Abstract
Complement-mediated inflammation plays a vital role in intracerebral hemorrhage (ICH), implicating pro-inflammatory factor interleukin-1beta (IL-1β) secretion. Brain samples and contralateral hemiencephalon were all collected and detected by Western blot. NLRP3 expression was located by dual immunofluorescence staining at 1, 3, and 5 days post-ICH. Brain water content was examined post-ICH. The neural deficit scores were evaluated by observers blindly. ILs were detected by ELISA. SiRNAs targeting NLRP3 (siNLRP3), siASC, and siControl were injected to inhibit NLRP3 function. To test the complement activation via Nod-like receptor (NLR) family pyrin domain-containing 3 (NLRP3), normal rabbit complement (NRC) was injected with lipopolysaccharide (LPS) to facilitate the complement function. As a result, complement 3a (C3a) and complement 5a (C5a) were upregulated during the ICH-induced neuroinflammation, and ablation of C3 attenuates ICH-induced IL-1β release. Though the LPS rescues the neuroinflammation in the ICH model, C3 deficiency attenuates the LPS-induced inflammatory effect. The NLRP3 inflammasome was activated after ICH and was located in the microglial cell of the mouse brain, which exhibits a time-dependent manner. However, the number of NLRP3/Iba-1 dual-labeled cells in the C3−/− group is less than that in the WT group in each time course, respectively. IL-1β and IL-18 released in perihematoma tissue, caspase-1-p20, brain water content, and behavioral outcomes were attenuated in the siNLRP3 and siASC groups than in the siControl and ICH groups. We also found that 5% of complement supplement enhances ICH-induced IL-1β release, while NLRP3 and ASC inhibition attenuates it. In conclusion, complement-induced ICH neuroinflammation depended on NLRP3 activation, which facilities LPS- and ICH-induced neuroinflammation, and NLRP3 is required for ICH-induced inflammation.
Similar content being viewed by others
References
Block ML, Zecca L, Hong JS (2007) Microglia-mediated neurotoxicity: uncovering the molecular mechanisms. Nat Rev Neurosci 8:57–69. doi:10.1038/nrn2038
Bonnardeaux A, Pichette V (2003) Complement dysregulation in haemolytic uraemic syndrome. Lancet 362:1514–1515. doi:10.1016/S0140-6736(03)14777-0
Chen GY, Nunez G (2010) Sterile inflammation: sensing and reacting to damage. Nat Rev Immunol 10:826–837. doi:10.1038/nri2873
Couto Alves A et al (2013) Dysregulation of complement system and CD4+ T cell activation pathways implicated in allergic response. PLoS One 8:e74821. doi:10.1371/journal.pone.0074821
Dinarello CA (2009) Immunological and inflammatory functions of the interleukin-1 family. Annu Rev Immunol 27:519–550. doi:10.1146/annurev.immunol.021908.132612
Dinarello CA (2011) Interleukin-1 in the pathogenesis and treatment of inflammatory diseases. Blood 117:3720–3732. doi:10.1182/blood-2010-07-273417
Dostert C, Petrilli V, Van Bruggen R, Steele C, Mossman BT, Tschopp J (2008) Innate immune activation through Nalp3 inflammasome sensing of asbestos and silica. Science 320:674–677. doi:10.1126/science.1156995
Doyle SL et al (2012) NLRP3 has a protective role in age-related macular degeneration through the induction of IL-18 by drusen components. Nat Med 18:791–798. doi:10.1038/nm.2717
Duewell P et al (2010) NLRP3 inflammasomes are required for atherogenesis and activated by cholesterol crystals. Nature 464:1357–1361. doi:10.1038/nature08938
