Septic shock is the most common cause of death in intensive care units due to an aggressive inflammatory response that leads to multiple organ failure. However, a lipopolysaccharide (LPS) tolerance phenomenon (a nonreaction to LPS), is also often described. Neither the inflammatory response nor the tolerance is completely understood. In this work, both of these responses were analyzed using microarrays in zebrafish. Fish that were 4 or 6 days postfertilization (dpf) and received a lethal dose (LD) of LPS exhibited 100% mortality in a few days. Their transcriptome profile, even at 4 dpf, resembled the profile in humans with severe sepsis. Moreover, we selected 4-dpf fish to set up a tolerance protocol: fish treated with a nonlethal concentration of Escherichia coli LPS exhibited complete protection against the LD of LPS. Most of the main inflammatory molecules described in mammals were represented in the zebrafish microarray experiments. Additionally and focusing on this tolerance response, the use of cyclodextrins may mobilize cholesterol reservoirs to decrease mortality after a LD dose of LPS. Therefore, it is possible that the use of the whole animal could provide some clues to enhance the understanding of the inflammatory/tolerance response and to guide drug discovery.
Get full access to this article
View all access options for this article.
References
1.
GuhaM, MackmanN. LPS induction of gene expression in human monocytes. Cell Signal, 2001; 13:85–94.
2.
AngusDC, WaxRS. Epidemiology of sepsis: an update. Crit Care Med, 2001; 29(Suppl 7):S109–S116.
MarshallJC. Inflammation, coagulopathy, and the pathogenesis of multiple organ dysfunction syndrome. Crit Care Med, 2001; 29:S99–S106.
5.
DavisJM, ClayH, LewisJL, GhoriN, HerbomelP, RamakrishnanL. Real-time visualization of mycobacterium-macrophage interactions leading to initiation of granuloma formation in zebrafish embryos. Immunity, 2002; 17:693–702.
6.
LamSH, ChuaHL, GongZ, LamTJ, SinYM. Development and maturation of the immune system in zebrafish, Danio rerio: a gene expression profiling, in situ hybridization and immunological study. Dev Comp Immunol, 2004; 28:9–28.
7.
ReddMJ, KellyG, DunnG, WayM, MartinP. Imaging macrophage chemotaxis in vivo: studies of microtubule function in zebrafish wound inflammation. Cell Motil Cytoskeleton, 2006; 63:415–422.
8.
LieschkeGJ, CurriePD. Animal models of human disease: zebrafish swim into view. Nat Rev Genet, 2007; 8:353–367.
9.
TredeNS, OtaT, KawasakiH, PawBH, KatzT, DemarestB, et al.Zebrafish mutants with disrupted early T-cell and thymus development identified in early pressure screen. Dev Dyn, 2008; 237:2575–2584.
10.
StockhammerOW, ZakrzewskaA, HegedûsZ, SpainkHP, MeijerAH. Transcriptome profiling and functional analyses of the zebrafish embryonic innate immune response to Salmonella infection. J Immunol, 2009; 182:5641–5653.
11.
WesterfieldM. The Zebrafish Book. A Guide for the Laboratory Use of Zebrafish (Danio rerio). Eugene: University of Oregon Press, 2000.
12.
Nusslein-VolhardC, DahmR. Zebrafish, a Practical Approach. Oxford: Oxford University Press, 2002.
13.
NovoaB, BowmanTV, ZonL, FiguerasA. LPS response and tolerance in the zebrafish (Danio rerio). Fish Shellfish Immunol, 2009; 26:326–331.
BrazmaA, HingampP, QuackenbushJ, SherlockG, SpellmanP, StoeckertC, et al.Minimum information about a microarray experiment (MIAME)-toward standards for microarray data. Nat Genet, 2001; 29:365–371.
ConesaA, GotzS, Garcia-GomezJM, TerolJ, TalonM, RoblesM. Blast2GO: a universal tool for annotation, visualization and analysis in functional genomics research. Bioinformatics, 2005; 21:3674–3676.
18.
AlexeyenkoA, WassenbergDM, LobenhoferEK, YenJ, LinneyE, SonnhammerEL, et al.Dynamic zebrafish interactome reveals transcriptional mechanisms of dioxin toxicity. PLoS One, 2010; 5:e10465.
19.
MontojoJ, ZuberiK, RodriguezH, KaziF, WrightG, DonaldsonSL, et al.GeneMANIA Cytoscape plugin: fast gene function predictions on the desktop. Bioinformatics (Oxford, England), 2010; 26:2927–2928.
