NLRP3 Inflammasome Is Involved in Q-VD-OPH Induced Necroptosis Following Cerebral Ischemia-Reperfusion Injury
- 52 Downloads
Necroptosis is a manner of caspase-independent cell death,which accounts for delayed ischemic cerebral injury, and can be used as a novel tool to expand the treatment time window in ischemic cerebral injury. Q-VD-OPH, a novel pan caspase inhibitor, has been identified as an inducer of necroptosis. In this study, we determined the optimal dose of Q-VD-OPH, which induces necroptosis in rats by the middle cerebral artery occlusion, followed by reperfusion. Furthermore, we report that the NLRP3 inflammasome is involved in necroptosis, with levels of NLRP3 inflammasome proteins as well as inflammatory cytokines, such as IL-1β, being elevated. We also demonstrated that NLRP3 was not only expressed in microglia and vascular endothelial cell, but also in neurons when necroptosis is induced with Q-VD-OPH. Inhibition of NLRP3 by glyburide strongly suppressed the expression of NLRP3 inflammasome proteins and IL-1β, and markedly reduced brain tissue damage. Our findings provide evidence that pretreatment with Q-VD-OPH suppresses apoptosis and induces necroptosis in the cerebral ischemia-reperfusion model. We also identified that the NLRP3 inflammasome plays an important role in neuronal necroptosis, and that NLRP3 inflammasome deficiency reduces brain tissue damage after cerebral ischemia-reperfusion injury in rats.
KeywordsNLRP3 inflammasome Q-VD-OPH Necroptosis Apoptosis Brain ischemia
This study was supported by Grants 81501138 from the National Natural Science Foundation of China and KC15SH077 from Xuzhou Science and Technology Project. We would like to thank Hongyan Dong, Ting Li and Fuxing Dong from Laboratory of Neurobiology Research Center of Xuzhou Medical University for advices and help.
Compliance with Ethical Standards
Conflict of interest
There is no conflict of interest to disclose.
- 2.Degterev A, Huang Z, Boyce M, Li Y, Jagtap P, Mizushima N, Cuny GD, Mitchison TJ, Moskowitz MA, Yuan J (2005) Chemical inhibitor of nonapoptotic cell death with therapeutic potential for ischemic brain injury. Nat Chem Biol 1(2):112–119. https://doi.org/10.1038/nchembio711 CrossRefPubMedGoogle Scholar
- 10.Antar V, Akdemir O, Sağmanligil A, Sahan E, Çelik Ö, Çolak A, Karaoğlan A (2009) Q-VD-OPh, a pancaspase inhibitor, reduces trauma-induced apoptosis and improves the recovery of hind-limb function in rats after spinal cord injury. Neurocirugia 20(6):533–540 (discussion 540)CrossRefPubMedGoogle Scholar
- 13.Renolleau S, Fau S, Goyenvalle C, Joly LM, Chauvier D, Jacotot E, Mariani J, Charriaut-Marlangue C (2007) Specific caspase inhibitor Q-VD-OPh prevents neonatal stroke in P7 rat: a role for gender. J Neurochem 100(4):1062–1071. https://doi.org/10.1111/j.1471-4159.2006.04269.x CrossRefPubMedGoogle Scholar
- 15.Fann DY, Lee SY, Manzanero S, Tang SC, Gelderblom M, Chunduri P, Bernreuther C, Glatzel M, Cheng YL, Thundyil J, Widiapradja A (2013) Intravenous immunoglobulin suppresses NLRP1 and NLRP3 inflammasome-mediated neuronal death in ischemic stroke. Cell Death Dis 4:e790. https://doi.org/10.1038/cddis.2013.326 CrossRefPubMedGoogle Scholar
