Neurotoxic Effect of Flavonol Myricetin in the Presence of Excess Copper
Abstract
:1. Introduction
2. Results
2.1. Treatment with Myricetin Exacerbated Copper-Induced Decrease of Viability in SH-SY5Y Cells
2.2. Effects of Myricetin on ROS Generation and GSH Content in the Presence of Excess Copper
2.3. Myricetin Promoted Copper-Induced Changes in Chromatin Condensation and Stimulated LDH Release
2.4. Effects of Copper and Myricetin on the Expression of Proteins Involved in Oxidative Response and Cell Death
2.5. Effects of the Inhibitors of OS-Related Signaling Pathways on Copper and Myricetin-Induced Neuronal Death
2.6. Effects of Myricetin and Copper on the Morphological Properties of SH-SY5Y Cells at the Nanoscale
2.7. Effects of Myricetin and Copper on the Nanomechanical Properties of Neuroblastoma SH-SY5Y Cells
3. Discussion
4. Materials and Methods
4.1. Chemicals
4.2. Culturing of SH-SY5Y Cells
4.3. Drug Treatment
4.4. Assessment of Cell Viability
4.4.1. MTT Assay
4.4.2. Trypan Exclusion Assay
4.4.3. Determination of ATP Level
4.5. Measurement of Lactate Dehydrogenase (LDH) Release from SH-sY5Y Cells Exposed to Copper and Myricetin
4.6. Measurement of Intracellular ROS Accumulation
4.7. Determination of Reduced Glutathione (GSH) Levels
4.8. Determination of Caspase -3/7 Activity
4.9. Nuclear Staining with Hoechst 33342 and Propidium Iodide (PI)
4.10. Western Blot Analysis of PARP-1, p53, p73, Bcl-2, Bax and NME1/2 Protein Expression
4.11. AFM Measurements
4.12. Statistical Analysis
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Treatment | ||||
---|---|---|---|---|
Control Cells | Myricetin (10 μg/mL) | Cu2+ (0.5 mM CuSO4) | Myricetin (10 μg/mL) + Cu2+ (0.5 mM CuSO4) | |
Z range (Ncell = 8) | 353 ± 227 | 389 ± 190 | 514 ± 123 | 540 ± 196 |
Rq/nm (Ncell = 8) | 47 ± 27 | 44 ± 34 | 66 ± 27 | 89 ± 45 |
Ra/nm (Ncell = 8) | 33 ± 16 | 35 ± 28 | 54 ± 23 | 71 ± 35 |
E (Ncel L = 8)/kPa | 4.2 ± 1.1 | 4.5 ± 1.1 | 3.7 ± 1.1 | 2.7 ± 1.3 |
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Sadžak, A.; Vlašić, I.; Kiralj, Z.; Batarelo, M.; Oršolić, N.; Jazvinšćak Jembrek, M.; Kušen, I.; Šegota, S. Neurotoxic Effect of Flavonol Myricetin in the Presence of Excess Copper. Molecules 2021, 26, 845. https://doi.org/10.3390/molecules26040845
Sadžak A, Vlašić I, Kiralj Z, Batarelo M, Oršolić N, Jazvinšćak Jembrek M, Kušen I, Šegota S. Neurotoxic Effect of Flavonol Myricetin in the Presence of Excess Copper. Molecules. 2021; 26(4):845. https://doi.org/10.3390/molecules26040845
Chicago/Turabian StyleSadžak, Anja, Ignacija Vlašić, Zoran Kiralj, Marijana Batarelo, Nada Oršolić, Maja Jazvinšćak Jembrek, Ines Kušen, and Suzana Šegota. 2021. "Neurotoxic Effect of Flavonol Myricetin in the Presence of Excess Copper" Molecules 26, no. 4: 845. https://doi.org/10.3390/molecules26040845