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Glutamine Reduces the Apoptosis of H9C2 Cells Treated with High-Glucose and Reperfusion through an Oxidation-Related Mechanism.

Li K, Cui YC, Zhang H, Liu XP, Zhang D, Wu AL, Li JJ, Tang Y - PLoS ONE (2015)

Bottom Line: Data indicated that high glucose and hypoxia-reoxygenation were associated with a dramatic decline of intercellular glutamine and increase in apoptosis.Glutamine supplementation was also associated with less S-glutathionylation and increased the activity of complex I, leading to less mitochondrial ROS formation.We conclude that apoptosis induced by high glucose and hypoxia-reoxygenation was reduced by glutamine supplementation, via decreased oxidative stress and inactivation of the intrinsic apoptotic pathway.

View Article: PubMed Central - PubMed

Affiliation: Animal Experimental Center, Beijing Key Laboratory of Preclinical Research and Evaluation for Cardiovascular Implant Materials, State key Laboratory of Cardiovascular Disease, Fu Wai Hospital, National Center for Cardiovascular Disease, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.

ABSTRACT
Mitochondrial overproduction of reactive oxygen species (ROS) in diabetic hearts during ischemia/reperfusion injury and the anti-oxidative role of glutamine have been demonstrated. However, in diabetes mellitus the role of glutamine in cardiomyocytes during ischemia/reperfusion injury has not been explored. To examine the effects of glutamine and potential mechanisms, in the present study, rat cardiomyoblast H9C2 cells were exposed to high glucose (33 mM) and hypoxia-reoxygenation. Cell viability, apoptosis, intracellular glutamine, and mitochondrial and intracellular glutathione were determined. Moreover, ROS formation, complex I activity, membrane potential and adenosine triphosphate (ATP) content were also investigated. The levels of S-glutathionylated complex I and mitochondrial apoptosis-related proteins, including cytochrome c and caspase-3, were analyzed by western blot. Data indicated that high glucose and hypoxia-reoxygenation were associated with a dramatic decline of intercellular glutamine and increase in apoptosis. Glutamine supplementation correlated with a reduction in apoptosis and increase of glutathione and glutathione reduced/oxidized ratio in both cytoplasm and mitochondria, but a reduction of intracellular ROS. Glutamine supplementation was also associated with less S-glutathionylation and increased the activity of complex I, leading to less mitochondrial ROS formation. Furthermore, glutamine supplementation prevented from mitochondrial dysfunction presented as mitochondrial membrane potential and ATP levels and attenuated cytochrome c release into the cytosol and caspase-3 activation. We conclude that apoptosis induced by high glucose and hypoxia-reoxygenation was reduced by glutamine supplementation, via decreased oxidative stress and inactivation of the intrinsic apoptotic pathway.

No MeSH data available.


Related in: MedlinePlus

Effect of Gln on mitochondrial ROS formation.H9C2 cells were treated as described in Fig 3. (A) Representative imaging of Mito Sox confocal image of each group. (B) Mean Mito Sox fluorescence intensities are presented as percentage relative to the control value. The data are expressed as the means ±S.D. of 3 independent experiments. * P < 0.05 compared with the control group; # P < 0.05 compare with the 1 mM and 4 mM Gln groups. a.u. = arbitrary units.
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pone.0132402.g005: Effect of Gln on mitochondrial ROS formation.H9C2 cells were treated as described in Fig 3. (A) Representative imaging of Mito Sox confocal image of each group. (B) Mean Mito Sox fluorescence intensities are presented as percentage relative to the control value. The data are expressed as the means ±S.D. of 3 independent experiments. * P < 0.05 compared with the control group; # P < 0.05 compare with the 1 mM and 4 mM Gln groups. a.u. = arbitrary units.

Mentions: We next examined complex I activity and mtROS production in the control and HG+H/R-treated groups. Treatment of the HG and H/R significantly decreased the activity of complex I (P < 0.05, Fig 4F). The complex I activity in the 1 mM, 4 mM, and 16 mM Gln groups was 44%, 46%, and 61% that of the control group, respectively. Mitochondria-derived ROS levels in the HG + H/R groups were significantly higher than that of the control group (P < 0.05, Fig 5A and 5B). However, the increase was not observed in the 16 mM Gln group (P < 0.05).


