P2X7 signaling promotes microsphere embolism-triggered microglia activation by maintaining elevation of Fas ligand

Journal of Neuroinflammation

Feng Han³  Ze-Hui Hong²  Kohji Fukunaga¹  Xiao-ming Li⁴  Mei-hua Liao³  Li-tao Li³  Ji-yun Huang³  Rong-rong Tao³  Ying-mei Lu⁴ 

[1] Department of Pharmacology, Graduate School of Pharmaceutical Sciences, Tohoku University, Sendai, Japan;The Key Laboratory of Developmental Genes and Human Disease, Institute of Life Science, Southeast University, Nanjing, China;Institute of Pharmacology, Toxicology and Biochemical Pharmaceutics, Zhejiang University, 866 Yu-Hang-Tang Road, Hangzhou, 310058, China;Department of Neurobiology, Key Laboratory of Medical Neurobiology of Ministry of Health of China, Zhejiang University School of Medicine, Hangzhou, China

关键词: Neuronal cell death;    Fas ligand;    P2X₇ receptor;    Microglia;    Microsphere embolism;   

Others  :  1160377
DOI  :  10.1186/1742-2094-9-172

 received in 2012-04-02, accepted in 2012-06-20,  发布年份 2012

【 摘 要 】

Background

The cerebral microvascular occlusion elicits microvascular injury which mimics the different degrees of stroke severity observed in patients, but the mechanisms underlying these embolic injuries are far from understood. The Fas ligand (FasL)-Fas system has been implicated in a number of pathogenic states. Here, we examined the contribution of microglia-derived FasL to brain inflammatory injury, with a focus on the potential to suppress the FasL increase by inhibition of the P2X₇-FasL signaling with pharmacological or genetic approaches during ischemia.

Methods

The cerebral microvascular occlusion was induced by microsphere injection in experimental animals. Morphological changes in microglial cells were studied immunohistochemically. The biochemical analyses were used to examine the intracellular changes of P2X₇/FasL signaling. The BV-2 cells and primary microglia from mice genetically deficient in P2X₇ were used to further establish a linkage between microglia activation and FasL overproduction.

Results

The FasL expression was continuously elevated and was spatiotemporally related to microglia activation following microsphere embolism. Notably, P2X₇ expression concomitantly increased in microglia and presented a distribution pattern that was similar to that of FasL in ED1-positive cells at pathological process of microsphere embolism. Interestingly, FasL generation in cultured microglia cells subjected to oxygen-glucose deprivation-treated neuron-conditioned medium was prevented by the silencing of P2X₇. Furthermore, FasL induced the migration of BV-2 microglia, whereas the neutralization of FasL with a blocking antibody was highly effective in inhibiting ischemia-induced microglial mobility. Similar results were observed in primary microglia from wild-type mice or mice genetically deficient in P2X₇. Finally, the degrees of FasL overproduction and neuronal death were consistently reduced in P2X₇^−/− mice compared with wild-type littermates following microsphere embolism insult.

Conclusion

FasL functions as a key component of an immunoreactive response loop by recruiting microglia to the lesion sites through a P2X₇-dependent mechanism. The specific modulation of P2X₇/FasL signaling and aberrant microglial activation could provide therapeutic benefits in acute and subacute phase of cerebral microembolic injury.

【 授权许可】

   
2012 han et al.

【 图 表 】

Figure 2.

Figure 3.

Figure 1.

Figure 5.

Figure 6.

Figure 7.

Figure 8.

Figure 9.

Figure 10.

