Kurkumin Yüksek Doz Morfinin Neden Olduğu Apoptozis ve Oksidatif Nörotoksisiteyi TRPV4 Katyon Kanalını Düzenleyerek Azaltır

Öz

Amaç: Nöropatik ağrıyı tedavi etmek için yüksek doz (H-MRP) ve uzun süreli morfin uygulanması kullanılır. Bununla birlikte, H-MRP tedavisi, morfin toleransı, aşırı reaktif oksijen türleri (ROS) üretimi, apoptozis ve Ca+2 akışının artmasına neden olur. Sinir hasarı nedeniyle zarar gören SH-SY5Y sinir hücrelerinde, kurkumin (CRC) TRPV4 katyon kanalını inhibe ederek ROS'un neden olduğu apoptozis artışını azaltır. H-MRP, SH-SY5Y hücrelerinde TRPV4 katyon kanalını inhibe ederek aşırı ROS ve apoptozis oluşumunu önlemek için CRC etkisi henüz araştırılmamıştır. Bu çalışmada, CRC tedavisinin TRPV4 kanalını düzenleyerek mitokondriyal ROS üretimini ve apoptozis oluşumunu H-MRP inkübasyonunu nasıl etkilediğini SH-SY5Y hücrelerinde araştırıldı. Materyal ve Metot: SH-SY5Y hücrelerinde dört ana grup oluşturuldu. Bunlar; kontrol, normal morfin (N-MRP) (50 µM ve 24 saat), H-MRP (500 µM ve 24 saat) ve H-MRP + CRC (5 µM ve 24 saat). Bulgular: TRPV4 agonisti (GSK1016790A) ile inkübasyon, ROS, mitokondriyal fonksiyon bozukluğu, debris (ölü hücre artığı), apoptosis, kaspaz -3, kaspaz -8, kaspaz -9 ve lipit peroksidasyon düzeylerini artırdı. Bununla birlikte, TRPV4 antagonisti (rutenyum kırmızısı) ve CRC inkübasyonları bu artışları azalttı. Rutenyum kırmızısı ve CRC inkübasyonları, H-MRP inkübasyonunun neden olduğu hücre canlılığı yüzdesi, canlı hücre sayısı, glutatyon ve glutatyon peroksidaz düzeyleri azalışlarını artırdı. Sonuç: CRC tedavisi, TRPV4 kanalını baskılayarak H-MRP'nin mitokondriyal oksidan ve sinir hücre ölümü etkilerini azalttı. H-MRP neden olduğu mitokondriyal oksidatif stres ve sinir hücre harabiyetini önlemek için CRC tedavisi alternatif bir kaynak tedavi olarak gözükmektedir.

Anahtar Kelimeler

• Kurkumin
• sinir hasarı
• mitokondriyal oksidatif stres
• morfin
• TRPV4 kanalı

Curcumin Reduces High-dose Morphine-Induced Apoptosis and Oxidative Neurotoxicity via TRPV4 Cation Channel Suppression

Abstract

Objective: Long-term and high-dose morphine (H-MRP) treatments for neuropathic pain cause the body to become extremely susceptible to morphine tolerance, which increases the amount of toxic reactive oxygen species (ROS), apoptosis, and calcium (Ca2+) entering the neuron. It has been known that curcumin (CRC) decreased these increases in ROS-damaged SH-SY5Y cells by blocking the TRPV4 cation channel. It has not been studied whether CRC can also suppress the high levels of ROS and apoptosis caused by H-MRP in SH-SY5Y cells by affecting TRPV4. So, the study was carried out to investigate whether CRC can suppress the high level of mitochondrial ROS and apoptosis. Materials and Methods: In the SH-SY5Y, four primary groups were induced as control, normal morphine (N-MRP) (50 µM for 24h), H-MRP (500 µM for 24h), H-MRP + CRC (5 µM for 24h). Results: While the incubations of TRPV4 antagonist (ruthenium red) and CRC decreased the H-MRP-induced increases of apoptosis, caspase-3, caspase-8, caspase-9, ROS, mitochondrial dysfunction, debris number, and lipid peroxidation levels, the TRPV4 agonist (GSK1016790A) stimulation further increased these levels. The CRC increased glutathione, glutathione peroxidase, live cell number, and cell viability percentage, all of which were decreased by H-MRP. Conclusions: The levels of H-MRP-induced neuronal death and mitochondrial oxidative stress were reduced by CRC treatment through TRPV4 inhibition. For H-MRP-induced mitochondrial oxidative neuronal injury, CRC is a potential treatment option.