Glutamine Reduces the Apoptosis of H9C2 Cells Treated with High-Glucose and Reperfusion through an Oxidation-Related Mechanism.

Li K, Cui YC, Zhang H, Liu XP, Zhang D, Wu AL, Li JJ, Tang Y - PLoS ONE (2015)

Effect of Gln on mitochondrial ROS formation.H9C2 cells were treated as described in Fig 3. (A) Representative imaging of Mito Sox confocal image of each group. (B) Mean Mito Sox fluorescence intensities are presented as percentage relative to the control value. The data are expressed as the means ±S.D. of 3 independent experiments. * P < 0.05 compared with the control group; # P < 0.05 compare with the 1 mM and 4 mM Gln groups. a.u. = arbitrary units.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4493145&req=5

pone.0132402.g005: Effect of Gln on mitochondrial ROS formation.H9C2 cells were treated as described in Fig 3. (A) Representative imaging of Mito Sox confocal image of each group. (B) Mean Mito Sox fluorescence intensities are presented as percentage relative to the control value. The data are expressed as the means ±S.D. of 3 independent experiments. * P < 0.05 compared with the control group; # P < 0.05 compare with the 1 mM and 4 mM Gln groups. a.u. = arbitrary units.
Mentions: We next examined complex I activity and mtROS production in the control and HG+H/R-treated groups. Treatment of the HG and H/R significantly decreased the activity of complex I (P < 0.05, Fig 4F). The complex I activity in the 1 mM, 4 mM, and 16 mM Gln groups was 44%, 46%, and 61% that of the control group, respectively. Mitochondria-derived ROS levels in the HG + H/R groups were significantly higher than that of the control group (P < 0.05, Fig 5A and 5B). However, the increase was not observed in the 16 mM Gln group (P < 0.05).

Bottom Line: Data indicated that high glucose and hypoxia-reoxygenation were associated with a dramatic decline of intercellular glutamine and increase in apoptosis.Glutamine supplementation was also associated with less S-glutathionylation and increased the activity of complex I, leading to less mitochondrial ROS formation.We conclude that apoptosis induced by high glucose and hypoxia-reoxygenation was reduced by glutamine supplementation, via decreased oxidative stress and inactivation of the intrinsic apoptotic pathway.

View Article: PubMed Central - PubMed

Affiliation: Animal Experimental Center, Beijing Key Laboratory of Preclinical Research and Evaluation for Cardiovascular Implant Materials, State key Laboratory of Cardiovascular Disease, Fu Wai Hospital, National Center for Cardiovascular Disease, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.

ABSTRACT
Mitochondrial overproduction of reactive oxygen species (ROS) in diabetic hearts during ischemia/reperfusion injury and the anti-oxidative role of glutamine have been demonstrated. However, in diabetes mellitus the role of glutamine in cardiomyocytes during ischemia/reperfusion injury has not been explored. To examine the effects of glutamine and potential mechanisms, in the present study, rat cardiomyoblast H9C2 cells were exposed to high glucose (33 mM) and hypoxia-reoxygenation. Cell viability, apoptosis, intracellular glutamine, and mitochondrial and intracellular glutathione were determined. Moreover, ROS formation, complex I activity, membrane potential and adenosine triphosphate (ATP) content were also investigated. The levels of S-glutathionylated complex I and mitochondrial apoptosis-related proteins, including cytochrome c and caspase-3, were analyzed by western blot. Data indicated that high glucose and hypoxia-reoxygenation were associated with a dramatic decline of intercellular glutamine and increase in apoptosis. Glutamine supplementation correlated with a reduction in apoptosis and increase of glutathione and glutathione reduced/oxidized ratio in both cytoplasm and mitochondria, but a reduction of intracellular ROS. Glutamine supplementation was also associated with less S-glutathionylation and increased the activity of complex I, leading to less mitochondrial ROS formation. Furthermore, glutamine supplementation prevented from mitochondrial dysfunction presented as mitochondrial membrane potential and ATP levels and attenuated cytochrome c release into the cytosol and caspase-3 activation. We conclude that apoptosis induced by high glucose and hypoxia-reoxygenation was reduced by glutamine supplementation, via decreased oxidative stress and inactivation of the intrinsic apoptotic pathway.

No MeSH data available.


Related in: MedlinePlus