【 参考文献 】

• [1]del Zoppo GJ: Microvascular changes during cerebral ischemia and reperfusion. Cerebrovasc Brain Metab Rev 1994, 6:47-96.
• [2]Lam CK, Yoo T, Hiner B, Liu Z, Grutzendler J: Embolus extravasation is an alternative mechanism for cerebral microvascular recanalization. Nature 2010, 465:478-482.
• [3]Iadecola C, Anrather J: The immunology of stroke: from mechanisms to translation. Nat Med 2011, 17:796-808.
• [4]Han F, Shioda N, Moriguchi S, Qin ZH, Fukunaga K: Downregulation of glutamate transporters is associated with elevation in extracellular glutamate concentration following rat microsphere embolism. Neurosci Lett 2008, 430:275-280.
• [5]Han F, Shirasaki Y, Fukunaga K: Microsphere embolism-induced endothelial nitric oxide synthase expression mediates disruption of the blood–brain barrier in rat brain. J Neurochem 2006, 99:97-106.
• [6]Han F, Tao RR, Zhang GS, Lu YM, Liu LL, Chen YX, Lou YJ, Fukunaga K, Hong ZH: Melatonin ameliorates ischaemic-like injury-evoked nitrosative stress: Involvement of HtrA2/PED pathways in endothelial cells. J Pineal Res 2011, 50:281-291.
• [7]del Zoppo GJ, Milner R, Mabuchi T, Hung S, Wang X, Berg GI, Koziol JA: Microglial activation and matrix protease generation during focal cerebral ischemia. Stroke 2007, 38(Suppl 2):646-651.
• [8]Macrez R, Ali C, Toutirais O, Le Mauff B, Defer G, Dirnagl U, Vivien D: Stroke and the immune system: from pathophysiology to new therapeutic strategies. Lancet Neurol 2011, 10:471-480.
• [9]Masuda T, Croom D, Hida H, Kirov SA: Capillary blood flow around microglial somata determines dynamics of microglial processes in ischaemic conditions. Glia 2011, 59:1744-1753.
• [10]Lambert C, Landau AM, Desbarats J: Fas-beyond death: a regenerative role for Fas in the nervous system. Apoptosis 2003, 8:551-562.
• [11]Fukunaga K, Ishigami T, Kawano T: Transcriptional regulation of neuronal genes and its effect on neural functions: expression and function of forkhead transcription factors in neurons. J Pharmacol Sci 2005, 98:205-211.
• [12]Martin-Villalba A, Hahne M, Kleber S, Vogel J, Falk W, Schenkel J, Krammer PH: Therapeutic neutralization of CD95-ligand and TNF attenuates brain damage in stroke. Cell Death Differ 2001, 8:679-686.
• [13]Choi C, Benveniste EN: Fas ligand/Fas system in the brain: regulator of immune and apoptotic responses. Brain Res Brain Res Rev 2004, 44:65-81.
• [14]Apenberg S, Freyberg MA, Friedl P: Shear stress induces apoptosis in vascular smooth muscle cells via an autocrine Fas/FasL pathway. Biochem Biophys Res Commun 2003, 310:355-359.
• [15]Tan KH, Hunziker W: Compartmentalization of Fas and Fas ligand may prevent auto- or paracrine apoptosis in epithelial cells. Exp Cell Res 2003, 284:283-290.
• [16]Hallermalm K, De Geer A, Kiessling R, Levitsky V, Levitskaya J: Autocrine secretion of Fas ligand shields tumor cells from Fas-mediated killing by cytotoxic lymphocytes. Cancer Res 2004, 64:6775-6782.
• [17]Giovannini C, Matarrese P, Scazzocchio B, Vari R, D’Archivio M, Straface E, Masella R, Malorni W, De Vicenzi M: Wheat gliadin induces apoptosis of intestinal cells via an autocrine mechanism involving Fas-Fas ligand pathway. FEBS Lett 2003, 540:117-124.
• [18]Iadecola C, Sugimoto K, Niwa K, Kazama K, Ross ME: Increased susceptibility to ischaemic brain injury in cyclooxygenase-1-deficient mice. J Cereb Blood Flow Metab 2001, 21:1436-1441.
• [19]Jin G, Hayashi T, Kawagoe J, Takizawa T, Nagata T, Nagano I, Syoji M, Abe K: Deficiency of PAR-2 gene increases acute focal ischaemic brain injury. J Cereb Blood Flow Metab 2005, 25:302-313.
• [20]Kawano T, Anrather J, Zhou P, Park L, Wang G, Frys KA, Kunz A, Cho S, Orio M, Iadecola C: Prostaglandin E2 EP1 receptors: downstream effectors of COX-2 neurotoxicity. Nat Med 2006, 12:225-229.
