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Norepinephrine Reduces Reactive Oxygen Species (ROS) and DNA Damage in Ovarian Surface Epithelial Cells.

Patel PR, Hegde ML, Theruvathu J, Mitra SA, Boldogh I, Sowers L - J Bioanal Biomed (2015)

Bottom Line: Treatment with NE resulted in significantly less ROS generation than in untreated cells (p<0.01).NE treatment after hydrogen peroxide treatment resulted in a noticeable decrease in ROS generation.This effect is protective in the presence of the oxidative-damaging agent bleomycin.

View Article: PubMed Central - PubMed

Affiliation: Department of Obstetrics and Gynecology, The University of Texas Medical Branch in Galveston, Texas, USA.

ABSTRACT

Objective: To determine the role of norepinephrine (NE) on DNA damage and reactive oxygen species (ROS) generation in ovarian surface epithelial cells.

Method: Non-tumorigenic, immortalized ovarian surface epithelial cells were treated with NE, bleomycin, and bleomycin followed by NE. The comet assay was performed on each treatment group to determine the amount of single and double-strand breaks induced by treatments. ROS levels for each treatment group were measured using the H2DCF-DA fluorescence assay. Finally, RNA transcripts were measured for each treatment group with regards to the expression of DNA repair and oxidative stress genes.

Results: The mean tail moment of untreated cells was significantly greater than that of cells treated with NE (p=0.02). The mean tail moment of cells treated with bleomycin was significantly greater than that of cells treated with bleomycin followed by NE (p<0.01). Treatment with NE resulted in significantly less ROS generation than in untreated cells (p<0.01). NE treatment after hydrogen peroxide treatment resulted in a noticeable decrease in ROS generation. Genes associated with oxidative stress were upregulated in cells treated with bleomycin, however this upregulation was blunted when bleomycin-treated cells were treated subsequently with NE.

Conclusion: NE is associated with decreased DNA damage and ROS production in ovarian surface epithelial cells. This effect is protective in the presence of the oxidative-damaging agent bleomycin. These results suggest an additional physiologic role for the stress hormone NE, in protecting ovarian surface epithelial cells from oxidative stress.

No MeSH data available.


Related in: MedlinePlus

ROS generation measured by H2DCFDA. As expected, treatment with hydrogen peroxide resulted in greater ROS generation. Treatment with norepinephrine resulted in significantly less ROS generation than in untreated cells (p<0.01). Norepinephrine treatment after hydrogen peroxide treatment resulted in decreased ROS generation, however this different was not significant (p=0.14). H2O2: Hydrogen Peroxide; NE: Norepinephrine.
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Figure 4: ROS generation measured by H2DCFDA. As expected, treatment with hydrogen peroxide resulted in greater ROS generation. Treatment with norepinephrine resulted in significantly less ROS generation than in untreated cells (p<0.01). Norepinephrine treatment after hydrogen peroxide treatment resulted in decreased ROS generation, however this different was not significant (p=0.14). H2O2: Hydrogen Peroxide; NE: Norepinephrine.

Mentions: ROS data is summarized in Figure 4. Treatment with NE resulted in decreased ROS, as exhibited by decreased fluorescence emission throughout the 50 min experimental period. In addition, when comparing the treatment with H2O2 with the treatment with H2O2 followed by NE, both groups initially had similar fluorescence patterns until the NE was added to the second group, resulting in an abrupt decrease in fluorescence. There was significantly less ROS generation when cells were treated with NE compared to untreated cells (p<0.01). There tended to be less ROS generation after treatment with H2O2 followed by NE compared to treatment with only H2O2 (p=0.14).


Norepinephrine Reduces Reactive Oxygen Species (ROS) and DNA Damage in Ovarian Surface Epithelial Cells.

Patel PR, Hegde ML, Theruvathu J, Mitra SA, Boldogh I, Sowers L - J Bioanal Biomed (2015)

ROS generation measured by H2DCFDA. As expected, treatment with hydrogen peroxide resulted in greater ROS generation. Treatment with norepinephrine resulted in significantly less ROS generation than in untreated cells (p<0.01). Norepinephrine treatment after hydrogen peroxide treatment resulted in decreased ROS generation, however this different was not significant (p=0.14). H2O2: Hydrogen Peroxide; NE: Norepinephrine.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4495967&req=5

Figure 4: ROS generation measured by H2DCFDA. As expected, treatment with hydrogen peroxide resulted in greater ROS generation. Treatment with norepinephrine resulted in significantly less ROS generation than in untreated cells (p<0.01). Norepinephrine treatment after hydrogen peroxide treatment resulted in decreased ROS generation, however this different was not significant (p=0.14). H2O2: Hydrogen Peroxide; NE: Norepinephrine.
Mentions: ROS data is summarized in Figure 4. Treatment with NE resulted in decreased ROS, as exhibited by decreased fluorescence emission throughout the 50 min experimental period. In addition, when comparing the treatment with H2O2 with the treatment with H2O2 followed by NE, both groups initially had similar fluorescence patterns until the NE was added to the second group, resulting in an abrupt decrease in fluorescence. There was significantly less ROS generation when cells were treated with NE compared to untreated cells (p<0.01). There tended to be less ROS generation after treatment with H2O2 followed by NE compared to treatment with only H2O2 (p=0.14).

Bottom Line: Treatment with NE resulted in significantly less ROS generation than in untreated cells (p<0.01).NE treatment after hydrogen peroxide treatment resulted in a noticeable decrease in ROS generation.This effect is protective in the presence of the oxidative-damaging agent bleomycin.

View Article: PubMed Central - PubMed

Affiliation: Department of Obstetrics and Gynecology, The University of Texas Medical Branch in Galveston, Texas, USA.

ABSTRACT

Objective: To determine the role of norepinephrine (NE) on DNA damage and reactive oxygen species (ROS) generation in ovarian surface epithelial cells.

Method: Non-tumorigenic, immortalized ovarian surface epithelial cells were treated with NE, bleomycin, and bleomycin followed by NE. The comet assay was performed on each treatment group to determine the amount of single and double-strand breaks induced by treatments. ROS levels for each treatment group were measured using the H2DCF-DA fluorescence assay. Finally, RNA transcripts were measured for each treatment group with regards to the expression of DNA repair and oxidative stress genes.

Results: The mean tail moment of untreated cells was significantly greater than that of cells treated with NE (p=0.02). The mean tail moment of cells treated with bleomycin was significantly greater than that of cells treated with bleomycin followed by NE (p<0.01). Treatment with NE resulted in significantly less ROS generation than in untreated cells (p<0.01). NE treatment after hydrogen peroxide treatment resulted in a noticeable decrease in ROS generation. Genes associated with oxidative stress were upregulated in cells treated with bleomycin, however this upregulation was blunted when bleomycin-treated cells were treated subsequently with NE.

Conclusion: NE is associated with decreased DNA damage and ROS production in ovarian surface epithelial cells. This effect is protective in the presence of the oxidative-damaging agent bleomycin. These results suggest an additional physiologic role for the stress hormone NE, in protecting ovarian surface epithelial cells from oxidative stress.

No MeSH data available.


Related in: MedlinePlus