Fang C, Zhang X, Miwa T, Song WC (2009) Complement promotes the development of inflammatory T-helper 17 cells through synergistic interaction with Toll-like receptor signaling and interleukin-6 production. Blood 114:1005–1015. doi:10.1182/blood-2009-01-198283
Fann DY et al (2013) Intravenous immunoglobulin suppresses NLRP1 and NLRP3 inflammasome-mediated neuronal death in ischemic stroke. Cell Death Dis 4:e790. doi:10.1038/cddis.2013.326
Flower O, Smith M (2011) The acute management of intracerebral hemorrhage. Curr Opin Crit Care 17:106–114. doi:10.1097/MCC.0b013e328342f823
Fogal B, Li J, Lobner D, McCullough LD, Hewett SJ (2007) System x(c)- activity and astrocytes are necessary for interleukin-1 beta-mediated hypoxic neuronal injury. The Journal of neuroscience : the official journal of the Society for Neuroscience 27:10094–10105. doi:10.1523/JNEUROSCI.2459-07.2007
Forte A, Cipollaro M, Cascino A, Galderisi U (2005) Small interfering RNAs and antisense oligonucleotides for treatment of neurological diseases. Curr Drug Targets 6:21–29
Fountaine TM, Wood MJ, Wade-Martins R (2005) Delivering RNA interference to the mammalian brain. Current gene therapy 5:399–410
Fure H, Nielsen EW, Hack CE, Mollnes TE (1997) A neoepitope-based enzyme immunoassay for quantification of C1-inhibitor in complex with C1r and C1s. Scand J Immunol 46:553–557
Garrett MC et al (2009) Synergistic neuroprotective effects of C3a and C5a receptor blockade following intracerebral hemorrhage. Brain Res 1298:171–177. doi:10.1016/j.brainres.2009.04.047
Goldmann T, Tay TL, Prinz M (2013) Love and death: microglia, NLRP3 and the Alzheimer’s brain. Cell Res 23:595–596. doi:10.1038/cr.2013.24
Goldstein JN, Gilson AJ (2011) Critical care management of acute intracerebral hemorrhage. Curr Treat Options Neurol 13:204–216. doi:10.1007/s11940-010-0109-2
Halle A et al (2008) The NALP3 inflammasome is involved in the innate immune response to amyloid-beta. Nat Immunol 9:857–865. doi:10.1038/ni.1636
Hanamsagar R, Hanke ML, Kielian T (2012) Toll-like receptor (TLR) and inflammasome actions in the central nervous system. Trends Immunol 33:333–342. doi:10.1016/j.it.2012.03.001
Heeringa SF, Cohen CD (2012) Kidney diseases caused by complement dysregulation: acquired, inherited, and still more to come. Clinical & developmental immunology 2012:695131. doi:10.1155/2012/695131
Hoegen T et al (2011) The NLRP3 inflammasome contributes to brain injury in pneumococcal meningitis and is activated through ATP-dependent lysosomal cathepsin B release. J Immunol 187:5440–5451. doi:10.4049/jimmunol.1100790
Hwang BY et al (2011) Advances in neuroprotective strategies: potential therapies for intracerebral hemorrhage. Cerebrovasc Dis 31:211–222. doi:10.1159/000321870
Jiang Y, Ma J, Li H, Liu Y, You C (2015) Effect of apolipoprotein C3 genetic polymorphisms on serum lipid levels and the risk of intracerebral hemorrhage. Lipids Health Dis 14:48. doi:10.1186/s12944-015-0047-9
Laudisi F et al (2013) Cutting edge: the NLRP3 inflammasome links complement-mediated inflammation and IL-1beta release. J Immunol 191:1006–1010. doi:10.4049/jimmunol.1300489
Li G et al (2013) Neuroprotective effects of Argatroban and C5a receptor antagonist (PMX53) following intracerebral hemorrhage. Clin Exp Immunol. doi:10.1111/cei.12220
Lin S et al (2012) Heme activates TLR4-mediated inflammatory injury via MyD88/TRIF signaling pathway in intracerebral hemorrhage. J Neuroinflammation 9:46. doi:10.1186/1742-2094-9-46
Lint TF, Behrends CL, Baker PJ, Gewurz H (1976) Activation of the complement attack mechanism in the fluid phase and its control by C567-INH: lysis of normal erythrocytes initiated by zymosan, endotoxin, and immune complexes. J Immunol 117:1440–1446
