Limits...
Anandamide protects HT22 cells exposed to hydrogen peroxide by inhibiting CB1 receptor-mediated type 2 NADPH oxidase.

Jia J, Ma L, Wu M, Zhang L, Zhang X, Zhai Q, Jiang T, Wang Q, Xiong L - Oxid Med Cell Longev (2014)

Bottom Line: HT22 cells exposed to H2O2 demonstrated morphological changes, decreased LDH release, reduced metabolic activity, increased levels of intracellular ROS and oxidized glutathione (GSSG), reduced levels of superoxide dismutase (SOD), and reduced glutathione (GSH) and increased expression of Nox2.AEA prevented these effects, a property abolished by simultaneous administration of CB1 antagonist AM251 or CB1-siRNA.Nox2 inhibition is involved in AEA-induced cytoprotection against oxidative stress through CB1 activation in HT22 cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Anesthesiology, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China.

ABSTRACT

Background: Endogenous cannabinoid anandamide (AEA) protects neurons from oxidative injury in rodent models; however the mechanism of AEA-induced neuroprotection remains to be determined. Activation of neuronal NADPH oxidase 2 (Nox2) contributes to oxidative damage of the brain, and inhibition of Nox2 can attenuate cerebral oxidative stress. We aimed to determine whether the neuronal Nox2 was involved in protection mediated by AEA.

Methods: The mouse hippocampal neuron cell line HT22 was exposed to hydrogen peroxide (H2O2) to mimic oxidative injury of neurons. The protective effect of AEA was assessed by measuring cell metabolic activity, apoptosis, lactate dehydrogenase (LDH) release, cellular morphology, intracellular reactive oxygen species (ROS), and antioxidant and oxidant levels and Nox2 expression.

Results: HT22 cells exposed to H2O2 demonstrated morphological changes, decreased LDH release, reduced metabolic activity, increased levels of intracellular ROS and oxidized glutathione (GSSG), reduced levels of superoxide dismutase (SOD), and reduced glutathione (GSH) and increased expression of Nox2. AEA prevented these effects, a property abolished by simultaneous administration of CB1 antagonist AM251 or CB1-siRNA.

Conclusion: Nox2 inhibition is involved in AEA-induced cytoprotection against oxidative stress through CB1 activation in HT22 cells.

Show MeSH

Related in: MedlinePlus

AEA increased the metabolic activity of HT22 cells exposed to H2O2 in a dose-dependent manner. (a) The correlation between the H2O2 concentration and cell metabolic activity. HT22 cells were exposed to different concentrations of H2O2 for 3 h (n = 8). (b) AEA increased the cell metabolic activity of HT22 cells exposed to 200 μM H2O2 for 3 h (n = 8). Results are expressed as means ± SD, *P < 0.05, ***P < 0.001 versus the control (no H2O2, and no AEA), #P < 0.05 versus the cells exposed to H2O2 alone.
© Copyright Policy - open-access
Related In: Results  -  Collection


getmorefigures.php?uid=PMC4127243&req=5

fig2: AEA increased the metabolic activity of HT22 cells exposed to H2O2 in a dose-dependent manner. (a) The correlation between the H2O2 concentration and cell metabolic activity. HT22 cells were exposed to different concentrations of H2O2 for 3 h (n = 8). (b) AEA increased the cell metabolic activity of HT22 cells exposed to 200 μM H2O2 for 3 h (n = 8). Results are expressed as means ± SD, *P < 0.05, ***P < 0.001 versus the control (no H2O2, and no AEA), #P < 0.05 versus the cells exposed to H2O2 alone.

Mentions: HT22 cells were exposed to H2O2 for 3 h, which decreased the cell metabolic activity in a dose-dependent manner. Exposure to 200 μM H2O2 decreased the cell metabolic activity by roughly 50% (Figure 2(a)), and we used this condition for the subsequent experiments.


Anandamide protects HT22 cells exposed to hydrogen peroxide by inhibiting CB1 receptor-mediated type 2 NADPH oxidase.

Jia J, Ma L, Wu M, Zhang L, Zhang X, Zhai Q, Jiang T, Wang Q, Xiong L - Oxid Med Cell Longev (2014)

AEA increased the metabolic activity of HT22 cells exposed to H2O2 in a dose-dependent manner. (a) The correlation between the H2O2 concentration and cell metabolic activity. HT22 cells were exposed to different concentrations of H2O2 for 3 h (n = 8). (b) AEA increased the cell metabolic activity of HT22 cells exposed to 200 μM H2O2 for 3 h (n = 8). Results are expressed as means ± SD, *P < 0.05, ***P < 0.001 versus the control (no H2O2, and no AEA), #P < 0.05 versus the cells exposed to H2O2 alone.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4127243&req=5

fig2: AEA increased the metabolic activity of HT22 cells exposed to H2O2 in a dose-dependent manner. (a) The correlation between the H2O2 concentration and cell metabolic activity. HT22 cells were exposed to different concentrations of H2O2 for 3 h (n = 8). (b) AEA increased the cell metabolic activity of HT22 cells exposed to 200 μM H2O2 for 3 h (n = 8). Results are expressed as means ± SD, *P < 0.05, ***P < 0.001 versus the control (no H2O2, and no AEA), #P < 0.05 versus the cells exposed to H2O2 alone.
Mentions: HT22 cells were exposed to H2O2 for 3 h, which decreased the cell metabolic activity in a dose-dependent manner. Exposure to 200 μM H2O2 decreased the cell metabolic activity by roughly 50% (Figure 2(a)), and we used this condition for the subsequent experiments.

Bottom Line: HT22 cells exposed to H2O2 demonstrated morphological changes, decreased LDH release, reduced metabolic activity, increased levels of intracellular ROS and oxidized glutathione (GSSG), reduced levels of superoxide dismutase (SOD), and reduced glutathione (GSH) and increased expression of Nox2.AEA prevented these effects, a property abolished by simultaneous administration of CB1 antagonist AM251 or CB1-siRNA.Nox2 inhibition is involved in AEA-induced cytoprotection against oxidative stress through CB1 activation in HT22 cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Anesthesiology, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China.

ABSTRACT

Background: Endogenous cannabinoid anandamide (AEA) protects neurons from oxidative injury in rodent models; however the mechanism of AEA-induced neuroprotection remains to be determined. Activation of neuronal NADPH oxidase 2 (Nox2) contributes to oxidative damage of the brain, and inhibition of Nox2 can attenuate cerebral oxidative stress. We aimed to determine whether the neuronal Nox2 was involved in protection mediated by AEA.

Methods: The mouse hippocampal neuron cell line HT22 was exposed to hydrogen peroxide (H2O2) to mimic oxidative injury of neurons. The protective effect of AEA was assessed by measuring cell metabolic activity, apoptosis, lactate dehydrogenase (LDH) release, cellular morphology, intracellular reactive oxygen species (ROS), and antioxidant and oxidant levels and Nox2 expression.

Results: HT22 cells exposed to H2O2 demonstrated morphological changes, decreased LDH release, reduced metabolic activity, increased levels of intracellular ROS and oxidized glutathione (GSSG), reduced levels of superoxide dismutase (SOD), and reduced glutathione (GSH) and increased expression of Nox2. AEA prevented these effects, a property abolished by simultaneous administration of CB1 antagonist AM251 or CB1-siRNA.

Conclusion: Nox2 inhibition is involved in AEA-induced cytoprotection against oxidative stress through CB1 activation in HT22 cells.

Show MeSH
Related in: MedlinePlus