doi: 10.3967/bes2019.039
Effects of Carpobrotus edulis Extract on Oxidative Stress and 158N Oligodendrocyte Death
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Abstract:
Objective Age-related diseases, including neurodegenerative diseases, are associated with oxidative stress and lipid peroxidation, and increase the levels of cholesterol auto-oxidation products such as 7β-hydroxycholesterol (7β-OHC). Thus, it is imperative to identify agents that can prevent 7β-OHC-induced side-effects. Methods We evaluated the potential protective effects of Carpobrotus edulis ethanol-water extract (EWe) on murine oligodendrocytes (158N) cultured in the absence or presence of 7β-OHC (20 μg/mL, 24 h). The cells were incubated with EWe (20-200 μg/mL) 2 h before 7β-OHC treatment. Mitochondrial activity and cell growth were evaluated with the MTT assay. Photometric methods were used to analyze antioxidant enzyme[catalase (CAT) and glutathione peroxidase (GPx)] activities and the generation of lipid and protein oxidation products[malondialdehyde (MDA), conjugated diene (CD), and carbonylated proteins (CPs)]. Results Treatment with 7β-OHC induced cell death and oxidative stress (reflected by alteration in CAT and SOD activities). Overproduction of lipid peroxidation products (MDA and CDs) and CPs was also reported. The cytotoxic effects associated with 7β-OHC were attenuated by 160 μg/mL of EWe of C. edulis. Cell death induced by 7β-OHC treatment was ameliorated, GPx and CAT activities were restored to normal, and MDA, CD, and CP levels were reduced following C. edulis extract treatment. Conclusion These data demonstrate the protective activities of C. edulis EWe against 7β-OHC-induced disequilibrium in the redox status of 158N cells, indicative of the potential role of this plant extract in the prevention of neurodegenerative diseases. -
Figure 1. Effect of Carpobrotus edulis (C. edulis) EWe and 7β-OHC on 158N cell viability. (A) Cells were incubated in the absence or presence of 7β-OHC for 24 h with or without pretreatment with EWe of C. edulis (20-200 µg/mL; 2 h). (B) Cells were incubated in the absence or presence of 7β-OHC for 24 h with or without pretreatment with EWe of C. edulis (140 and 160 µg/mL) for the indicated times. Data shown are mean ± SD from two independent experiments conducted in triplicates. Significance of the difference between vehicle- (ethanol) and EWe- or 7β-OHC-treated cells is indicated with * (Mann- Whitney test, P < 0.05). Significance of the difference between 7β-OHC and 7β-OHC + EWe groups is indicated with # (Mann-Whitney test, P < 0.05).
Figure 2. Effect of Carpobrotus edulis EWe and 7β-OHC on catalase and glutathione peroxidase activities. Cells were incubated in the absence or presence of 7β-OHC for 24 h with or without pretreatment with EWe of C. edulis (160 µg/mL; 6 h). Data shown are mean ± SD from two independent experiments conducted in triplicates. Significance of the difference between vehicle- (ethanol) and 7β-OHC-treated cells is indicated with *. Significance of the difference between 7β-OHC and 7β-OHC + EWe groups is indicated with # (Mann-Whitney test; P < 0.05).
Figure 3. Effect of Carpobrotus edulis EWe and 7β-OHC on lipid peroxidation products. Cells were incubated in the absence or presence of 7β-OHC for 24 h with or without pretreatment with EWe of C. edulis (160 µg/mL; 6 h). Data shown are mean ± SD from two independent experiments conducted in triplicates. Significance of the difference between vehicle- (ethanol) and 7β-OHC-treated cells is indicated with *. Significance of the difference between 7β-OHC and 7β-OHC + EWe groups is indicated with # (Mann-Whitney test; P < 0.05).
Figure 4. Effect of Carpobrotus edulis EWe and 7β-OHC on carbonylated protein levels. Cells were incubated in the absence or presence of 7β-OHC for 24 h with or without pretreatment with EWe of C. edulis (160 µg/mL; 6 h). Data shown are mean ± SD from two independent experiments conducted in triplicates. Significance of the difference between vehicle- (ethanol) and 7β-OHC-treated cells is indicated by *. Significance of the difference between 7β-OHC and 7β-OHC + EWe groups is indicated with # (Mann-Whitney test; P < 0.05).
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