Abstract
Apoptosis/cell death and reactive oxygen species (ROS) via overload free Ca2+ and Zn2+ uptake into mitochondria are emerging as crucial events in the etiology of hypoxia (HPX)-induced neurodegenerative diseases. The neuroprotective actions of curcumin (CURC) via modulation of oxidative stress and the PARP1-dependent activated TRPM2 cation channel on the ROS generation and cell death in several neurons have been recognized. However, the molecular mechanisms underlying CURC’s neuroprotection remain elusive. We investigated the role of CURC via modulation of TRPM2 on cell death and oxidative cytotoxicity in SH-SY5Y neuronal cells. The SH-SY5Y cells were divided into five groups as follows: CURC (10 µM for 24 h), HPX (200 µM CoCl2 for 24 h), CURC + HPX, and HPX + TRPM2 blockers (2-APB-100 µM or ACA-25 µM for 30 min). In some experiments, the cells in the HPX groups were additionally incubated with PARP1 (PJ34) and Zn2+ (TPEN) inhibitors. The exposure of CoCl2 induced increases of TRPM2 current density and Ca2+ fluorescence intensity with an increase of mitochondrial membrane depolarization and ROS generation. When HPX-induced TRPM2 activity was blocked by 2-APB and ACA, or the cells were treated with CURC, the increase of ROS generation, the expression levels of TRPM2 and PARP1 were restored. The levels of apoptosis and cell death in the cells were enriched with increases of caspase-3 and -9 activations, although they were decreased by CURC treatment. HPX-induced increase of cytosolic Zn2+ was attenuated by the TPEN and CURC treatments. In conclusion, CURC attenuates HPX-induced mitochondrial ROS generation, apoptosis, cell death, and TRPM2-mediated Ca2+ signaling and may provide an avenue for treating HPX-induced neurological diseases associated with the ROS, Ca2+, and Zn2+.
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Abbreviations
- 2-APB:
-
2-Aminoethyl diphenylborinate
- ACA:
-
N-(p-amylcinnamoyl) anthranilic acid
- Ca2 + :
-
Calcium ion
- CURC:
-
Curcumin
- HPX:
-
Hypoxia
- LSM:
-
Laser scan confocal microscope
- MitDep:
-
Mitochondrial membrane depolarization
- ROS:
-
Reactive oxygen species
- TRP:
-
Transient receptor potential
- TRPM2:
-
Transient receptor potential melastatin 2
- Zn2 + :
-
Zinc ion
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Acknowledgments
The authors wish thanks to Prof. Dr. Peter Butterworth (King’s College, London, UK) for polishing English of the manuscript.
Funding
This study was carried out with financial support from BSN Health, Analyses, Innovation, Consultancy, Organization, Agriculture, Industry Ltd. Inc (Göller Bölgesi Teknokenti, Isparta, Turkey). (Project No: 2019-02).
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Professor M Nazıroğlu formulated the present hypothesis and he wrote the main manuscript text. The cell culture, antioxidant, and cell viability analyses in the current study were performed in 4th International Brain Research School, 24–30 June 2019, Isparta, Turkey by Dr. HH Armağan. (http://2019.brs.org.tr/). The authors wish to thanks technicians F Şahin (BSN Health, Analyses, Innovation, Consultancy, Organization, Agriculture, Industry and Trade Limited Company, Göller Bölgesi Teknokenti, Isparta, Turkey) for helping in patch-clamp and Western blot analyses.
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Armağan, H.H., Nazıroğlu, M. Curcumin Attenuates Hypoxia-Induced Oxidative Neurotoxicity, Apoptosis, Calcium, and Zinc Ion Influxes in a Neuronal Cell Line: Involvement of TRPM2 Channel. Neurotox Res 39, 618–633 (2021). https://doi.org/10.1007/s12640-020-00314-w
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DOI: https://doi.org/10.1007/s12640-020-00314-w