20.
SmootME, OnoK, RuscheinskiJ, WangPL, IdekerT. Cytoscape 2.8: new features for data integration and network visualization. Bioinformatics, 2011; 27:431–432.
21.
MaereS, HeymansK, KuiperM. BiNGO: a Cytoscape plugin to assess overrepresentation of gene ontology categories in biological networks. Bioinformatics (Oxford, England), 2005; 21:3448–3449.
22.
MorrisJH, ApeltsinL, NewmanAM, BaumbachJ, WittkopT, SuG, et al.ClusterMaker: a multi-algorithm clustering plugin for Cytoscape. BMC Bioinformatics, 2011; 12:436.
23.
BarskyA, GardyJL, HancockRE, MunznerT. Cerebral: a Cytoscape plugin for layout of and interaction with biological networks using subcellular localization annotation. Bioinformatics, 2007; 23:1040–1042.
24.
RozenS, SkaletskyH. Primer3 on the www for general users and for biologist programmers. Methods Mol Biol, 2000; 132:365–386.
25.
PfafflMW. A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res, 2001; 29:e45.
26.
WillettCE, CortesA, ZuastiA, ZapataAG. Early hematopoiesis and developing lymphoid organs in the zebrafish. Dev Dyn, 1999; 214:323–336.
HasselgrenPO, MenconiMJ, FareedMU, YangH, WeiW, EvensonA. Novel aspects on the regulation of muscle wasting in sepsis. Int J Biochem Cell Biol, 2005; 37:2156–2168.
29.
HungCC, LiuX, KwonMY, KangYH, ChungSW, PerrellaMA. Regulation of heme oxygenase-1 gene by peptidoglycan involves the interaction of Elk-1 and C/EBPalpha to increase expression. Am J Physiol Lung Cell Mol Physiol, 2010; 298:L870–L879.
30.
HuaF, WangJ, IshratT, WeiW, AtifF, SayeedI, et al.Genomic profile of Toll-like receptor pathways in traumatically brain-injured mice: effect of exogenous progesterone. J Neuroinflamm, 2011; 8:42.
CynisH, HoffmannT, FriedrichD, KehlenA, GansK, KleinschmidtM, et al.The isoenzyme of glutaminyl cyclase is an important regulator of monocyte infiltration under inflammatory conditions. EMBO Mol Med, 2011; 3:545–558.
ZhangG, WangJ, KellyJ, GuG, HouJ, ZhouY, et al.B7-H3 Augments the inflammatory response and is associated with human sepsis. J Immunol, 2010; 185:3677–3684.
35.
FassbenderM, GerlitzkiB, UllrichN, LuppC, KleinM, RadsakMP, et al.Cyclic adenosine monophosphate and IL-10 coordinately contribute to nTreg cell-mediated suppression of dendritic cell activation. Cell Immunol, 2010; 265:91–96.
36.
LiS, MiaoT, SebastianM, BhullarP, GhaffariE, LiuM, et al.The transcription factors Egr2 and Egr3 are essential for the control of inflammation and antigen-induced proliferation of B and T cells. Immunity, 2012; 37:685–696.
37.
NguyenHB, LoombaM, YangJJ, JacobsenG, ShahK, OteroRM, et al.Early lactate clearance is associated with biomarkers of inflammation, coagulation, apoptosis, organ dysfunction and mortality in severe sepsis and septic shock. J Inflamm (Lond), 2010; 7:6.
38.
DingY, KantarciA, BadweyJA, HasturkH, MalabananA, Van DykeTE. Phosphorylation of pleckstrin increases proinflammatory cytokine secretion by mononuclear phagocytes in diabetes mellitus. J Immunol, 2007; 179:647–654.
39.
GrewalPK, UchiyamaS, DittoD, VarkiN, LeDT, NizetV, et al.The Ashwell receptor mitigates the lethal coagulopathy of sepsis. Nat Med, 2008; 14:648–655.
MakowskiL, HotamisligilGS. Fatty acid binding proteins-the evolutionary crossroads of inflammatory and metabolic responses. J Nutr, 2004; 134:2464S–2468S.
42.
AzizM, JacobA, YangWL, MatsudaA, WangP. Current trends in inflammatory and immunomodulatory mediators in sepsis. J Leukoc Biol, 2013; 93:329–342.
43.
