Role of exogenous putrescine in the status of energy, DNA damage, inflammation, and spermidine/spermine-n(1)- acetyltransferase in brain ischemia-reperfusion in rats

Document Type : Original Article


1 Department of Pharmacology, Faculty of Medicine, Selcuk University, Konya, Turkey

2 Department of Physiology, Faculty of Medicine, Selcuk University, Konya, Turkey

3 Department of Biochemistry, Faculty of Medicine, Selcuk University, Konya, Turkey


Objective(s): This study aims to investigate the role of putrescine against brain ischemia-reperfusion (IR) injured rats administered with 250 µmol/kg exogenous putrescine and highlight the IR-associated mechanisms in energy metabolism and inflammatory pathway. 
Materials and Methods: The rats were divided into six groups: 1-Sham group; 2-IR group, 30 min of ischemia and 30 min of reperfusion was performed with bilateral carotid occlusion (BCAO); 3-IPR group, a single oral dose of putrescine was administered at the start of the 30-minute reperfusion; while in the other treatment groups, 4 doses of putrescine were given within 12-hour intervals. After 30 min of reperfusion, the first dose was administered immediately in the IR-PI (group 4), after 3 hr in IR-PII (group 5), and after 6 hr in IR-PIII (group 6). Interleukin-6 (IL-6), Nuclear factor NF-kappa-B (NF-kB), Adenosine triphosphate (ATP), total Nitric oxide (NO), 8-hydroxyguanosine (8-OHdG), Spermidine/Spermin N-acetyltransferase (SSAT) levels were analyzed in brain tissues.
Results: IR reduced brain ATP levels; however, putrescine treatment reversed this state. Brain NO and 8-OHdG levels, and NF-kB and IL-6 levels increased significantly in the IR group and these elevations were decreased in putrescine administered groups. SSAT levels were higher in the IR-PII group. The lowest levels were observed in the IR-PIII group. 
Conclusion: The exogenous putrescine supplementation after cerebral IR creates neuroprotective effects independent of the time of administration; according to conditions such as formation of radicals in the brain, the spread of the inflammation and the need for consumption of energy are considered as a whole. 


