Neuronal CCL2 is upregulated during hepatic encephalopathy and contributes to microglia activation and neurological decline

Journal of Neuroinflammation

Sharon DeMorrow²  Gianfranco Alpini²  Eric Whittington³  Holly Standeford²  Cheryl Galindo²  Michelle Thompson¹  Gabriel Frampton²  Matthew McMillin² 

[1] Digestive Disease Research Center, Scott & White Hospital, 1901 South 1st Street, Building 205, Temple, Texas, USA;Central Texas Veterans Healthcare System, 1901 South 1st Street, Building 205, Temple, Texas, USA;Department of Internal Medicine, Texas A&M Health Science Center, College of Medicine, 1901 South 1st Street, Building 205, Temple, Texas, USA

关键词: Neuroinflammation;    CCR4;    CCR2;    Azoxymethane;    Acute liver failure;   

Others  :  1151345
DOI  :  10.1186/1742-2094-11-121

 received in 2014-02-03, accepted in 2014-06-30,  发布年份 2014

【 摘 要 】

Background

Acute liver failure leads to systemic complications with one of the most dangerous being a decline in neurological function, termed hepatic encephalopathy. Neurological dysfunction is exacerbated by an increase of toxic metabolites in the brain that lead to neuroinflammation. Following various liver diseases, hepatic and circulating chemokines, such as chemokine ligand 2 (CCL2), are elevated, though their effects on the brain following acute liver injury and subsequent hepatic encephalopathy are unknown. CCL2 is known to activate microglia in other neuropathies, leading to a proinflammatory response. However, the effects of CCL2 on microglia activation and the pathogenesis of hepatic encephalopathy following acute liver injury remain to be determined.

Methods

Hepatic encephalopathy was induced in mice via injection of azoxymethane (AOM) in the presence or absence of INCB 3284 dimesylate (INCB), a chemokine receptor 2 inhibitor, or C 021 dihydrochloride (C021), a chemokine receptor 4 inhibitor. Mice were monitored for neurological decline and time to coma (loss of all reflexes) was recorded. Tissue was collected at coma and used for real-time PCR, immunoblots, ELISA, or immunostaining analyses to assess the activation of microglia and consequences on pro-inflammatory cytokine expression.

Results

Following AOM administration, microglia activation was significantly increased in AOM-treated mice compared to controls. Concentrations of CCL2 in the liver, serum, and cortex were significantly elevated in AOM-treated mice compared to controls. Systemic administration of INCB or C021 reduced liver damage as assessed by serum liver enzyme biochemistry. Administration of INCB or C021 significantly improved the neurological outcomes of AOM-treated mice, reduced microglia activation, reduced phosphorylation of ERK1/2, and alleviated AOM-induced cytokine upregulation.

Conclusions

These findings suggest that CCL2 is elevated systemically following acute liver injury and that CCL2 is involved in both the microglia activation and neurological decline associated with hepatic encephalopathy. Methods used to modulate CCL2 levels and/or reduce CCR2/CCR4 activity may be potential therapeutic targets for the management of hepatic encephalopathy due to acute liver injury.

