Peroxiredoxin 1 alleviates oxygen-glucose deprivation/ reoxygenation injury in N2a cells via suppressing the JNK/caspase-3 pathway

Document Type : Original Article

Authors

1 Department of Anesthesiology, Qingdao Municipal Hospital, Qingdao, Shandong, China

2 Malvern College Qingdao, Qingdao, Shandong, China

3 Department of Anesthesiology, Qingdao Eight People’s Hospital, Qingdao, Shandong, China

4 Department of Anesthesiology, Qingdao Clinical College Affiliated to Nanjing Medical University, Qingdao, Shandong, China

5 Department of Anesthesiology, Weifang No.2 People’s Hospital, Weifang, Shandong, China

Abstract

Objective(s): Cerebral ischemia/reperfusion (I/R) injury inevitably aggravates the initial cerebral tissue damage following a stroke. Peroxiredoxin 1 (Prdx1) is a representative protein of the endogenous antioxidant enzyme family that regulates several reactive oxygen species (ROS)-dependent signaling pathways, whereas the JNK/caspase-3 proapoptotic pathway has a prominent role during cerebral I/R injury. This study aimed to examine the potential mechanism of Prdx1 in Neuro 2A (N2a) cells following oxygen–glucose deprivation and reoxygenation (OGD/R) injury. 
Materials and Methods: N2a cells were exposed to OGD/R to simulate cerebral I/R injury. Prdx1 siRNA transfection and the JNK inhibitor (SP600125) were used to interfere with their relative expressions. CCK-8 assay, flow cytometry, and lactate dehydrogenase (LDH) assay were employed to determine the viability and apoptosis of N2a cells. The intracellular ROS content was assessed using ROS Assay Kit. Real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western blot analyses were conducted to detect the expression levels of Prdx1, JNK, phosphorylated JNK (p-JNK), and cleaved caspase-3. 
Results: Firstly, Prdx1, p-JNK, and cleaved caspase-3 expression were significantly induced in OGD/R-exposed N2a cells. Secondly, the knockdown of Prdx1 inhibited cell viability and increased apoptosis rate, expression of p-JNK, and cleaved caspase-3 expression. Thirdly, SP600125 inhibited the JNK/caspase-3 signaling pathway and mitigated cell injury following OGD/R. Finally, SP600125 partially reversed Prdx1 down-regulation-mediated cleaved caspase-3 activation and OGD/R damage in N2a cells. 
Conclusion: Prdx1 alleviates the injury to N2a cells induced by OGD/R via suppressing JNK/caspase-3 pathway, showing promise as a potential therapeutic for cerebral I/R injury.

