Page - 112 - in Advances in Neuroimmunology

BrainSci. 2017,7, 14 morphine tolerancepredictedbyLINCS. In themorphine-tolerant+salineversusmorphine-tolerant+ LPSreport,AHR,UBE2L6,andPAFAH1B3were the topthreecandidates thatpotentially responsible for theLPS-inducedimmuneresponsesduringmorphine tolerance inrats. In theLINCSreports, the listedpotential targetshaddifferent rankingsusingthedifferentsetsofgenefeatures. Thisconfirms thattheresponsetoLPSbythoseinflammosome-relatedgenescouldbeaffectedbymorphinetolerance. Moreover,amongthe targets listedabove,whileVPS28wasNo. 1 in thecontrol rats inresponse toLPS, itwasNo. 4 inmorphine-tolerant rats (Table4). TheVPS28protein functions in transporting andsortingproteins intosub-cellularvesicles. Inourstudy,LINCSanalysis suggests that theactions ofVPS28 inresponse toLPScouldbedampenedduringmorphine tolerance. 5.Conclusions Theresults fromourstudyindicate that, in theratbrain,LPS-inducedinflammationinvolvesboth the inhibitionof theNLRP12anti-inflammatory inflammasomeandthestimulationofdownstream regulators such as Birc3, thereby increasing the expression of pro-inflammatory chemokines and cytokines.However, in themorphine tolerantstate, theresponse toLPSisdampened,as indicatedby thereducedexpressionof inflammasome-relatedgenes. LINCSanalysisconfirmedthat theresponseto LPSisalteredduringmorphinetoleranceandindicatedthatVPS28maybeoneofthegenesresponsible for thealterationsassociatedwithmorphine tolerance. Acknowledgments:Theauthors thankLouaineL.Spriggs forherexcellenteditorialassistance. Fundingfor this studywasprovided, inpart,byNational InstitutesofHealth (NIH)/National InstituteonDrugAbuse(NIDA) grantsR01DA007058andK02DA016149 toSulieL.Chang. Author Contributions: Sulie L. Chang designed the studies, participated in data collection, data analysis, andmanuscript preparation, andapproved themanuscript submission. WenfeiWangconducted theLINCS analysisandparticipated inmanuscriptpreparation. SabroniSarkarparticipated in thePCRarraydataanalysis andinmanuscriptpreparation.XinMaconductedtheanimal treatments, tissuecollection,andPCRarrayanalysis. Conflictsof Interest:Theauthorsdeclarenoconflictsof interest. AppendixA TableA1.GenesanalyzedinPCRArray. GeneSymbol FullName mRNAEntry Card6 caspaserecruitmentdomainfamily,member6 NM_001106413.1 Casp1 caspase1 NM_012762.2 Casp12 caspase12 NM_130422.1 Casp8 caspase8 NM_022277.1 Naip2 NLRfamily,apoptosis inhibitoryprotein6 XM_008760697.2 Nlrp12 NLRfamily,pyrindomaincontaining12 NM_001169142.1 Nlrp5 NLRfamily,pyrindomaincontaining5 NM_001107474.1 Nlrc4 NLRfamily,CARDdomaincontaining4 NM_001309432.1 Nlrp1a NLRfamily,pyrindomaincontaining1A NM_001145755.2 Nlrp3 NLRfamily,pyrindomaincontaining3 NM_001191642.1 Nlrp6 NLRfamily,pyrindomaincontaining6 NM_134375.3 Nlrx1 NLRfamilymemberX1 NM_001025010.1 Nod2 nucleotide-bindingoligomerizationdomaincontaining2 NM_001106172.1 Pycard PYDandCARDdomaincontaining NM_172322.1 Bcl2 BCL2,apoptosis regulator NM_016993.1 Bcl2l1 BCL2like1 NM_001033670.1 Birc2 baculoviral IAPrepeat-containing2 NM_021752.2 Birc3 baculoviral IAPrepeat-containing3 NM_023987.3 Cflar CASP8andFADD-likeapoptosis regulator NM_001033864.2 Chuk conservedhelix-loop-helixubiquitouskinase NM_001107588.1 Ciita class II,majorhistocompatibilitycomplex, transactivator NM_001270803.1 Ctsb cathepsinB NM_022597.2 Fadd Fasassociatedviadeathdomain NM_152937.2 Hsp90aa1 heatshockprotein90,alpha(cytosolic), classAmember1 NM_175761.2 Hsp90ab1 heatshockprotein90alphafamilyclassBmember1 NM_001004082.3 Ikbkb inhibitorofkappa lightpolypeptidegeneenhancer inB-cells,kinasebeta NM_053355.2 Ikbkg inhibitorofkappa lightpolypeptidegeneenhancer inB-cells,kinasegamma NM_199103.1 Irak1 interleukin-1receptor-associatedkinase1 NM_001127555.1 Map3k7 mitogenactivatedproteinkinasekinasekinase7 NM_001107920.2 Map3k7ip1 TGF-betaactivatedkinase1/MAP3K7bindingprotein1 NM_001109976.2 Map3k7ip2 TGF-betaactivatedkinase1/MAP3K7bindingprotein2 NM_001012062.1 Mapk1 mitogenactivatedproteinkinase1 NM_053842.2 Mapk11 mitogen-activatedproteinkinase11 NM_001109532.2 112