Page - 109 - in Advances in Neuroimmunology

BrainSci. 2017,7, 14 LPS),whereas in themorphine-tolerant (morphine-tolerant+LPS)groupthe foldchanges (9-, 7-, 14-, and3-fold, respectively)werenotstatisticallysignificant (Figure4,Table3). Figure 4. Chemokine gene expression in the rat brain in response to Lipopolysaccharides (LPS), withandwithoutmorphine tolerance.Geneexpressionof thechemokinesC-Cmotif chemokine ligand (Ccl)2,Ccl5,Ccl7,Ccl11,Ccl12,C-X-Cmotif chemokine ligand(Cxcl)1,andCxcl3 in thebrainsof rats withandwithoutmorphine tolerance, followingan i.p. injectionof either250μg/kgLPSor saline (n=3–5ratspergroup),wasdeterminedusingaPolymeraseChainReaction(PCR)array.Datawere calculatedusing theΔΔCTmethod relative to the control group (placebo-control+ saline) andare representedasa foldchange. *p<0.05, **p<0.01, ***p<0.001 3.4. LINCSAnalysis of theDifferentiallyExpressedGenes Differentiallyexpressedgenes in themorphine-tolerant+salineversusmorphine-tolerant+LPS ratsandintheplacebo-control+salineversusplacebo-control+LPSratsaswellasgenechanges in rats theplacebo-control+ salineversusmorphine-tolerant+ saline ratswere input into theQuery App(apps.lincscloud.org/query). OnereportwasgeneratedbyLINCSforeachsetofgenes input. Thegeneswithahighpositivescore inConsensusKnockdownConnectionswereconsideredtobe potentialgenetargets (Table4). In theplacebo-control+salineversusplacebo-control+LPSreport, VPS28,proteinCreceptor (PROCR),andchargedmultivesicularbodyprotein2A(CHMP2A)were the topthreewith thehighest scores.VPS28 isanESCRT-Icomplexsubunit that functions in the transport andsortingofproteins intosub-cellularvesicles. PROCRisendothelialproteinCreceptor involved in thebloodcoagulationpathway.CHMP2Aisacomponentof theendosomalsortingcomplexrequired for transport III,which is involvedinthedegradationofsurfacereceptorproteinsandtheformationof endocyticmultivesicularbodies. In the placebo-control + saline versusmorphine-tolerant + saline report, SWI/SNF related, matrix associated, actin dependent regulator of chromatin, subfamily e, member 1 (SMARCE1), aryl-hydrocarbonreceptor repressor (AHRR), andglutathioneperoxidase7 (GPX7)were themost likely targetspredictedbyLINCS.SMARCE1is requiredfor the transcriptionalactivationofgenes normallyrepressedbychromatin.AHRRmediatesdioxin toxicityandis involved in theregulationof cellgrowthanddifferentiation.GPX7is involvedwithcellularsenescenceandinsulinsecretion. In themorphine-tolerant+salineversusmorphine-tolerant+LPSgroup,AHR(arylhydrocarbon receptor),UBE2L6(ubiquitin-conjugatingenzymeE2L6),andPAFAH1B3(platelet-activatingfactor acetylhydrolase 1b, Catalytic Subunit 3) were the top three candidates. AHR is involved in the regulation of biological responses to planar aromatic hydrocarbons; UBE2L6 targets abnormal or short-livedproteins fordegradation;andPAFAH1B3functions inbraindevelopmentandisassociated withmental retardation,ataxia,andatrophyof thebrain. 109