Page - 106 - in Advances in Neuroimmunology

BrainSci. 2017,7, 14 Table 1. Expression profile of inflammasomes and NLR genes in the rat brain in response to lipopolysaccharides (LPS),withandwithoutmorphine tolerance. morphine-tolerant+ saline/placebocontrol+saline placebocontrol+LPS/placebo control+saline morphine-tolerant+ LPS/morphine-tolerant+saline Gene FoldChange p-value* FoldChange** p-value* FoldChange** p-value* Card6 −1.2347 0.078926 −1.5292 0.004159 −1.714 0.017499 Casp1 1.1505 0.299299 1.2132 0.276593 1.2764 0.02969 Casp12 1.0972 0.462347 1.5422 0.003614 1.4249 0.092085 Casp8 −1.0402 0.612597 −1.1363 0.391056 −1.2652 0.139999 Naip2 −1.4483 0.067329 −1.1031 0.619799 −1.2316 0.25758 Nlrp12 −1.6504 0.674422 −7.31 0.002731 −4.5124 0.025372 Nlrp5 1.494 0.054876 1.274 0.20786 1.2819 0.027352 Nlrc4 1.0028 0.915792 1.1349 0.159712 1.1003 0.445719 Nlrp1a −1.0898 0.470844 −1.2027 0.001147 −1.3308 0.1379 Nlrp3 1.3773 0.134647 1.5114 0.054565 1.3956 0.02407 Nlrp6 1.205 0.420019 −1.6225 0.358997 1.0293 0.784547 Nlrx1 −1.1574 0.24832 −1.1494 0.191026 1.039 0.679959 Nod2 −1.6257 0.650152 1.0901 0.811946 1.2826 0.50395 Pycard −1.0575 0.610143 −1.0285 0.797363 −1.2363 0.030115 *Forp-value, letters inredmeanp<0.05. **ForFoldChange, letters inredmeanfoldchange>2andletters inblue mean the foldchange<−2. For the color lines, : 6–10 folddecrease; : 2–5 folddecrease; : <2 fold; : 2–5 fold increase; : 6–10 fold increase; : 11–30 fold increase; : 31–50 fold increase. 3.2. ExpressionProfile of Inflammasome-RelatedDownstreamSignalingGenesFollowinganLPSChallenge, WithandWithoutMorphineTolerance With a few exceptions, there were no significant changes in expression of the downstream signalinggenes inresponse toLPSineither themorphine-tolerant rats (morphine-tolerant+LPS)or thecontrolanimals (placebo-control+LPS), comparedto theratsgivensaline (morphine-tolerant+ salineandplacebo-control+saline) (Table1).However,Baculoviral IAPRepeat-Containing3(Birc3), an important regulatorgene involvedin thedownstreameffectsof inflammasomes,wassignificantly increasedinresponse toLPSinboththemorphine-tolerant (21.5 fold;p<0.05)andcontrol (21 fold; p< 0.01) groups, compared to the rats given saline (Figure 2, Table 2). In addition,NF-Kappa-B InhibitorAlpha (Nfkbia), an inhibitor protein ofNF-κB,was increased in both groups givenLPS (morphine-tolerant+LPS,3.4 fold,p<0.01;placebo-control+LPS,3.9 fold,p<0.01) (Figure2,Table2). Pla ce bo -co ntr ol+ Sa lin e Pla ce bo -co ntr ol+ LP S Mo rp hin e-t ole ran t+S ali ne Mo rp hin e-t ole ran t+L PS -5 0 5 10 15 20 25 Birc3 Nfkbi Figure 2. Inflammasome-related downstream gene expression in the rat brain in response to Lipopolysaccharides (LPS), with and without morphine tolerance. The expression of the inflammasome-relateddownstreamsignalinggenesBaculoviral IAPRepeat-Containing3(Birc3)and NF-Kappa-BInhibitorAlpha(Nfkbia) in thebrainsof ratsgivenani.p. injectionofeither250μg/kg LPSorsaline,withandwithoutmorphine tolerance (n=3–5ratspergroup),wasdeterminedusinga PolymeraseChainReaction(PCR)array. ThedatawerecalculatedusingtheΔΔCTmethodrelative to thecontrolgroup(placebo-control+saline)andarerepresentedas foldchange. 106