Lok J et al (2012) Neuregulin-1 effects on endothelial and blood-brain-barrier permeability after experimental injury. Translational stroke research 3(Suppl 1):S119–S124. doi:10.1007/s12975-012-0157-x
Lu A, Tang Y, Ran R, Ardizzone TL, Wagner KR, Sharp FR (2006) Brain genomics of intracerebral hemorrhage. Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism 26:230–252. doi:10.1038/sj.jcbfm.9600183
Mariathasan S, Monack DM (2007) Inflammasome adaptors and sensors: intracellular regulators of infection and inflammation. Nat Rev Immunol 7:31–40. doi:10.1038/nri1997
Martinon F, Petrilli V, Mayor A, Tardivel A, Tschopp J (2006) Gout-associated uric acid crystals activate the NALP3 inflammasome. Nature 440:237–241. doi:10.1038/nature04516
Masada T, Hua Y, Xi G, Yang GY, Hoff JT, Keep RF (2001) Attenuation of intracerebral hemorrhage and thrombin-induced brain edema by overexpression of interleukin-1 receptor antagonist. J Neurosurg 95:680–686. doi:10.3171/jns.2001.95.4.0680
Mollnes TE et al (2002) Essential role of the C5a receptor in E coli-induced oxidative burst and phagocytosis revealed by a novel lepirudin-based human whole blood model of inflammation. Blood 100:1869–1877
Nakamura T, Xi G, Hua Y, Schallert T, Hoff JT, Keep RF (2004) Intracerebral hemorrhage in mice: model characterization and application for genetically modified mice. Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism 24:487–494. doi:10.1097/00004647-200405000-00002
Petrilli V, Dostert C, Muruve DA, Tschopp J (2007) The inflammasome: a danger sensing complex triggering innate immunity. Curr Opin Immunol 19:615–622. doi:10.1016/j.coi.2007.09.002
Qureshi AI, Mendelow AD, Hanley DF (2009) Intracerebral haemorrhage. Lancet 373:1632–1644. doi:10.1016/S0140-6736(09)60371-8
Rathinam VA, Vanaja SK, Fitzgerald KA (2012) Regulation of inflammasome signaling. Nat Immunol 13:333–342. doi:10.1038/ni.2237
Ribo M, Grotta JC (2006) Latest advances in intracerebral hemorrhage. Current neurology and neuroscience reports 6:17–22
Rynkowski MA et al (2009) C3a receptor antagonist attenuates brain injury after intracerebral hemorrhage. Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism 29:98–107. doi:10.1038/jcbfm.2008.95
Samstad EO et al (2014) Cholesterol crystals induce complement-dependent inflammasome activation and cytokine release. J Immunol 192:2837–2845. doi:10.4049/jimmunol.1302484
Sansing LH, Kaznatcheeva EA, Perkins CJ, Komaroff E, Gutman FB, Newman GC (2003) Edema after intracerebral hemorrhage: correlations with coagulation parameters and treatment. J Neurosurg 98:985–992. doi:10.3171/jns.2003.98.5.0985
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 3:288. doi:10.3389/fimmu.2012.00288
Srinivasan D, Yen JH, Joseph DJ, Friedman W (2004) Cell type-specific interleukin-1beta signaling in the CNS. The Journal of neuroscience : the official journal of the Society for Neuroscience 24:6482–6488. doi:10.1523/JNEUROSCI.5712-03.2004
Stahel PF et al (2000) Intracerebral complement C5a receptor (CD88) expression is regulated by TNF and lymphotoxin-alpha following closed head injury in mice. J Neuroimmunol 109:164–172
Strowig T, Henao-Mejia J, Elinav E, Flavell R (2012) Inflammasomes in health and disease. Nature 481:278–286. doi:10.1038/nature10759
Stutz A, Golenbock DT, Latz E (2009) Inflammasomes: too big to miss. J Clin Invest 119:3502–3511. doi:10.1172/JCI40599