- 19.Hou X, Yang C, Zhang L, Hu T, Sun D, Cao H, Yang F, Guo G, Gong C, Zhang X, Tong A (2017) Killing colon cancer cells through PCD pathways by a novel hyaluronic acid-modified shell-core nanoparticle loaded with RIP3 in combination with chloroquine. Biomaterials 124:195–210. https://doi.org/10.1016/j.biomaterials.2016.12.032 CrossRefPubMedGoogle Scholar
- 21.Mohagheghi F, Ahmadiani A, Rahmani B, Moradi F, Romond N, Khalaj L (2013) Gemfibrozil pretreatment resulted in a sexually dimorphic outcome in the rat models of global cerebral ischemia-reperfusion via modulation of mitochondrial pro-survival and apoptotic cell death factors as well as MAPKs. J Mol Neurosci 50(3):379–393. https://doi.org/10.1007/s12031-012-9932-0 CrossRefPubMedGoogle Scholar
- 33.Wang JY, Xia Q, Chu KT, Pan J, Sun LN, Zeng B, Zhu YJ, Wang Q, Wang K, Luo BY (2011) Severe global cerebral ischemia-induced programmed necrosis of hippocampal CA1 neurons in rat is prevented by 3-methyladenine: a widely used inhibitor of autophagy. J Neuropathol Experim Neurol 70(4):314–322. https://doi.org/10.1097/NEN.0b013e31821352bd CrossRefGoogle Scholar
- 34.Schwamm LH, Ali SF, Reeves MJ, Smith EE, Saver JL, Messe S, Bhatt DL, Grau-Sepulveda MV, Peterson ED, Fonarow GC (2013) Temporal trends in patient characteristics and treatment with intravenous thrombolysis among acute ischemic stroke patients at get with the guidelines-stroke hospitals. Circ-Cardiovasc Qual 6(5):543–549CrossRefGoogle Scholar
- 36.Garcia-Bermejo P, Calleja AI, Perez-Fernandez S, Cortijo E, del Monte JM, Garcia-Porrero M, Fe Muñoz M, Fernández-Herranz R, Arenillas JF (2012) Perfusion computed tomography-guided intravenous thrombolysis for acute ischemic stroke beyond 4.5 hours: a case-control study. Cerebrovasc Dis 34(1):31–37CrossRefPubMedGoogle Scholar
- 38.Saleh D, Najjar M, Zelic M, Shah S, Nogusa S, Polykratis A, Paczosa MK, Gough PJ, Bertin J, Whalen M, Fitzgerald KA (2017) Kinase activities of RIPK1 and RIPK3 can direct IFN-beta synthesis induced by lipopolysaccharide. J Immunol 198(11):4435–4447. https://doi.org/10.4049/jimmunol.1601717 CrossRefPubMedPubMedCentralGoogle Scholar
- 39.Oliva-Martin MJ, Sanchez-Abarca LI, Rodhe J, Carrillo-Jimenez A, Vlachos P, Herrera AJ, Garcia-Quintanilla A, Caballero-Velazquez T, Perez-Simon JA, Joseph B, Venero JL (2016) Caspase-8 inhibition represses initial human monocyte activation in septic shock model. Oncotarget 7(25):37456–37470. https://doi.org/10.18632/oncotarget.9648 CrossRefPubMedPubMedCentralGoogle Scholar
- 43.Savage CD, Lopez-Castejon G, Denes A, Brough D (2012) NLRP3-inflammasome activating DAMPs stimulate an inflammatory response in glia in the absence of priming which contributes to brain inflammation after injury. Front Immunol. https://doi.org/10.3389/Fimmu.2012.00288 PubMedPubMedCentralGoogle Scholar
- 44.Gustin A, Kirchmeyer M, Koncina E, Felten P, Losciuto S, Heurtaux T, Tardivel A, Heuschling P, Dostert C (2015) NLRP3 inflammasome is expressed and functional in mouse brain microglia but not in astrocytes. PloS ONE 10(6):e0130624. https://doi.org/10.1371/journal.pone.0130624 CrossRefPubMedPubMedCentralGoogle Scholar
- 48.Minutoli L, Puzzolo D, Rinaldi M, Irrera N, Marini H, Arcoraci V, Bitto A, Crea G, Pisani A, Squadrito F, Trichilo V (2016) ROS-mediated NLRP3 inflammasome activation in brain, heart, kidney, and testis ischemia/reperfusion injury. Oxid Med Cell Longev. https://doi.org/10.1155/2016/2183026 PubMedPubMedCentralGoogle Scholar