Keywords

• Curcumin
• neuronal injury
• mitochondrial oxidative stress
• morphine
• TRPV4 channel

References

1. Badshah I, Anwar M, Murtaza B, Khan MI. Molecular mechanisms of morphine tolerance and dependence; novel insights and future perspectives. Mol Cell Biochem. 2024;479(6):1457-1485. doi:10.1007/s11010-023-04810-3
2. Su LY, Liu Q, Jiao L, Yao YG. Molecular mechanism of neuroprotective effect of melatonin on morphine addiction and analgesic tolerance: an update. Mol Neurobiol. 2021;58(9):4628-4638. doi:10.1007/s12035-021-02448-0  
3. Kuthati Y, Wong CS. The melatonin type 2 receptor agonist IIK7 attenuates and reverses morphine tolerance in neuropathic pain rats through the suppression of neuroinflammation in the spinal cord. Pharmaceuticals (Basel). 2024;17(12):1638. doi:10.3390/ph17121638
4. Amiri R, Fallah F, Ghorbanzadeh B, Oroojan AA, Behmanesh MA, Alboghobeish S. Mitigating morphine dependence and withdrawal: The role of venlafaxine and calcium channel blockers in mitochondrial damage and oxidative stress in the brain. Brain Res Bull. 2025;226:111364. doi:10.1016/j.brainresbull.2025.111364  
5. Amini K, Zhaleh H, Tahvilian R. Effects of low-dose morphine suppress methamphetamine-induced cell death by inhibiting the ROS generation and caspase-3 activity. Bratisl Lek Listy. 2019;120(5):336-343. doi:10.4149/BLL_2019_055
6. Sheikholeslami MA, Parvardeh S, Ghafghazi S, Sabetkasaei M. Curcumin attenuates morphine dependence by modulating μ-opioid receptors and glial cell-activated neuroinflammation in rat. Neuropeptides. 2023;98:102318. doi:10.1016/j.npep.2022.102318
7. Kharazmi K, Alani B, Heydari A, Ardjmand A. Protection against morphine-induced inhibitory avoidance memory impairment in rat by curcumin: Possible role of nitric oxide/ cAMP-response element binding protein pathway. Iran J Med Sci. 2022;47(6):594-602. doi:10.30476/IJMS.2022
8. Motaghinejad M, Karimian M, Motaghinejad O, Shabab B, Yazdani I, Fatima S. Protective effects of various dosage of Curcumin against morphine induced apoptosis and oxidative stress in rat isolated hippocampus. Pharmacol Rep. 2015;67(2):230-5. doi: 10.1016/j.pharep.2014.09.006