Sukumari-Ramesh S, Alleyne CH Jr (2016) Post-injury administration of tert-butylhydroquinone attenuates acute neurological injury after intracerebral hemorrhage in mice. Journal of molecular neuroscience : MN 58:525–531. doi:10.1007/s12031-016-0722-y
Suresh R, Chandrasekaran P, Sutterwala FS, Mosser DM (2016) Complement-mediated ‘bystander’ damage initiates host NLRP3 inflammasome activation. J Cell Sci 129:1928–1939. doi:10.1242/jcs.179291
Sutterwala FS, Haasken S, Cassel SL (2014) Mechanism of NLRP3 inflammasome activation. Ann N Y Acad Sci 1319:82–95. doi:10.1111/nyas.12458
van Asch CJ, Luitse MJ, Rinkel GJ, van der Tweel I, Algra A, Klijn CJ (2010) Incidence, case fatality, and functional outcome of intracerebral haemorrhage over time, according to age, sex, and ethnic origin: a systematic review and meta-analysis. Lancet Neurol 9:167–176. doi:10.1016/S1474-4422(09)70340-0
Wei P, You C, Jin H, Chen H, Lin B (2014) Correlation between serum IL-1beta levels and cerebral edema extent in a hypertensive intracerebral hemorrhage rat model. Neurol Res 36:170–175. doi:10.1179/1743132813Y.0000000292
Wu B, Ma Q, Khatibi N, Chen W, Sozen T, Cheng O, Tang J (2010) Ac-YVAD-CMK decreases blood-brain barrier degradation by inhibiting caspase-1 activation of interleukin-1beta in intracerebral hemorrhage mouse model. Translational stroke research 1:57–64. doi:10.1007/s12975-009-0002-z
Xi G, Keep RF, Hoff JT (2002) Pathophysiology of brain edema formation. Neurosurg Clin N Am 13:371–383
Yang S et al (2006) The role of complement C3 in intracerebral hemorrhage-induced brain injury. Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism 26:1490–1495. doi:10.1038/sj.jcbfm.9600305
Yang F et al (2014) NLRP3 deficiency ameliorates neurovascular damage in experimental ischemic stroke. Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism. doi:10.1038/jcbfm.2013.242
Yuen CM, Chiu CA, Chang LT, Liou CW, Lu CH, Youssef AA, Yip HK (2007) Level and value of interleukin-18 after acute ischemic stroke. Circulation journal : official journal of the Japanese Circulation Society 71:1691–1696
Zhang X et al (2007) Regulation of Toll-like receptor-mediated inflammatory response by complement in vivo. Blood 110:228–236. doi:10.1182/blood-2006-12-063636
Acknowledgements
This work was supported by the National Natural Science Foundation of China (81660211) and the Major Program of Science and Technology Foundation of Zunyi (Guizhou) Technology Bureau.
Author Contributions
STY was involved in performing the majority of the laboratory-based work and writing of the manuscript. JLC, WC, and RMF helped with laboratory work concerning control tissues and were involved in the writing of the manuscript. GL, YCZ, SJ, XPX, and CH were involved in collecting data, statistics, and troubleshooting. STY and PW were involved in conceptualizing and planning of the presented work.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
All animal studies followed the guidelines outlined in the Guide for the Care and Use of Laboratory Animals from the National Institute of Health and were approved by the Zunyi Hospital of Zunyi Animal Welfare Committee.
Conflict of Interest
The authors declare that they have no conflict of interest.
Additional information
Sheng-Tao Yao and Fang Cao contributed equally to the manuscript.
Rights and permissions
About this article
Cite this article
Yao, ST., Cao, F., Chen, JL. et al. NLRP3 is Required for Complement-Mediated Caspase-1 and IL-1beta Activation in ICH. J Mol Neurosci 61, 385–395 (2017). https://doi.org/10.1007/s12031-016-0874-9
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12031-016-0874-9