Keywords

Main Subjects

References

1. Feske SK. Ischemic Stroke. Am J Med 2021; 134:1457-1464.
2. Kim J, Thayabaranathan T, Donnan GA, Howard G, Howard VJ, Rothwell PM, et al. Global stroke statistics 2019. Int J Stroke 2020; 15:819-838.
3. Xiong XY, Liu L, Yang QW. Refocusing neuroprotection in cerebral reperfusion era: New challenges and strategies. Front Neurol 2018; 9:1-11.
4. Sun MS, Jin H, Sun X, Huang S, Zhang FL, Guo ZN, et al. Free radical damage in ischemia-reperfusion injury: An obstacle in acute ischemic stroke after revascularization therapy. Oxid Med Cell Longev 2018; 2018:3804979.
5. Rodriguez C, Agulla J, Delgado-Esteban M. Refocusing the brain: New approaches in neuroprotection against ischemic injury. Neurochem Res 2021; 46:51-63.
6. Granger DN, Kvietys PR. Reperfusion injury and reactive oxygen species: The evolution of a concept. Redox Biol 2015; 6:524-551.
7. Ledgerwood EC, Marshall JW, Weijman JF. The role of peroxiredoxin 1 in redox sensing and transducing. Arch Biochem Biophys 2017; 617:60-67.
8. Wu J, Chen Y, Yu S, Li L, Zhao X, Li Q, et al. Neuroprotective effects of sulfiredoxin-1 during cerebral ischemia/reperfusion oxidative stress injury in rats. Brain Res Bull 2017; 132:99-108.
9. de Los Reyes Corrales T, Losada-Perez M, Casas-Tinto S. JNK pathway in CNS pathologies. Int J Mol Sci 2021; 22:1-12.
10. Ma S, Liu X, Cheng B, Jia Z, Hua H, Xin Y. Chemical characterization of polysaccharides isolated from scrophularia ningpoensis and its protective effect on the cerebral ischemia/reperfusin injury in rat model. Int J Biol Macromol 2019; 139:955-966.
11. Xia P, Zhang F, Yuan Y, Chen C, Huang Y, Li L, et al. ALDH 2 conferred neuroprotection on cerebral ischemic injury by alleviating mitochondria-related apoptosis through JNK/caspase-3 signing pathway. Int J Biol Sci 2020; 16:1303-1323.
12. Guo W, Liu X, Li J, Shen Y, Zhou Z, Wang M, et al. Prdx1 alleviates cardiomyocyte apoptosis through ROS-activated MAPK pathway during myocardial ischemia/reperfusion injury. Int J Biol Macromol 2018; 112:608-615.
13. Wu R, Yun Q, Zhang J, Wang Z, Zhang X, Bao J. Knockdown of circular RNA tousled-like kinase 1 relieves ischemic stroke in middle cerebral artery occlusion mice and oxygen-glucose deprivation and reoxygenation-induced N2a cell damage. Bioengineered 2022; 13:3434-3449.
14. Wang H, Yang H, Chang M, Sun F, Qi H, Li X. Long non-coding RNA TTTY15 sponges miR-520a-3p to exacerbate neural apoptosis induced by cerebral ischemia/reperfusion via targeting IRF9 both in vivo and in vitro. Biomed J 2022; 46:100530.
15. Zhu D, Fang C, Yang Z, Ren Y, Yang F, Zheng S, et al. Tubulin-binding peptide RR-171 derived from human umbilical cord serum displays antitumor activity against hepatocellular carcinoma via inducing apoptosis and activating the NF-kappa B pathway. Cell Prolif 2022; 55:e13241.
16. Lv WP, Li MX, Wang L. Peroxiredoxin 1 inhibits lipopolysaccharide-induced oxidative stress in lung tissue by regulating P38/JNK signaling pathway. Eur Rev Med Pharmacol Sci 2017; 21:1876-1883.
17. Duan LH, Li M, Wang CB, Wang QM, Liu QQ, Shang WF, et al. Protective effects of organic extracts of alpinia oxyphylla against hydrogen peroxide-induced cytotoxicity in PC12 cells. Neural Regen Res 2020; 15:682-689.
18. Su Y, Feng W, Shi J, Chen L, Huang J, Lin T. circRIP2 accelerates bladder cancer progression via miR-1305/Tgf-beta2/smad3 pathway. Mol Cancer 2020; 19:23.
19. Mathew B, Ravindran S, Liu X, Torres L, Chennakesavalu M, Huang CC, et al. Mesenchymal stem cell-derived extracellular vesicles and retinal ischemia-reperfusion. Biomaterials 2019; 197:146-160.
20. Warach SJ, Dula AN, Milling TJ, Jr. Tenecteplase Thrombolysis for acute ischemic stroke. Stroke 2020; 51:3440-3451.
21. Wu D, Li M, Fisher M, Ji X. Brain cytoprotection of ischemic stroke in the era of effective reperfusion. Sci Bull (Beijing) 2022; 67:2372-2375.
22. Shen L, Gan Q, Yang Y, Reis C, Zhang Z, Xu S, et al. Mitophagy in cerebral ischemia and ischemia/reperfusion injury. Front Aging Neurosci 2021; 13:687246.
23. Mao R, Zong N, Hu Y, Chen Y, Xu Y. Neuronal Death mechanisms and therapeutic strategy in ischemic stroke. Neurosci Bull 2022; 38:1229-1247.
24. Jurcau A, Simion A. Neuroinflammation in cerebral ischemia and ischemia/reperfusion injuries: from pathophysiology to therapeutic strategies. Int J Mol Sci 2021; 23:1-31.
25. Sun K, Fan J, Han J. Ameliorating effects of traditional Chinese medicine preparation, Chinese materia medica and active compounds on ischemia/reperfusion-induced cerebral microcirculatory disturbances and neuron damage. Acta Pharm Sin B 2015; 5:8-24.
26. Solassol J, Crozet C, Lehmann S. Prion propagation in cultured cells. Br Med Bull 2003; 66:87-97.