• [21]Yenari MA, Xu L, Tang XN, Qiao Y, Giffard RG: Microglia potentiate damage to blood–brain barrier constituents: improvement by minocycline in vivo and in vitro. Stroke 2006, 37:1087-1093.
• [22]Orio M, Kunz A, Kawano T, Anrather J, Zhou P, Iadecola C: Lipopolysaccharide induces early tolerance to excitotoxicity via nitric oxide and cGMP. Stroke 2007, 38:2812-2817.
• [23]Vogt M, Bauer MK, Ferrari D, Schulze-Osthoff K: Oxidative stress and hypoxia/reoxygenation trigger CD95 (APO-1/Fas) ligand expression in microglial cells. FEBS Lett 1998, 429:67-72.
• [24]Facchinetti F, Furegato S, Terrazzino S, Leon A: H(2)O(2) induces upregulation of Fas and Fas ligand expression in NGF-differentiated PC12 cells: modulation by cAMP. J Neurosci Res 2002, 69:178-188.
• [25]Ekdahl CT, Kokaia Z, Lindvall O: Brain inflammation and adult neurogenesis: the dual role of microglia. Neuroscience 2009, 158:1021-1029.
• [26]Gao F, Wang S, Guo Y, Wang J, Lou M, Wu J, Ding M, Tian M, Zhang H: Protective effects of repetitive transcranial magnetic stimulation in a rat model of transient cerebral ischaemia: a microPET study. Eur J Nucl Med Mol Imaging 2010, 37:954-961.
• [27]Kim TK, Hemberg M, Gray JM, Costa AM, Bear DM, Wu J, Harmin DA, Laptewicz M, Barbara-Haley K, Kuersten S, Markenscoff-Papadimitriou E, Kuhl D, Bito H, Worley PF, Kreiman G, Greenberg ME: Widespread transcription at neuronal activity-regulated enhancers. Nature 2010, 465:182-187.
• [28]Ullrich O, Diestel A, Bechmann I, Homberg M, Grune T, Hass R, Nitsch R: Turnover of oxidatively damaged nuclear proteins in BV-2 microglial cells is linked to their activation state by poly-ADP-ribose polymerase. FASEB J 2001, 15:1460-1462.
• [29]Jang S, Kelley KW, Johnson RW: Luteolin reduces IL-6 production in microglia by inhibiting JNK phosphorylation and activation of AP-1. Proc Natl Acad Sci USA 2008, 105:7534-7539.
• [30]Honda S, Sasaki Y, Ohsawa K, Imai Y, Nakamura Y, Inoue K, Kohsaka S: Extracellular ATP or ADP induce chemotaxis of cultured microglia through Gi/o-coupled P2Y receptors. J Neurosci 2001, 21:1975-1982.
• [31]Giulian D, Baker TJ: Characterization of ameboid microglia isolated from developing mammalian brain. J Neurosci 1986, 6:2163-2178.
• [32]Dou Y, Wu HJ, Li HQ, Qin S, Wang YE, Li J, Lou HF, Chen Z, Li XM, Luo QM, Duan S: Microglial migration mediated by ATP-induced ATP release from lysosomes. Cell Res 2012, 22:1022-1033.
• [33]Schmued LC, Stowers CC, Scallet AC, Xu L: Fluoro-Jade C results in ultra high resolution and contrast labeling of degenerating neurons. Brain Res 2005, 1035:24-31.
• [34]Glezer I, Simard AR, Rivest S: Neuroprotective role of the innate immune system by microglia. Neuroscience 2007, 147:867-883.
• [35]Ito D, Imai Y, Ohsawa K, Nakajima K, Fukuuchi Y, Kohsaka S: Microglia-specific localisation of a novel calcium binding protein, Iba1. Brain Res Mol Brain Res 1998, 57:1-9.
• [36]Witting A, Walter L, Wacker J, Möller T, Stella N: P2X7 receptors control 2-arachidonoylglycerol production by microglial cells. Proc Natl Acad Sci USA 2004, 101:3214-3219.
• [37]Sharp AJ, Polak PE, Simonini V, Lin SX, Richardson JC, Bongarzone ER, Feinstein DL: P2x7 deficiency suppresses development of experimental autoimmune encephalomyelitis. J Neuroinflammation 2008, 5:33. BioMed Central Full Text
• [38]Kim JE, Ryu HJ, Yeo SI, Kang TC: P2X7 receptor regulates leukocyte infiltrations in rat frontoparietal cortex following status epilepticus. J Neuroinflammation 2010, 7:65. BioMed Central Full Text
• [39]Melani A, Amadio S, Gianfriddo M, Vannucchi MG, Volontè C, Bernardi G, Pedata F, Sancesario G: P2X7 receptor modulation on microglial cells and reduction of brain infarct caused by middle cerebral artery occlusion in rat. J Cereb Blood Flow Metab 2006, 26:974-982.