DasUN. Is sepsis a pro-resolution deficiency disorder?. Med Hypotheses, 2013; 80:297–299.
44.
LiL, HossainMA, SadatS, HagerL, LiuL, TamL, et al.Lecithin cholesterol acyltransferase null mice are protected from diet-induced obesity and insulin resistance in a gender-specific manner through multiple pathways. J Biol Chem, 2011; 286:17809–17820.
45.
BaumanDR, BitmansourAD, McDonaldJG, ThompsonBM, LiangG, RussellDW. 25-Hydroxycholesterol secreted by macrophages in response to Toll-like receptor activation suppresses immunoglobulin A production. Proc Natl Acad Sci U S A, 2009; 106:16764–16769.
46.
DiczfalusyU, OlofssonKE, CarlssonAM, GongM, GolenbockDT, RooyackersO, et al.Marked upregulation of cholesterol 25-hydroxylase expression by lipopolysaccharide. J Lipid Res, 2009; 50:2258–2264.
47.
ParkK, ScottAL. Cholesterol 25-hydroxylase production by dendritic cells and macrophages is regulated by type I interferons. J Leukoc Biol, 88; 1081–1087.
StrassheimD, ParkJS, AbrahamE. Sepsis: current concepts in intracellular signaling. Int J Biochem Cell Biol, 2002; 34:1527–1533.
50.
HaskóG, LindenJ, CronsteinB, PacherP. Adenosine receptors: therapeutic aspects for inflammatory and immune diseases. Nat Rev Drug Discov, 2008; 7:759–770.
51.
CsókaB, NémethZH, RosenbergerP, EltzschigHK, SpolaricsZ, PacherP, et al.A2B adenosine receptors protect against sepsis-induced mortality by dampening excessive inflammation. J Immunol, 2010; 185:542–550.
52.
KronlageM, SongJ, SorokinL, IsfortK, SchwerdtleT, LeipzigerJ, et al.Autocrine purinergic receptor signaling is essential for macrophage chemotaxis. Sci Signal, 2010; 3:ra55.
53.
AherneCM, KewleyEM, EltzschigHK. The resurgence of A2B adenosine receptor signaling. Biochim Biophys Acta Biomembr, 2011; 1808:1329–1339.
54.
FerreroME. Purinoceptors in inflammation: potential as anti-inflammatory therapeutic targets. Front Biosci, 2011; 17:2172–2186.
55.
RamakersBP, RiksenNP, van der HoevenJG, SmitsP, PickkersP. Modulation of innate immunity by adenosine receptor stimulation. Shock, 2011; 36:208–215.
56.
DongC, ZhangX, ZhouF, DouH, DuvernayMT, ZhangP, et al.ADP-ribosylation factors modulate the cell surface transport of G protein-coupled receptors. J Pharmacol Exp Ther, 2010; 333:174–183.
57.
NovoaB, FiguerasA. Zebrafish: model for the study of inflammation and the innate immune response to infectious diseases. Adv Exp Med Biol, 2012; 946:253–275.
58.
van der Pouw KraanTC, SnijdersA, BoeijeLC, de GrootER, AlewijnseAE, LeursR, et al.Histamine inhibits the production of interleukin-12 through interaction with H2 receptors. J Clin Invest, 1998; 102:1866–1873.
59.
CaronG, DelnesteY, RoelandtsE, DuezC, HerbaultN, MagistrelliG, et al.Histamine induces CD86 expression and chemokine production by human immature dendritic cells. J Immunol, 2001; 166:6000–6006.
60.
MazzoniA, YoungHA, SpitzerJH, VisintinA, SegalDM. Histamine regulates cytokine production in maturing dendritic cells, resulting in altered T cell polarization. J Clin Invest, 2001; 108:1865–1873.
61.
MorichikaT, TakahashiHK, IwagakiH, YoshinoT, TamuraR, YokoyamaM, et al.Histamine inhibits lipopolysaccharide-induced tumor necrosis factor-alpha production in an intercellular adhesion molecule-1- and B7.1-dependent manner. J Pharmacol Exp Ther, 2003; 304:624–633.
62.
TakahashiHK, IwagakiH, MoriS, YoshinoT, TanakaN, NishiboriM. Histamine inhibits lipopolysaccharide-induced interleukin (IL)-18 production in human monocytes. Clin Immunol, 2004; 112:30–34.
63.