27. Orellana-Urzua S, Rojas I, Libano L, Rodrigo R. Pathophysiology of ischemic stroke: Role of oxidative stress. Curr Pharm Des 2020; 26:4246-4260.
28. Fu X, Wang J, Liao S, Lv Y, Xu D, Yang M, et al. (1)H NMR-Based metabolomics reveals refined-huang-lian-jie-du-decoction (BBG) as a potential ischemic stroke treatment drug with efficacy and a favorable therapeutic window. Front Pharmacol 2019; 10:1-17.
29. Chen W, Li D. Reactive oxygen species (ROS)-responsive nanomedicine for solving ischemia-reperfusion injury. Front Chem 2020; 8:1-7.
30. Yuan Q, Yuan Y, Zheng Y, Sheng R, Liu L, Xie F, et al. Anti-cerebral ischemia reperfusion injury of polysaccharides: A review of the mechanisms. Biomed Pharmacother 2021; 137:111303.
31. Chen CH, Hsieh CL. Effect of acupuncture on oxidative stress induced by cerebral ischemia-reperfusion injury. Anti-oxidants (Basel) 2020; 9:1-15.
32. Min Y, Kim MJ, Lee S, Chun E, Lee KY. Inhibition of TRAF6 ubiquitin-ligase activity by PRDX1 leads to inhibition of NFKB activation and autophagy activation. Autophagy 2018; 14:1347-1358.
33. Cui S, Wang C, Bai W, Li J, Pan Y, Huang X, et al. CD1d1 intrinsic signaling in macrophages controls NLRP3 inflammasome expression during inflammation. Sci Adv 2020; 6:eaaz7290.
34. Jeong SJ, Kim S, Park JG, Jung IH, Lee MN, Jeon S, et al. Prdx1 (peroxiredoxin 1) deficiency reduces cholesterol efflux via impaired macrophage lipophagic flux. Autophagy 2018; 14:120-133.
35. Jeong SJ, Park JG, Oh GT. Peroxiredoxins as potential targets for cardiovascular disease. Anti-oxidants (Basel) 2021; 10:1-15.
36. Burnside SW, Hardingham GE. Transcriptional regulators of redox balance and other homeostatic processes with the potential to alter neurodegenerative disease trajectory. Biochem Soc Trans 2017; 45:1295-1303.
37. Lee YM, Park SH, Shin DI, Hwang JY, Park B, Park YJ, et al. Oxidative modification of peroxiredoxin is associated with drug-induced apoptotic signaling in experimental models of Parkinson disease. J Biol Chem 2008; 283:9986-9998.
38. Yang GQ, Huang JC, Yuan JJ, Zhang Q, Gong CX, Chen Q, et al. Prdx1 reduces intracerebral hemorrhage-induced brain injury via targeting inflammation- and apoptosis-related mRNA stability. Front Neurosci 2020; 14:181.
39. Li L, Zhu K, Liu Y, Wu X, Wu J, Zhao Y, et al. Targeting thioredoxin-1 with siRNA exacerbates oxidative stress injury after cerebral ischemia/reperfusion in rats. Neuroscience 2015; 284:815-823.
40. Wang M, Zhu K, Zhang L, Li L, Zhao J. Thioredoxin 1 protects astrocytes from oxidative stress by maintaining peroxiredoxin activity. Mol Med Rep 2016; 13:2864-2870.
41. Gomez-Osuna A, Calatrava V, Galvan A, Fernandez E, Llamas A. Identification of the MAPK cascade and its relationship with nitrogen metabolism in the green alga chlamydomonas reinhardtii. Int J Mol Sci 2020; 21:1-21.
42. Nijboer CH, van der Kooij MA, van Bel F, Ohl F, Heijnen CJ, Kavelaars A. Inhibition of the JNK/AP-1 pathway reduces neuronal death and improves behavioral outcome after neonatal hypoxic-ischemic brain injury. Brain Behav Immun 2010; 24:812-821.
43. Hu W, Wu X, Yu D, Zhao L, Zhu X, Li X, et al. Regulation of JNK signaling pathway and RIPK3/AIF in necroptosis-mediated global cerebral ischemia/reperfusion injury in rats. Exp Neurol 2020; 331:113374.
44. Shvedova M, Anfinogenova Y, Atochina-Vasserman EN, Schepetkin IA, Atochin DN. c-Jun N-Terminal Kinases (JNKs) in myocardial and cerebral ischemia/reperfusion injury. Front Pharmacol 2018; 9:1-18.
45. Zheng J, Dai Q, Han K, Hong W, Jia D, Mo Y, et al. JNK-IN-8, a c-Jun N-terminal kinase inhibitor, improves functional recovery through suppressing neuroinflammation in ischemic stroke. J Cell Physiol 2020; 235:2792-2799.
46. Kim YJ, Lee WS, Ip C, Chae HZ, Park EM, Park YM. Prx1 suppresses radiation-induced c-Jun NH2-terminal kinase signaling in lung cancer cells through interaction with the glutathione S-transferase Pi/c-Jun NH2-terminal kinase complex. Cancer Res 2006; 66:7136-7142.
47. Ishii T, Warabi E, Yanagawa T. Novel roles of peroxiredoxins in inflammation, cancer and innate immunity. J Clin Biochem Nutr 2012; 50:91-105.
48. Kim SU, Park YH, Kim JM, Sun HN, Song IS, Huang SM, et al. Dominant role of peroxiredoxin/JNK axis in stemness regulation during neurogenesis from embryonic stem cells. Stem Cells 2014; 32:998-1011.
49. Hopkins BL, Nadler M, Skoko JJ, Bertomeu T, Pelosi A, Shafaei PM, et al. A peroxidase peroxiredoxin 1-specific redox regulation of the novel FOXO3 microRNA target let-7. Antioxid Redox Signal 2018; 28:62-77.
50. Soto D, Gomez-Serrano M, Pieralisi A, Calvo JC, Peral B, Guerra LN. N-acetylcysteine inhibits kinase phosphorylation during 3T3-L1 adipocyte differentiation. Redox Rep 2017; 22:265-271.
Iranian Journal of Basic Medical Sciences
Volume 26, Issue 11
November 2023
Pages 1305-1312

Files

Share

How to cite

Statistics