Effect of rutin on oxidative DNA damage in PC12 neurons cultured in nutrients deprivation condition

Document Type: Original Article

Authors

1 Department of Pharmacology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran

2 Pharmacological Research Center of Medicinal Plants, Mashhad University of Medical Sciences, Mashhad, Iran

3 Department of Pharmacology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

4 Anesthesiology and Critical Care Research Center, Shiraz University of Medical Sciences, Shiraz, Iran

5 Division of Neurocognitive Sciences, Psychiatry and Behavioral Sciences Research Center, Mashhad, University of Medical Sciences, Mashhad, Iran

Abstract

Objective(s): Rutin is a flavonoid with potent antioxidant property, which exhibited cytoprotective effects in several models of neuronal injury. This work aimed to examine whether rutin can protect neurons against oxidative DNA damage caused by serum/glucose deprivation (SGD) as an in vitro model of neurodegeneration and ischemia.
Materials and Methods: The PC12 cells were cultured for 2 hr in normal culture medium containing different concentrations of rutin or α-tocopherol (positive control) and then further incubated for 12 hr in SGD condition. Then, cell viability, DNA fragmentation, lipid peroxidation, generation of reactive oxygen species (ROS), and the expression of proteins involved in apoptosis were determined.
Results: The SGD condition significantly decreased viability of the cells, which was accompanied by a significant rise in the generation of ROS and lipid peroxidation. Rutin enhanced the viability of PC12 cells in SGD condition and reduced the production of ROS and lipid peroxidation. In addition, rutin decreased DNA damage and inhibited apoptotic cell death by decreasing the levels of proapoptotic proteins (Bax, caspase-3, caspase-9) and increasing the level of anti-apoptotic protein Bcl-2.
Conclusion: This study demonstrated that rutin inhibits oxidative DNA damage and neuronal death induced by nutrients deprivation condition. Further studies may warrant the use of rutin as an appropriate neuroprotective agent for ischemic attacks and other neurodegenerative disorders.