JutelM, AkdisM, AkdisCA. Histamine, histamine receptors and their role in immune pathology. Clin Exp Allergy, 2009; 39:1786–1800.
64.
PengJ, YuanQ, LinB, PanneerselvamP, WangX, LuanXL, et al.SARM inhibits both TRIF- and MyD88-mediated AP-1 activation. Eur J Immunol, 2010; 40:1738–1747.
65.
JiangG, ZhangBB. Glucagon and regulation of glucose metabolism. Am J Physiol Endocrinol Metab, 2003; 284:E671–E678.
66.
JeschkeMG, KleinD, BolderU, EinspanierR. Insulin attenuates the systemic inflammatory response in endotoxemic rats. Endocrinology, 2004; 145:4084–4093.
67.
RussellJA. Management of sepsis. N Engl J Med, 2006; 355:1699–1713.
68.
ShapiroNI, HowellMD, TalmorD, LaheyD, NgoL, BurasJ, et al.Implementation and outcomes of the multiple urgent sepsis therapies (MUST) protocol. Crit Care Med, 2006; 34:1025–1032.
69.
SatoY, NishioY, SekineO, KodamaK, NagaiY, NakamuraT, et al.Increased expression of CCAAT/enhancer binding protein-beta and -delta and monocyte chemoattractant protein-1 genes in aortas from hyperinsulinaemic rats. Diabetologia, 2007; 50:481–489.
70.
AuthierF, DesbuquoisB. Glucagon receptors. Cell Mol Life Sci, 2008; 65:1880–1899.
SchusterH. High risk/high priority: familial hypercholesterolemia—a paradigm for molecular medicine. Atheroscler Suppl, 2002; 2:27–30; discussion 30–32.
75.
TominagaK, SaitoS, MatsuuraM, NakanoM. Lipopolysaccharide tolerance in murine peritoneal macrophages induces downregulation of the lipopolysaccharide signal transduction pathway through mitogen-activated protein kinase and nuclear factor-κ B cascades, but not lipopolysaccharide-incorporation steps. Biochim Biophys Acta, 1999; 1450:130–144.
76.
MedvedevAE, KopydlowskiKM, VogelSN. Inhibition of lipopolysaccharide-induced signal transduction in endotoxin-tolerized mouse macrophages: dysregulation of cytokine, chemokine, and Toll-like receptor 2 and 4 gene expression. J Immunol, 2000; 164:5564–5574.
77.
MedvedevAE, LentschatA, WahlLM, GolenbockDT, VogelSN. Dysregulation of LPS-induced Toll-like receptor 4-MyD88 complex formation and IL-1 receptor-associated kinase 1 activation in endotoxin-tolerant cells. J Immunol, 2002; 169:5209–5216.
78.
DobrovolskaiaMA, VogelSN. Toll receptors, CD14, and macrophage activation and deactivation by LPS. Microbes Infect, 2002; 4:903–914.
79.
DobrovolskaiaMA, MedvedevAE, ThomasKE, CuestaN, ToshchakovV, RenT, et al.Induction of in vitro reprogramming by Toll-like receptor (TLR)2 and TLR4 agonists in murine macrophages: effects of TLR “homotolerance” versus “heterotolerance” on NF-κ B signaling pathway components. J Immunol, 2003; 170:508–519.
80.
RenshawSA, LoynesCA, TrushellDM, ElworthyS, InghamPW, WhyteMK. A transgenic zebrafish model of neutrophilic inflammation. Blood, 2006; 108:3976–3978.
81.
BaatarD, PatelK, TaubDD. The effects of ghrelin on inflammation and the immune system. Mol Cell Endocrinol, 2011; 340:44–58.
82.
FoellD, WittkowskiH, RenZ, TurtonJ, PangG, DaebritzJ, et al.Phagocyte-specific S100 proteins are released from affected mucosa and promote immune responses during inflammatory bowel disease. J Pathol, 2008; 216:83–192.
83.
ManolakisAC, KapsoritakisAN, GeorgouliasP, TzavaraC, ValotassiouV, KapsoritakiA, et al.Moderate performance of serum S100A12, in distinguishing inflammatory bowel disease from irritable bowel syndrome. BMC Gastroenterol, 2010; 10:118.
ShenSS, CallaghanD, JuzwikC, XiongH, HuangP, ZhangW. ABCG2 reduces ROS-mediated toxicity and inflammation: a potential role in Alzheimer's disease. J Neurochem, 2010; 114:1590–1604.
