[논문]Protective effect of Capsosiphon fulvescens on oxidative stress-stimulated neurodegenerative dysfunction of PC12 cells and zebrafish larva models

Laxmi Sen Thakuri; Jung Eun Kim; Jin Yeong Choi; Dong Young Rhyu

doi:10.47853/fas.2023.e2

Protective effect of Capsosiphon fulvescens on oxidative stress-stimulated neurodegenerative dysfunction of PC12 cells and zebrafish larva models 원문보기

Fisheries and aquatic sciences, v.26 no.1, 2023년, pp.24 - 34

Laxmi Sen Thakuri (Department of Nutraceutical Resources, Mokpo National University) , Jung Eun Kim (Department of Nutraceutical Resources, Mokpo National University) , Jin Yeong Choi (Department of Nutraceutical Resources, Mokpo National University) , Dong Young Rhyu (Department of Nutraceutical Resources, Mokpo National University)

Abstract ▼ AI-Helper

Reactive oxygen species (ROS) at high concentrations induce oxidative stress, an imbalanced redox state that is a prevalent cause of neurodegenerative disorders. This study aimed to investigate the protective effect of Capsosiphon fulvescens (CF) extract on oxidative stress-induced impairment of cognitive function in models of neurodegenerative diseases. CF was extracted with subcritical water and several solvents and H2O2 (0.25 mM) or aluminum chloride (AlCl3; 25 µM) as an inducer of ROS was treated in PC12 neuronal cells and zebrafish larvae. All statistical analyses were performed using one-way analysis of variance and Dunnett's test using GraphPad Prism. H2O2 and AlCl3 were found to significantly induce ROS production in PC12 neuronal cells and zebrafish larvae. In addition, they strongly affected intracellular Ca2+ levels, antioxidant enzyme activity, brain-derived neurotrophic factor (BDNF) and tropomyosin receptor kinase B (TrkB) signaling, acetylcholinesterase (AChE) activity, and hallmarks of Alzheimer's disease. However, treatment of H2O2-induced PC12 cells or AlCl3-induced zebrafish larvae with CF subcritical water extract at 90℃ and CF water extract effectively regulated excessive ROS production, intracellular Ca2+ levels, and mRNA expression of superoxide dismutase, glutathione peroxide, glycogen synthase kinase-3 beta, β-amyloid, tau, AChE, BDNF, and TrkB. Our study suggested that CF extracts can be a potential source of nutraceuticals that can improve the impairment of cognitive function and synaptic plasticity by regulating ROS generation in neurodegenerative diseases.

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표/그림 (8)

