p<0.05 and ^##p<0.01 vs. H₂O₂- treated group.
그림 Fig. 7. CORT-recovered activation of Nrf2 in SH-SY5Y cells The cells were pretreated with 0.003 and 0.03 μM CORT for 72 h and then incubated with or without 250 μM H₂O₂ for 24 h. Phosphorylation of p-Nrf2 at Ser40 were determined by western blot assay (A) and calculated a relative intensity of p-Nrf2/Nrf2 (B). Data were presented as mean ± SEM (n=3), ^##p<0.01 vs. H₂O₂-treated group.

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