86.
OlsonN, HristovaM, HeintzNH, LounsburyKM, Van Der VlietA. Activation of hypoxia-inducible factor-1 protects airway epithelium against oxidant-induced barrier dysfunction. Am J Physiol Lung Cell Mol Physiol, 2011; 301:L993–L1002.
87.
KaoKK, FinkMP. The biochemical basis for the anti-inflammatory and cytoprotective actions of ethyl pyruvate and related compounds. Biochem Pharmacol, 2010; 80:151–159.
88.
VillafuerteBC, KaytorEN. An insulin-response element-binding protein that ameliorates hyperglycemia in diabetes. J Biol Chem, 2005; 280:20010–20020.
89.
VillafuerteBC, BaratiMT, SongY, MooreJP, EpsteinPN, PortilloJ. Transgenic expression of insulin-response element binding protein-1 in beta-cells reproduces type 2 diabetes. Endocrinology, 2009; 150:2611–2617.
PeakePW, ShenY, CampbellLV, CharlesworthJA. Human adiponectin binds to bacterial lipopolysaccharide. Biochem Biophys Res Commun, 2006; 341:108–115.
92.
BenoitME, TennerAJ. Complement protein C1q-mediated neuroprotection is correlated with regulation of neuronal gene and microRNA expression. J Neurosci, 2011; 31:3459–3469.
93.
ChinnaiyanAM, Huber-LangM, Kumar-SinhaC, BarretteTR, Shankar-SinhaS, SarmaVJ, et al.Molecular signatures of sepsis: multiorgan gene expression profiles of systemic inflammation. Am J Pathol, 2001; 159:1199–1209.
94.
CalvanoSE, XiaoW, RichardsDR, FelcianoRM, BakerHV, ChoRJ, et al.A network-based analysis of systemic inflammation in humans. Nature, 2005; 437:1032–1037.
95.
PachotA, LepapeA, VeyS, BienvenuJ, MouginB, MonneretG. Systemic transcriptional analysis in survivor and non-survivor septic shock patients: a preliminary study. Immunol Lett, 2006; 106:63–71.
96.
JohnsonSB, LissauerM, BochicchioGV, MooreR, CrossAS, ScaleaTM. Gene expression profiles differentiate between sterile SIRS and early sepsis. Ann Surg, 2007; 245:611–621.
97.
ZhuH, TangY, IvanciuL, CentolaM, LupuC, TaylorFBJr., et al.Temporal dynamics of gene expression in the lung in a baboon model of E. coli sepsis. BMC Genomics, 2007; 8:58.
98.
BhattyM, FanR, MuirWM, PruettSB, NanduriB. Transcriptomic analysis of peritoneal cells in a mouse model of sepsis: confirmatory and novel results in early and late sepsis. BMC Genomics, 2012; 13:509.
99.
LambeckS, WeberM, GonnertFA, MrowkaR, BauerM. Comparison of sepsis-induced transcriptomic changes in a murine model to clinical blood samples identifies common response patterns. Front Microbiol, 2012; 3:284.
100.
MasloveDM, TangBM, McLeanAS. Identification of sepsis subtypes in critically ill adults using gene expression profiling. Crit Care, 2012; 16:R183.
101.
StarrME, HuY, StrombergAJ, CarmicalJR, WoodTG, EversBM, et al.Gene expression profile of mouse white adipose tissue during inflammatory stress: age-dependent upregulation of major procoagulant factors. Aging Cell, 2013; 12:194–206.
102.
SimonsK, IkonenE. How cells handle cholesterol. Science, 2000; 290:1721–1726.
103.
SimonsK, ToomreD. Lipid rafts and signal transduction. Nat Rev Mol Cell Biol, 2000; 1:31–39.
Laza-KnoerrAL, GrefR, CouvreurP. Cyclodextrins for drug delivery. J Drug Target, 2010; 18:645–656.
106.
MuraP, CortiG, CirriM, MaestrelliF, MenniniN, BragagniM. Development of mucoadhesive films for buccal administration of flufenamic acid: effect of cyclodextrin complexation. J Pharm Sci, 2010; 99:3019–3029.
107.
DanciuC, SoicaC, CsanyiE, AmbrusR, FefleaS, PeevC, et al.Changes in the anti-inflammatory activity of soy isoflavonoid genistein versus genistein incorporated in two types of cyclodextrin derivatives. Chem Cent J, 2012; 6:58.
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