Keywords


1. Doyle KP, Simon RP, Stenzel-Poore MP. Mechanisms of ischemic brain damage. Neuropharmacology 2008; 55:310-318.
2. Li P, Stetler RA, Leak RK, Shi Y, Li Y, Yu W, et al. Oxidative stress and DNA damage after cerebral ischemia: Potential therapeutic targets to preserve the genome and improve stroke recovery. Neuropharmacology 2018:134(Pt B):208-217.
3. Chauhan A, Chauhan V. Oxidative stress in autism. Pathophysiology 2006; 13:171-181.
4. Chamorro Á, Dirnagl U, Urra X, Planas AM. Neuroprotection in acute stroke: targeting excitotoxicity, oxidative and nitrosative stress, and inflammation. Lancet Neurol 2016; 15:869-881.
5. Na J-Y, Song K, Kim S, Kwon J. Rutin protects rat articular chondrocytes against oxidative stress induced by hydrogen peroxide through SIRT1 activation. Biochem Biophys Res Commun 2016; 473:1301-1308.
6. Singh S, Singh DK, Meena A, Dubey V, Masood N, Luqman S. Rutin protects t‑butyl hydroperoxide-induced oxidative impairment via modulating the Nrf2 and iNOS activity. Phytomedicine. 2019; 55:92-104.
7. Gao M, Ma Y, Liu D. Rutin suppresses palmitic acids-triggered inflammation in macrophages and blocks high fat diet-induced obesity and fatty liver in mice. Pharm Res 2013; 30:2940-2950.
8. Sadeghnia HR, Yousefsani BS, Rashidfar M, Boroushaki MT, Asadpour E, Ghorbani A. Protective effect of rutin on hexachlorobutadiene-induced nephrotoxicity. Renal Failure 2013; 35:1151-1155.
9. Ghorbani A. Mechanisms of antidiabetic effects of flavonoid rutin. Biomed Pharmacother 2017; 96:305-312.
10. Nassiri-Asl M, Shariati-Rad S, Zamansoltani F. Anticonvulsive effects of intracerebroventricular administration of rutin in rats. Prog Neuro-Psychopharmacol Biol Psychiatry 2008; 32:989-993.
11. Xu P-x, Wang S-w, Yu X-l, Su Y-j, Wang T, Zhou W-w, et al. Rutin improves spatial memory in Alzheimer’s disease transgenic mice by reducing Aβ oligomer level and attenuating oxidative stress and neuroinflammation. Behav Brain Res 2014; 264:173-180.
12. Song H-l, Zhang X, Wang W-z, Liu R-h, Zhao K, Liu M-y, et al. Neuroprotective mechanisms of rutin for spinal cord injury through anti-oxidation and anti-inflammation and inhibition of p38 mitogen activated protein kinase pathway. Neural Regen Res 2018; 13:128-134.
13. Hao G, Dong Y, Huo R, Wen K, Zhang Y, Liang G. Rutin inhibits neuroinflammation and provides neuroprotection in an experimental rat model of subarachnoid hemorrhage, possibly through suppressing the RAGE–NF-κB inflammatory signaling pathway. Neurochem Res 2016; 41:1496-1504.
14. Khan MM, Ahmad A, Ishrat T, Khuwaja G, Srivastawa P, Khan MB, et al. Rutin protects the neural damage induced by transient focal ischemia in rats. Brain Res 2009; 1292:123-135.
15. Pu F, Mishima K, Irie K, Motohashi K, Tanaka Y, Orito K, et al. Neuroprotective effects of quercetin and rutin on spatial memory impairment in an 8-arm radial maze task and neuronal death induced by repeated cerebral ischemia in rats. J Pharmacol Sci 2007; 104:329-334.
16. Hillion JA, Takahashi K, Maric D, Ruetzler C, Barker JL, Hallenbeck JM. Development of an ischemic tolerance model in a PC12 cell line. J Cereb Blood Flow Metab 2005; 25:154-162.
17. Mousavi SH, Bakhtiari E, Hosseini A, Jamialahmadi K. Protective effects of glucosamine and its acetylated derivative on serum/glucose deprivation-induced PC12 cells death: Role of reactive oxygen species. Res Pharm Sci 2018; 13:121-129.
18. Jin Y, Tang X, Cao X, Yu L, Chen J, Zhao H, et al. 4-((5-(Tert-butyl)-3-chloro-2-hydroxybenzyl) amino)-2-hydroxybenzoic acid protects against oxygen-glucose deprivation/reperfusion injury. Life Sci 2018: 204:46-54.
19. Lin W-C, Peng Y-F, Hou C-W. Ferulic acid protects PC12 neurons against hypoxia by inhibiting the p-MAPKs and COX-2 pathways. Iran J Basic Med Sci 2015; 18:478-484.
20. Asadpour E, Ghorbani A, Sadeghnia HR. Water-soluble compounds of lettuce inhibit DNA damage and lipid peroxidation induced by glucose/serum deprivation in N2a cells. Acta Pol Pharm 2014; 71:409-413.
21. Hadjzadeh M, Tavakol Afshari J, Ghorbani A, Shakeri M. The effects of aqueous extract of garlic (Allium sativum L.) on laryngeal cancer cells (Hep-2) and L929 cells in vitro. J Med Plants 2006; 2:41-48.
22. Sadeghnia HR, Kamkar M, Assadpour E, Boroushaki MT, Ghorbani A. Protective effect of safranal, a constituent of Crocus sativus, on quinolinic acid-induced oxidative damage in rat hippocampus. Iran J Basic Med Sci 2013; 16:73-82.
23. Prince PSM, Priya S. Preventive effects of rutin on lysosomal enzymes in isoproterenol induced cardio toxic rats: Biochemical, histological and in vitro evidences. Eur J Pharmacol 2010; 649:229-235.
24. Koda T, Kuroda Y, Imai H. Protective effect of rutin against spatial memory impairment induced by trimethyltin in rats. Nutr Res 2008; 28:629-634.
25. Broughton BR, Reutens DC, Sobey CG. Apoptotic mechanisms after cerebral ischemia. Stroke 2009; 40:e331-e339.
26. Bhat AH, Dar KB, Anees S, Zargar MA, Masood A, Sofi MA, et al. Oxidative stress, mitochondrial dysfunction and neurodegenerative diseases; a mechanistic insight. Biomed Pharmacother 2015; 74:101-110.
27. Park S-E, Sapkota K, Choi J-H, Kim M-K, Kim YH, Kim KM, et al. Rutin from Dendropanax morbifera Leveille protects human dopaminergic cells against rotenone induced cell injury through inhibiting JNK and p38 MAPK signaling. Neurochem Res 2014; 39:707-718.
28. Ündeğer Ü, Aydın S, Başaran AA, Başaran N. The modulating effects of quercetin and rutin on the mitomycin C induced DNA damage. Toxicol Lett 2004; 151:143-149.
29. Muhammad A, Arthur DE, Babangida S, Erukainure OL, Malami I, Sani H, et al. Modulatory role of rutin on 2,5-hexanedione-induced chromosomal and DNA damage in rats: validation of computational predictions. Drug Chem Toxicol 2018: 1-14.