그림 Fig. 1. Effect of CF on viability of PC12 neuronal cells. Data are presented as mean ± SE (n = 12) and analyzed by one-way analysis of variance followed by Dunnett test. *p < 0.05, **p < 0.01, ***p < 0.001 vs. untreated normal cells. CF, Capsosiphon fulvescens; Normal, untreated CF extracts; CF90, CF subcritical water extract at 90℃ and 3 MPa; CF150, CF subcritical water extract at 150℃ and 3 MPa; CF210, CF subcritical water extract at 210℃ and 3 MPa; CFW, CF water extract; CFM, CF methanol extract; CFE, CF ethanol extract.
그림 Fig. 2. Effect of H₂O₂ or CF on viability of PC12 neuronal cells. Data are presented as mean ± SE (n = 12) and analyzed by one-way analysis of variance followed by Dunnett test. **p < 0.01, ***p < 0.001 vs. untreated normal cells; ^###p < 0.001 vs. H₂O₂-treated cells. CF, Capsosiphon fulvescens; Normal, untreated CF extracts; CF90, CF subcritical water extract at 90℃ and 3 MPa pressure; CF150, CF subcritical water extract at 150℃ and 3 MPa pressure; CF210, CF subcritical water extract at 210℃ and 3 MPa pressure; CFW, CF water extract; CFM, CF methanol extract; CFE, CF ethanol extract.
그림 Fig. 3. Effect of CF90 and CFW on reactive oxygen species production (A), acetylcholinesterase enzyme activity (B), and calcium level (C) in H₂O₂-induced PC12 neuronal cells. The concentration of H₂O₂ was 0.25 mM. Data are presented as mean ± SE (n = 6) and analyzed by one-way analysis of variance followed by Dunnett test. *p < 0.05, **p < 0.01, ***p < 0.001 vs. untreated normal cells; ^#p < 0.05, ^##p < 0.01, ^###p < 0.001 vs. H₂O₂- treated cells. CF, Capsosiphon fulvescens; Normal, untreated CF extracts; CF90, CF subcritical water extract at 90℃ and 3 MPa; CFW, CF water extract.
그림 Fig. 4. Effect of AlCl3 on survivability (A) and reactive oxygen species production (B and C) in zebrafish larvae. Magnification was × 20. Data are presented as mean ± SE (n = 8) and analyzed by one-way analysis of variance followed by Dunnett test. **p < 0.01 vs. untreated normal zebrafish larvae. AlCl₃, aluminum chloride.
그림 Fig. 5. Effect of CF90 and CFW extracts on AlCl₃-induced reactive oxygen species production in zebrafish larvae. The concentration of AlCl₃ was 25 μM. Magnification was × 20. Data are presented as mean ± SE (n = 12) and analyzed by one-way analysis of variance followed by Dunnett test. ***p < 0.001 vs. untreated normal zebrafish larvae; ^##p < 0.01, ^###p < 0.001 vs. AlCl₃- treated zebrafish larvae. CF, Capsosiphon fulvescens; Normal, untreated CF extracts; CF90, CF subcritical water extract at 90℃ and 3 MPa pressure; CFW, CF water extract; AlCl₃, aluminum chloride.
그림 Fig. 6. Effect of CF90 and CFW on mRNA expression of antioxidant enzyme in AlCl₃-induced zebrafish larvae. The concentration of AlCl₃ was 25 μM. Data are presented as mean ± SE (n = 3) and analyzed by one-way analysis of variance followed by Dunnett test. *p < 0.05, **p < 0.01 vs. untreated normal zebrafish larvae; #p < 0.05, ^##p < 0.01, ^###p < 0.001 vs. AlCl₃-treated zebrafish larvae. CF, Capsosiphon fulvescens; Normal, untreated CF extracts; CF90, CF subcritical water extract at 90℃ and 3 MPa pressure; CFW, CF water extract; AlCl₃, aluminum chloride.
그림 Fig. 7. Effect of CF90 and CFW on mRNA levels related to neurodegenerative function in AlCl₃-induced zebrafish larvae. The concentration of AlCl₃ was 25 μM. Data are presented as mean ± SE (n = 3) and analyzed by one-way analysis of variance followed by Dunnett test. *p < 0.05, **p < 0.01, ***p < 0.001 vs. untreated normal zebrafish larvae; #p < 0.05, ^##p < 0.01, ^###p < 0.001 vs. AlCl₃-treated zebrafish larvae. CF, Capsosiphon fulvescens; Normal, untreated CF extracts; CF90, CF subcritical water extract at 90℃ and 3 MPa pressure; CFW, CF water extract; AlCl₃, aluminum chloride.
그림 Fig. 8. Effect of CF90 and CFW on mRNA level related to cognitive function in AlCl₃-induced zebrafish larvae. The concentration of AlCl₃ was 25 μM. Data are presented as mean ± SE (n = 3) and analyzed by one-way analysis of variance followed by Dunnett test. *p < 0.05 vs. untreated normal zebrafish larvae; ^#p < 0.05 vs. AlCl₃-treated zebrafish larvae. CF, Capsosiphon fulvescens; Normal, untreated CF extracts; CF90, CF subcritical water extract at 90℃ and 3 MPa pressure; CFW, CF water extract; AlCl₃, aluminum chloride.

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