9. Hu X, Huang F, Szymusiak M, Liu Y, Wang ZJ. Curcumin attenuates opioid tolerance and dependence by inhibiting Ca2+/calmodulin-dependent protein kinase II α activity. J Pharmacol Exp Ther. 2015;352(3):420-8. doi: 10.1124/jpet.114.219303  
10. Panda P, Mohanty S, Gouda SR, et al. Advanced strategies for enhancing the neuroprotective potential of curcumin: delivery systems and mechanistic insights in neurodegenerative disorders. Nutr Neurosci. 2025:1-26. doi:10.1080/1028415X.2025.2472773
11. Osmanlıoğlu HÖ, Yıldırım MK, Akyuva Y, Yıldızhan K, Nazıroğlu M. Morphine induces apoptosis, inflammation, and mitochondrial oxidative stress via activation of TRPM2 channel and nitric oxide signaling pathways in the hippocampus. Mol Neurobiol. 2020;57(8):3376-3389. doi:10.1007/s12035-020-01975-6
12. Nazıroğlu M. A novel antagonist of TRPM2 and TRPV4 channels: Carvacrol. Metab Brain Dis. 2022;37(3):711-728. doi: 10.1007/s11011-021-00887-1
13. Çınar R, Yıldızhan K. Curcumin protects against MPP+-induced neurotoxicity in SH-SY5Y cells by modulating the TRPV4 channel. Mol Biol Rep. 2025;52(1):255. doi:10.1007/s11033-025-10345-1  
14. Özşimşek A, Nazıroğlu M. The involvement of TRPV4 on the hypoxia-induced oxidative neurotoxicity and apoptosis in a neuronal cell line: Protective role of melatonin. Neurotoxicology. 2021;87:136-148. doi:10.1016/j.neuro.2021.09.003  
15. Sánchez JC, Muñoz LV, Ehrlich BE. Modulating TRPV4 channels with paclitaxel and lithium. Cell Calcium. 2020;91:102266. doi:10.1016/j.ceca.2020.102266
16. Osmanlıoğlu HÖ, Nazıroğlu M. Resveratrol modulates diabetes-induced neuropathic pain, apoptosis, and oxidative neurotoxicity in mice through TRPV4 channel inhibition. Mol Neurobiol. 2024;61(9):7269-7286. doi:10.1007/s12035-024-04311-4
17. Armağan HH, 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. 2021;39(3):618-633. doi:10.1007/s12640-020-00314-w
18. Rullo L, Caputi FF, Losapio LM, et al. Effects of different opioid drugs on oxidative status and proteasome activity in SH-SY5Y cells. Molecules. 2022;27(23):8321. doi:10.3390/molecules27238321
19. Gu F, Zhou Y, Tian L, et al. Morphine promotes non-small cell lung cancer progression by downregulating E-cadherin via the PI3K/AKT/mTOR pathway. Sci Rep. 2024;14(1):21130. doi:10.1038/s41598-024-72198-1
20. Ning J, Chen X, Li Q, et al. Bidirectional effects of morphine on pancreatic cancer progression via the p38/JNK pathway. Sci Rep. 2024;14(1):24233. doi:10.1038/s41598-024-75089-7
21. Osmanlıoğlu HÖ. Propofol's Neuroprotective Effect against cisplatin-induced oxidative neurotoxicity via suppression of the TRPM2 cation channel. Online Turkish Journal of Health Sciences 2024;9(3):254-262 2024;9(3):254-262. doi:10.26453/otjhs.1532340
22. Ciltas AC, Ozdemir E, Gunes H, Ozturk A. Inhibition of the TRPM2 cation channel attenuates morphine tolerance by modulating endoplasmic reticulum stress and apoptosis in rats. Neurosci Lett. 2025;851:138168. doi:10.1016/j.neulet.2025.138168
23. Lipscombe D, Lopez-Soto EJ. Epigenetic control of ion channel expression and cell-specific splicing in nociceptors: Chronic pain mechanisms and potential therapeutic targets. Channels (Austin). 2021;15(1):156-164. doi:10.1080/19336950.2020.1860383  
24. Ke S, Dong P, Mei Y, et al. A synthetic peptide, derived from neurotoxin GsMTx4, acts as a non-opioid analgesic to alleviate mechanical and neuropathic pain through the TRPV4 channel. Acta Pharm Sin B. 2025;15(3):1447-1462. doi:10.1016/j.apsb.2024.12.028  
25. Shao J, Han J, Zhu Y, et al. Curcumin induces endothelium-dependent relaxation by activating endothelial TRPV4 channels. J Cardiovasc Transl Res. 2019;12(6):600-607. doi:10.1007/s12265-019-09928-8
26. Nazıroğlu M. A mini review of curcumin and TRPM2 channel: Focus on oxidative neurotoxicity. J Cell Neurosci Oxid Stress 2022;14(3): 1105-1112. doi:10.37212/jcnos.1325009
27. Reddy AC, Lokesh BR. Effect of dietary turmeric (Curcuma longa) on iron-induced lipid peroxidation in the rat liver. Food Chem Toxicol. 1994;32(3):279-83. doi:10.1016/0278-6915(94)90201-1  
28. Al-Rubaei ZM, Mohammad TU, Ali LK. Effects of local curcumin on oxidative stress and total antioxidant capacity in vivo study. Pak J Biol Sci. 2014;17(12):1237-41. doi:10.3923/pjbs.2014.1237.1241