Study Of Signal Transduction Mechanisms Of Pbmc After Severely Burns

l. BackgroundBurn or trauma will not only induce local effect but also systemic responses inremote areas. Although many major improvements have been made in the treatment ofsevere burns, however, the prognosis is still far from ideal in sepsis or SIRS induceedby burn shock and systemic infection. Formally researchers were fond of blockingspecific cytokines in order to decrease the incidence SIRS in severe burns. However,the effect of treating a single factor was far from ideal since the interaction among thecytokines, e.g. the inflammatory mediators are complex and systematic.External stimulate will initiate stress reaction which might induce rapid systemicalterations. Such kind of reaction will elevate the concentration on intracellular signalmolecule which in turn transfer the signal into the cells, inducing the alterations ofgene expression leading to the alterations of specific protein levels. During the courseof signal transduction the number of nuclear transcription factor is less, however,transcription pathway mediated with diversity of biological effect.Nuclear factor-κB is one of the important transcription factor with very importanteffect in the rapid response system. The early gene mediated mainly partcipate thedefensive reaction of the boby. The main pathway of nuclear factor-κB mainly consistsproteokinases and low concentration OFRs mediators. There is close relationshipbetween the proteokinases and the changes of intracellular free Ca2+. After entering thecell, nuclear gene-κB combines the target gene sequence, where gene transcription isinitiated. The nuclear gene-κB dependent genes are proinflammatory factors andadhensive molecule, etc. There are direct relationships between proinflammatoryfactors and adhensive molecule with SIRS and MODS postburn.7The present study was designed to observe the changes of cellular signal molecules with determination of pro-inflammation cell gene expression. Application of calcium channel blocking agent and anti-oxidant regulates signal molecule and cytokine mRNA expression level. In order to elvaluate the effect of signal transcription pathway of nuclear gene kappa B in the pathogenesis of SIRS.2.Objective(1) To study the alteration of total plasma antioxidant capacity(TAC), NO and IL-6 in severely burned rats and to evaluate the relationship among NO, IL-6 and TAC. In order to demonstrate the expression and secretion of blood immune cell mediated inflammatory medium.(2) To study the influence of PBMC signal molecule by postburn plasma, in order to eveluate the regulating effect calcium channel blocking agent and antioxidant to signal molecule. In vitro study demonstrated that postburn plasma can act directly onto PBMCsignal molecule whose alterations can directly lead to cell preduced expression of inflammatory medium.(3)To study the regular alterations of expression of NF-KIB, FAK protein in PBMC, in order to clearify signal transcription of NF-icB concerned in the mechanism of inflammatory reaction in severe burns. Thereby, to observe the regulating effect of calcium channel blocking agent and antioxidant on nuclear factor-icB, and verify the medial effect of upstream signal molecules on NF-tcB activation.(4)To evaluate mainly the alterations of expression of NF-icB dependent proinflammatory cytokines mRNA and the alterations of systemic inflammation reaction and immuno-functjon closely related ThlITh2 cytokines. Thereby, to verify the regulating effect of expression of cytokines by calcium channels blocker and antioxidant.83.Materials and methodsSprague Dawley rats were randomly assigned to each group. A 30% TBSA full-thickness scald was performed by immersing in 1000C water for 12 seconds. Plasma and PBMC were isolated at different time point after scalding. Sham-burned controls were treated in the same way instead of immersing in 370C water.In vitro study, PBMC of the normal rats were isolated and culture with post-bum serum (2, 6, 8, 12, 24, 48h BS ).