【 授权许可】

   
2014 McMillin et al.; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Butterworth RF: Hepatic encephalopathy: a central neuroinflammatory disorder? Hepatology 2011, 53:1372-1376.
• [2]Hazell AS, Butterworth RF: Hepatic encephalopathy: an update of pathophysiologic mechanisms. Proc Soc Exp Biol Med 1999, 222:99-112.
• [3]Butterworth RF: Complications of cirrhosis III. Hepatic encephalopathy. J Hepatol 2000, 32:171-180.
• [4]Fridman V, Galetta SL, Pruitt AA, Levine JM: MRI findings associated with acute liver failure. Neurology 2009, 72:2130-2131.
• [5]Wright G, Shawcross D, Olde Damink SW, Jalan R: Brain cytokine flux in acute liver failure and its relationship with intracranial hypertension. Metab Brain Dis 2007, 22:375-388.
• [6]Bemeur C, Qu H, Desjardins P, Butterworth RF: IL-1 or TNF receptor gene deletion delays onset of encephalopathy and attenuates brain edema in experimental acute liver failure. Neurochem Int 2010, 56:213-215.
• [7]Jiang W, Desjardins P, Butterworth RF: Cerebral inflammation contributes to encephalopathy and brain edema in acute liver failure: protective effect of minocycline. J Neurochem 2009, 109:485-493.
• [8]Semple BD, Kossmann T, Morganti-Kossmann MC: Role of chemokines in CNS health and pathology: a focus on the CCL2/CCR2 and CXCL8/CXCR2 networks. J Cereb Blood Flow Metab 2010, 30:459-473.
• [9]Che X, Ye W, Panga L, Wu DC, Yang GY: Monocyte chemoattractant protein-1 expressed in neurons and astrocytes during focal ischemia in mice. Brain Res 2001, 902:171-177.
• [10]Frampton G, Invernizzi P, Bernuzzi F, Pae HY, Quinn M, Horvat D, Galindo C, Huang L, McMillin M, Cooper B, Rimassa L, DeMorrow S: Interleukin-6-driven progranulin expression increases cholangiocarcinoma growth by an Akt-dependent mechanism. Gut 2012, 61:268-277.
• [11]DeMorrow S, Francis H, Gaudio E, Venter J, Franchitto A, Kopriva S, Onori P, Mancinelli R, Frampton G, Coufal M, Mitchell B, Vaculin B, Alpini G: The endocannabinoid anandamide inhibits cholangiocarcinoma growth via activation of the noncanonical Wnt signaling pathway. Am J Physiol Gastrointest Liver Physiol 2008, 295:G1150-G1158.
• [12]Livak KJ, Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 2001, 25:402-408.
• [13]DeMorrow S, Glaser S, Francis H, Venter J, Vaculin B, Vaculin S, Alpini G: Opposing actions of endocannabinoids on cholangiocarcinoma growth: Recruitment of fas and fas ligand to lipid rafts. J Biol Chem 2007, 282:13098-13113.
• [14]Jiang W, Desjardins P, Butterworth RF: Direct evidence for central proinflammatory mechanisms in rats with experimental acute liver failure: protective effect of hypothermia. J Cereb Blood Flow Metab 2009, 29:944-952.
• [15]Chuang SY, Yang SH, Pang JH: Cilostazol reduces MCP-1-induced chemotaxis and adhesion of THP-1 monocytes by inhibiting CCR2 gene expression. Biochem Biophys Res Commun 2011, 411:402-408.
• [16]D’Mello C, Le T, Swain MG: Cerebral microglia recruit monocytes into the brain in response to tumor necrosis factor alpha signaling during peripheral organ inflammation. J Neurosci 2009, 29:2089-2102.
• [17]Zemtsova I, Gorg B, Keitel V, Bidmon HJ, Schror K, Haussinger D: Microglia activation in hepatic encephalopathy in rats and humans. Hepatology 2011, 54:204-215.
• [18]Dennis CV, Sheahan PJ, Graeber MB, Sheedy DL, Kril JJ, Sutherland GT: Microglial proliferation in the brain of chronic alcoholics with hepatic encephalopathy. Metab Brain Dis 2013. In press
• [19]van Rossum D, Hanisch UK: Microglia. Metab Brain Dis 2004, 19:393-411.
• [20]Chastre A, Belanger M, Beauchesne E, Nguyen BN, Desjardins P, Butterworth RF: Inflammatory cascades driven by tumor necrosis factor-alpha play a major role in the progression of acute liver failure and its neurological complications. PLoS One 2012, 7:e49670.
• [21]Vaquero J, Belanger M, James L, Herrero R, Desjardins P, Cote J, Blei AT, Butterworth RF: Mild hypothermia attenuates liver injury and improves survival in mice with acetaminophen toxicity. Gastroenterology 2007, 132:372-383.
• [22]Rock RB, Hu S, Sheng WS, Peterson PK: Morphine stimulates CCL2 production by human neurons. J Neuroinflammation 2006, 3:32.
• [23]Conductier G, Blondeau N, Guyon A, Nahon JL, Rovere C: The role of monocyte chemoattractant protein MCP1/CCL2 in neuroinflammatory diseases. J Neuroimmunol 2010, 224:93-100.
• [24]Coughlan CM, McManus CM, Sharron M, Gao Z, Murphy D, Jaffer S, Choe W, Chen W, Hesselgesser J, Gaylord H, Kalyuzhny A, Lee VM, Wolf B, Doms RW, Kolson DL: Expression of multiple functional chemokine receptors and monocyte chemoattractant protein-1 in human neurons. Neuroscience 2000, 97:591-600.
