Gardenamide A Protects RGC-5 Cells from H2O2-Induced Oxidative Stress Insults by Activating PI3K/Akt/eNOS Signaling Pathway
Abstract
:1. Introduction
2. Results and Discussion
2.1. GA Dose-Dependently Protected RGC-5 Cells from H2O2-Induced Insults
2.2. GA Protected RGC-5 Cells against Apoptosis Induced by H2O2
2.3. The Protective Effect of GA on RGC-5 Cells Was Mediated by Endothelial Nitrioxide Synthase (eNOS)
Groups | tNOS (U/mL) | cNOS (U/mL) | eNOS (U/mL) | iNOS (U/mL) |
---|---|---|---|---|
ctrl | 8.23 ± 0.43 | 6.42 ± 0.64 | 3.32 ± 0.35 | 1.81 ± 0.33 |
GA | 9.52 ± 0.34 | 8.36 ± 0.52 && | 4.26 ± 0.22 && | 1.16 ± 0.22 |
H2O2 | 5.45 ± 0.22 ** | 1.82 ± 0.19 ** | 1.03 ± 0.17 ** | 3.63 ± 0.42 ** |
H2O2 + GA | 7.51 ± 0.58 ## | 6.20 ± 0.28 ## | 3.14 ± 0.23 ## | 1.31 ± 0.21 ## |
2.4. GA Time- and Dose-Dependently Stimulated the Phosphorylation of ERK1/2 and Akt in RGC-5 Cells
2.5. Neuroprotective Action of GA against H2O2-Induced Impairments to RGC-5 Cells Was Mediated by the Activation of the PI3K/Akt Signaling Pathway
2.6. GA Blocked ROS Production and Lipid Peroxidation Induced by H2O2 in RGC-5 Cells
2.7. Discussion
2.7.1. GA Decreased ROS and MDA Levels in RGC-5 Cells Induced by H2O2
2.7.2. GA Promoted Survival of RGC-5 by Activating eNOS
2.7.3. GA Promoted Survival of RGC-5 by Activation of the PI3K/Akt/eNOS Signaling Pathway
3. Experimental Section
3.1. Chemicals and Reagents
3.2. Cell Culture
3.3. MTT Assay
3.4. Detection of Apoptosis by Hoechst Staining
3.5. Western Blotting
3.6. Mitochondrial Membrane Potential Determination
3.7. Measurement of Intracellular ROS Generation
3.8. Estimation of Lipid Peroxidation
3.9. Determination of NOS Activity
3.10. Data Analysis and Statistics
4. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Wang, R.; Peng, L.; Zhao, J.; Zhang, L.; Guo, C.; Zheng, W.; Chen, H. Gardenamide A Protects RGC-5 Cells from H2O2-Induced Oxidative Stress Insults by Activating PI3K/Akt/eNOS Signaling Pathway. Int. J. Mol. Sci. 2015, 16, 22350-22367. https://doi.org/10.3390/ijms160922350
Wang R, Peng L, Zhao J, Zhang L, Guo C, Zheng W, Chen H. Gardenamide A Protects RGC-5 Cells from H2O2-Induced Oxidative Stress Insults by Activating PI3K/Akt/eNOS Signaling Pathway. International Journal of Molecular Sciences. 2015; 16(9):22350-22367. https://doi.org/10.3390/ijms160922350
Chicago/Turabian StyleWang, Rikang, Lizhi Peng, Jiaqiang Zhao, Laitao Zhang, Cuiping Guo, Wenhua Zheng, and Heru Chen. 2015. "Gardenamide A Protects RGC-5 Cells from H2O2-Induced Oxidative Stress Insults by Activating PI3K/Akt/eNOS Signaling Pathway" International Journal of Molecular Sciences 16, no. 9: 22350-22367. https://doi.org/10.3390/ijms160922350