1. Cetin N, Suleyman H, Sener E, Demirci E, Gundogdu C, Akcay F. The prevention of ischemia/reperfusion induced oxidative damage by venous blood in rabbit kidneys monitored with biochemical, histopatological and immunohistochemical analysis. J Physiol Pharmacol 2014; 65:383-392.
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 repair the genome and improve stroke recovery. Neuropharmacology 2018; 134:208-217.
3. Girotti AW. Lipid hydroperoxide generation, turnover, and effector action in biological systems. J Lipid Res 1998; 39:1529-1542.
4. Semenza GL. Cellular and molecular dissection of reperfusion injury: ROS within and without. Circ Res 2000; 86:117-118.
5. Subedi L, Gaire BP. Phytochemicals as regulators of microglia/macrophages activation in cerebral ischemia. Pharmacol Res 2021:105419.
6. Tabor CW, Tabor H. Polyamines. Annu Rev Biochem 1984; 53:749-790.
7. Casero RA, Stewart TM, Pegg AE. Polyamine metabolism and cancer: treatments, challenges and opportunities. Nat Rev Cancer 2018; 18:681-695.
8. Liu J-H, Wang T-W, Lin Y-Y, Ho W-C, Tsai H-C, Chen S-P, et al. Acrolein is involved in ischemic stroke-induced neurotoxicity through spermidine/spermine-N1-acetyltransferase activation. Exp Neurol 2020; 323:113066.
9. Pegg AE. Spermidine/spermine-N(1)-acetyltransferase: a key metabolic regulator. Am J Physiol Endocrinol Metab 2008; 294:E995-1010.
10. Zoli M, Pedrazzi P, Zini I, Agnati LF. Spermidine/spermine N1-acetyltransferase mRNA levels show marked and region-specific changes in the early phase after transient forebrain ischemia. Brain Res Mol Brain Res 1996; 38:122-134.
11. Rao AM, Hatcher JF, Doğan A, Dempsey RJ. Elevated N1-acetylspermidine levels in gerbil and rat brains after CNS injury. J Neurochem 2000; 74:1106-1111.
12. Behroozi-Lak T, Zarei L, Moloody-Tapeh M, Farhad N, Mohammadi R. Protective effects of intraperitoneal administration of nimodipine on ischemia-reperfusion injury in ovaries: Histological and biochemical assessments in a rat model. J Pediatr Surg 2017; 52:602-608.
13. ArunaDevi R, Lata S, Bhadoria BK, Ramteke VD, Kumar S, Sankar P, et al. Neuroprotective effect of 5,7,3’,4’,5’-pentahydroxy dihydroflavanol-3-O-(2’’-O-galloyl)-beta-D-glucopyranoside, a polyphenolic compound in focal cerebral ischemia in rat. Eur J Pharmacol 2010; 626:205-212.
14. Cai F, Li C, Wu J, Min Q, Ouyang C, Zheng M, et al. Modulation of the oxidative stress and nuclear factor kappaB activation by theaflavin 3,3’-gallate in the rats exposed to cerebral ischemia-reperfusion. Folia Biol (Praha) 2007; 53:164-172.
15. Dhara M, Matta JA, Lei M, Knowland D, Yu H, Gu S, et al. Polyamine regulation of ion channel assembly and implications for nicotinic acetylcholine receptor pharmacology. Nat Commun 2020; 11:2799.
16. Neuman MG. Biomarkers of Drug-Induced Liver Toxicity. Ther Drug Monit 2019; 41:227-234.
17. Oz M, Demir EA, Caliskan M, Mogulkoc R, Baltaci AK, Nurullahoglu Atalik KE. 3’,4’-Dihydroxyflavonol attenuates spatial learning and memory impairments in global cerebral ischemia. Nutr Neurosci 2017; 20:119-126.
18. Babu GN, Sailor KA, Sun D, Dempsey RJ. Spermidine/spermine N1-acetyl transferase activity in rat brain following transient focal cerebral ischemia and reperfusion. Neurosci Lett 2001; 300:17-20.
19. Okumura S, Teratani T, Fujimoto Y, Zhao X, Tsuruyama T, Masano Y, et al. Oral administration of polyamines ameliorates liver ischemia/reperfusion injury and promotes liver regeneration in rats. Liver Transpl 2016; 22:1231-1244.
20. Caliskan M, Mogulkoc R, Baltaci AK, Menevse E. The effect of 3′, 4′-dihydroxyflavonol on lipid peroxidation in rats with cerebral ischemia reperfusion injury. Neuroche Res 2016; 41:1732-1740.
21. Baines CP. The mitochondrial permeability transition pore and ischemia-reperfusion injury. Basic Res Cardiol 2009; 104:181-188.
22. Vujcic S, Diegelman P, Bacchi CJ, Kramer DL, Porter CW. Identification and characterization of a novel flavin-containing spermine oxidase of mammalian cell origin. Biochem J 2002; 367:665-675.
23. Temiz C, Dogan A, Baskaya MK, Dempsey RJ. Effect of difluoromethylornithine on reperfusion injury after temporary middle cerebral artery occlusion. J Clin Neurosci 2005; 12:449-452.
24. Doğan A, Rao AM, Hatcher J, Rao VL, Başkaya MK, Dempsey RJ. Effects of MDL 72527, a specific inhibitor of polyamine oxidase, on brain edema, ischemic injury volume, and tissue polyamine levels in rats after temporary middle cerebral artery occlusion. J Neurochem 1999; 72:765-770.
25. Kawabori M, Yenari MA. Inflammatory responses in brain ischemia. Curr Med Chem 2015; 22:1258-1277.
26. Clark WM, Rinker LG, Lessov NS, Hazel K, Hill JK, Stenzel-Poore M, et al. Lack of interleukin-6 expression is not protective against focal central nervous system ischemia. Stroke 2000; 31:1715-1720.
27. Yamashita T, Sawamoto K, Suzuki S, Suzuki N, Adachi K, Kawase T, et al. Blockade of interleukin‐6 signaling aggravates ischemic cerebral damage in mice: possible involvement of Stat3 activation in the protection of neurons. J Neurochem 2005; 94:459-468.
28. Herrmann O, Tarabin V, Suzuki S, Attigah N, Coserea I, Schneider A, et al. Regulation of body temperature and neuroprotection by endogenous interleukin-6 in cerebral ischemia. J Cereb Blood Flow Metab 2003; 23:406-415.
29. Dasdelen D, Solmaz M, Menevse E, Mogulkoc R, Baltaci AK, Erdogan E. Increased apoptosis, tumor necrosis factor-α, and DNA damage attenuated by 3’, 4’-dihydroxyflavonol in rats with brain İschemia-reperfusion. Indian J Pharmacol 2021; 53:39.