• [25]Ait-Ali D, Almeida-Suhett CP, Li Z, Marini AM, Braga MF, Eiden LE: Temporal course of changes in gene expression suggests a cytokine-related mechanism for long-term hippocampal alteration after controlled cortical impact. J Neurotrauma 2014, 31(7):683-690.
• [26]Yao Y, Tsirka SE: The CCL2-CCR2 system affects the progression and clearance of intracerebral hemorrhage. Glia 2012, 60:908-918.
• [27]Arakelyan A, Petrkova J, Hermanova Z, Boyajyan A, Lukl J, Petrek M: Serum levels of the MCP-1 chemokine in patients with ischemic stroke and myocardial infarction. Mediators Inflamm 2005, 2005:175-179.
• [28]Corsi MM, Licastro F, Porcellini E, Dogliotti G, Galliera E, Lamont JL, Innocenzi PJ, Fitzgerald SP: Reduced plasma levels of P-selectin and L-selectin in a pilot study from Alzheimer disease: relationship with neuro-degeneration. Biogerontology 2011, 12:451-454.
• [29]Rhodes JK, Sharkey J, Andrews PJ: The temporal expression, cellular localization, and inhibition of the chemokines MIP-2 and MCP-1 after traumatic brain injury in the rat. J Neurotrauma 2009, 26:507-525.
• [30]Degre D, Lemmers A, Gustot T, Ouziel R, Trepo E, Demetter P, Verset L, Quertinmont E, Vercruysse V, Le Moine O, Devière J, Moreno C: Hepatic expression of CCL2 in alcoholic liver disease is associated with disease severity and neutrophil infiltrates. Clin Exp Immunol 2012, 169:302-310.
• [31]Bertola A, Bonnafous S, Anty R, Patouraux S, Saint-Paul MC, Iannelli A, Gugenheim J, Barr J, Mato JM, Le Marchand-Brustel Y, Tran A, Gual P: Hepatic expression patterns of inflammatory and immune response genes associated with obesity and NASH in morbidly obese patients. PLoS One 2010, 5:e13577.
• [32]Antoniades CG, Quaglia A, Taams LS, Mitry RR, Hussain M, Abeles R, Possamai LA, Bruce M, McPhail M, Starling C, Wagner B, Barnardo A, Pomplun S, Auzinger G, Bernal W, Heaton N, Vergani D, Thursz MR, Wendon J: Source and characterization of hepatic macrophages in acetaminophen-induced acute liver failure in humans. Hepatology 2012, 56:735-746.
• [33]Yang Q, Shi Y, He J, Chen Z: The evolving story of macrophages in acute liver failure. Immunol Lett 2012, 147:1-9.
• [34]Zamara E, Galastri S, Aleffi S, Petrai I, Aragno M, Mastrocola R, Novo E, Bertolani C, Milani S, Vizzutti F, Vercelli A, Pinzani M, Laffi G, LaVilla G, Parola M, Marra F: Prevention of severe toxic liver injury and oxidative stress in MCP-1-deficient mice. J Hepatol 2007, 46:230-238.
• [35]Egan CE, Daugherity EK, Rogers AB, Abi Abdallah DS, Denkers EY, Maurer KJ: CCR2 and CD44 promote inflammatory cell recruitment during fatty liver formation in a lithogenic diet fed mouse model. PLoS One 2013, 8:e65247.
• [36]Hogaboam CM, Bone-Larson CL, Steinhauser ML, Matsukawa A, Gosling J, Boring L, Charo IF, Simpson KJ, Lukacs NW, Kunkel SL: Exaggerated hepatic injury due to acetaminophen challenge in mice lacking C-C chemokine receptor 2. Am J Pathol 2000, 156:1245-1252.
• [37]Nguyen JH, Yamamoto S, Steers J, Sevlever D, Lin W, Shimojima N, Castanedes-Casey M, Genco P, Golde T, Richelson E, Dickson D, McKinney M, Eckman CB: Matrix metalloproteinase-9 contributes to brain extravasation and edema in fulminant hepatic failure mice. J Hepatol 2006, 44:1105-1114.
• [38]Strecker JK, Minnerup J, Schutte-Nutgen K, Gess B, Schabitz WR, Schilling M: Monocyte chemoattractant protein-1-deficiency results in altered blood–brain barrier breakdown after experimental stroke. Stroke 2013, 44:2536-2544.
• [39]Dimitrijevic OB, Stamatovic SM, Keep RF, Andjelkovic AV: Effects of the chemokine CCL2 on blood–brain barrier permeability during ischemia-reperfusion injury. J Cereb Blood Flow Metab 2006, 26:797-810.
• [40]Ge S, Song L, Serwanski DR, Kuziel WA, Pachter JS: Transcellular transport of CCL2 across brain microvascular endothelial cells. J Neurochem 2008, 104:1219-1232.
• [41]Etemad S, Zamin RM, Ruitenberg MJ, Filgueira L: A novel in vitro human microglia model: characterization of human monocyte-derived microglia. J Neurosci Methods 2012, 209:79-89.
• [42]Wu FY, Fan J, Tang L, Zhao YM, Zhou CC: Atypical chemokine receptor D6 inhibits human non-small cell lung cancer growth by sequestration of chemokines. Oncol Lett 2013, 6:91-95.
• [43]Fouillet A, Mawson J, Suliman O, Sharrack B, Romero IA, Woodroofe MN: CCL2 binding is CCR2 independent in primary adult human astrocytes. Brain Res 2012, 1437:115-126.