• [40]Tang M, Alexander H, Clark RS, Kochanek PM, Kagan VE, Bayir H: Minocycline reduces neuronal death and attenuates microglial response after pediatric asphyxial cardiac arrest. J Cereb Blood Flow Metab 2010, 30:119-129.
• [41]Henry CJ, Huang Y, Wynne A, Hanke M, Himler J, Bailey MT, Sheridan JF, Godbout JP: Minocycline attenuates lipopolysaccharide (LPS)-induced neuroinflammation, sickness behavior, and anhedonia. J Neuroinflammation 2008, 5:15. BioMed Central Full Text
• [42]Streit WJ, Mrak RE, Griffin WS: Microglia and neuroinflammation: a pathological perspective. J Neuroinflammation 2004, 1:14. BioMed Central Full Text
• [43]Griffin WS: Inflammation and neurodegenerative diseases. Am J Clin Nutr 2006, 83:470S-474S.
• [44]Mrak RE, Griffin WS: Glia and their cytokines in progression of neurodegeneration. Neurobiol Aging 2005, 26:349-354.
• [45]Davalos D, Grutzendler J, Yang G, Kim JV, Zuo Y, Jung S, Littman DR, Dustin ML, Gan WB: ATP mediates rapid microglial response to local brain injury in vivo. Nat Neurosci 2005, 8:752-758.
• [46]Bechmann I, Mor G, Nilsen J, Eliza M, Nitsch R, Naftolin F: FasL (CD95L, Apo1L) is expressed in the normal rat and human brain: evidence for the existence of an immunological brain barrier. Glia 1999, 27:62-74.
• [47]Beer R, Franz G, Schöpf M, Reindl M, Zelger B, Schmutzhard E, Poewe W, Kampfl A: Expression of Fas and Fas ligand after experimental traumatic brain injury in the rat. J Cereb Blood Flow Metab 2000, 20:669-677.
• [48]Terrazzino S, Bauleo A, Baldan A, Leon A: Peripheral LPS administrations up-regulate Fas and FasL on brain microglial cells: a brain protective or pathogenic event? J Neuroimmunol 2002, 124:45-53.
• [49]Clark AK, Wodarski R, Guida F, Sasso O, Malcangio M: Cathepsin S release from primary cultured microglia is regulated by the P2X7 receptor. Glia 2010, 58:1710-1726.
• [50]Monif M, Reid CA, Powell KL, Smart ML, Williams DA: The P2X7 receptor drives microglial activation and proliferation: a trophic role for P2X7R pore. J Neurosci 2009, 29:3781-3791.
• [51]Chen KC, Liu WH, Chang LS: Suppression of ERK signaling evokes autocrine Fas-mediated death in arachidonic acid-treated human chronic myeloid leukemia K562 cells. J Cell Physiol 2010, 222:625-634.
• [52]Becher B, Dodelet V, Fedorowicz V, Antel JP: Soluble tumor necrosis factor receptor inhibits interleukin 12 production by stimulated human adult microglial cells in vitro. J Clin Invest 1996, 98:1539-1543.
• [53]Bezzi P, Domercq M, Brambilla L, Galli R, Schols D, De Clercq E, Vescovi A, Bagetta G, Kollias G, Meldolesi J, Volterra A: CXCR4-activated astrocyte glutamate release via TNFalpha: amplification by microglia triggers neurotoxicity. Nat Neurosci 2001, 4:702-710.
• [54]Meucci O, Fatatis A, Simen AA, Miller RJ: Expression of CX3CR1 chemokine receptors on neurons and their role in neuronal survival. Proc Natl Acad Sci USA 2000, 97:8075-8080.
• [55]Ramsdell F, Seaman MS, Miller RE, Tough TW, Alderson MR, Lynch DH: gld/gld mice are unable to express a functional ligand for Fas. Eur J Immunol 1994, 24:928-933.
• [56]O’Connell J, Houston A, Bennett MW, O’Sullivan GC, Shanahan F: Immune privilege or inflammation? Insights into the Fas ligand enigma. Nat Med 2001, 7:271-274.
• [57]Kim JV, Dustin ML: Innate response to focal necrotic injury inside the blood–brain barrier. J Immunol 2006, 177:5269-5277.
• [58]Saijo K, Winner B, Carson CT, Collier JG, Boyer L, Rosenfeld MG, Gage FH, Glass CK: A Nurr1/CoREST pathway in microglia and astrocytes protects dopaminergic neurons from inflammation-induced death. Cell 2009, 137:47-59.
• [59]Bonde S, Ekdahl CT, Lindvall O: Long-term neuronal replacement in adult rat hippocampus after status epilepticus despite chronic inflammation. Eur J Neurosci 2006, 23:965-974.