Nat Prod Sci.  2018 Sep;24(3):148-154. 10.20307/nps.2018.24.3.148.

Acer okamotoanum Inhibit the Hydrogen Peroxide-Induced Oxidative Stress in C6 Glial Cells

• Choi SY
• Kim JH
• Quilantang NG
• Lee S
• Cho EJ

• KMID: 2422005
• DOI: http://doi.org/10.20307/nps.2018.24.3.148

Abstract

Chronic oxidative stress due to the accumulation of reactive oxygen species (ROS) in neuronal cells ultimately leads to neurodegenerative diseases. The use of natural therapies for the prevention of ROS-induced cell damage and for the treatment of neurodegenerative disorders has shown promising results. In this study, we evaluated the neuroprotective effects of the ethyl acetate (EtOAc) fraction of A. Okamotoanum against the hydrogen peroxide (H₂O₂)-induced oxidative stress in C6 glial cells. Results show that cell viability was decreased in cells incubated with H₂O₂, whereas the addition of EtOAc fraction treatments in such cells significantly increased viability. The EtOAc fraction showed the highest inhibitory activity against ROS production and it also decreased the expressions of inflammatory proteins including cyclooxygenase-2, inducible nitric oxide synthase and interleukin-1β. Furthermore, the EtOAc fraction inhibited apoptosis by regulating the protein expressions cleaved caspase -9, -3, poly ADP ribose polymerase, Bax and Bcl-2. Therefore, these results show that the EtOAc fraction of A. Okamotoanum exhibits neuroprotective effects against H₂O₂ induced oxidative damage by regulating the inflammatory reaction and apoptotic pathway.

Keyword

Acer okamotoanum; C6 glial cell; Inflammation; Apoptosis; Reactive oxygen species

MeSH Terms

Acer*
Apoptosis
Cell Survival
Cyclooxygenase 2
Hydrogen Peroxide
Hydrogen*
Inflammation
Neurodegenerative Diseases
Neuroglia*
Neurons
Neuroprotective Agents
Nitric Oxide Synthase Type II
Oxidative Stress*
Poly(ADP-ribose) Polymerases
Reactive Oxygen Species
Cyclooxygenase 2
Hydrogen
Hydrogen Peroxide
Neuroprotective Agents
Nitric Oxide Synthase Type II
Poly(ADP-ribose) Polymerases
Reactive Oxygen Species

Figure

• 

  Fig. 1 The effects of the EtOAc fraction of A. okamotoanum on the cell viability of H2O2-treated C6 glial cells. Cells were pretreated with different concentrations (5, 25, 50, 100 µg/mL) of the EtOAc fraction of A. okamotoanum for 2 hr, and then incubated with 300 µM H2O2 for 24 hr. Values are shown as mean ± SD. a~e Means; different letters are significantly different (P<0.05) using the Duncan's multiple range test.

• 

  Fig. 2 The effects of the EtOAc fraction of A. okamotoanum on the ROS levels of H2O2-treated C6 glial cells. Cells were pretreated with different concentrations (5, 25, 50, 100 µg/mL) of the EtOAc fraction of A. okamotoanum for 2 hr, and then incubated with 300 µM H2O2 for 24 hr. Values are shown as mean ± SD. a~c Means; different letters are significantly different (P<0.05) using the Duncan's multiple range test.

• 

  Fig. 3 The effects of the EtOAc fraction of A. okamotoanum on the expression of IκB-α, Cox-2, iNOS and IL-1β in H2O2-treated C6 glial cells. Cells were pretreated with different concentrations (2.5, 5, 25 µg/mL) of the EtOAc fraction of A. okamotoanum for 2 hr, and then incubated with 300 µM H2O2 for 24 hr. β-actin was used as loading control. Values are shown as the mean ± SD. a~e Means; different letters are significantly different (P<0.05) using the Duncan's multiple range test.

• 

  Fig. 4 The effects of the EtOAc fraction of A. okamotoanum on Bax and Bcl-2 expression (A) and the ratio of Bax/Bcl-2 (B) in H2O2-treated C6 glial cells. Cells were pretreated with different concentrations (2.5, 5, 25 µg/mL) of the EtOAc fraction of A. okamotoanum for 2 hr, and then incubated with 300 µM H2O2 for 24 hr. β-actin was used as loading control. Values are shown as the mean ± SD. a~e Means; different letters are significantly different (P<0.05) using the Duncan's multiple range test.

• 

  Fig. 5 The effects of the EtOAc fraction of A. okamotoanumon H2O2-induced increases cleaved caspase 9 (A), cleaved caspase 3 (B), and cleaved PARP (C) levels in C6 glial cells. Cells were pretreated with different concentrations (2.5, 5, 25 µg/mL) of the EtOAc fraction of A. okamotoanum for 2 hr, and then incubated with 300 µM H2O2 for 24 hr. β-actin was used as loading control. Values are shown as the mean ± SD. a~eMeans; different letters are significantly different (P<0.05) using the Duncan's multiple range test.

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