Study On The Alterations And Regulation Of Pro-Inflammatory Cytokines In SIRS Rat Models Generated By "Two-Hit

1. Establishment of the rat SIRS models by 'two-hit'.Objective To establish the stable Sprague-Dawley rat models of systemic inflammatory response syndrome (SIRS) by 'two-hit' that would imitate clinical features.Methods The rat models of SIRS were made by hemorrhagic shock as 'the first hit' and then with different doses of endotoxin as 'the second hit'. After 6 hours, changes of symptoms, oxygen pressure of arterial blood, white blood cell counts in peripheral blood, organ functions and the related pathology were analyzed.Results (1)After injuries, the body temperature of the experimental groups decreased,breath became faster, both oxygen pressure of arterial blood and the white blood cell counts in peripheral blood declined in different degrees (P<0.05 or <0.01). (2)The functional indexes of heart, liver and kidney were all increased markedly (P<0.05 or <0.01). (3)Results of pathological formation showed with multiple organ inflammatory changes.Conclusions With important values on studying the mechanism of SIRS, this model is an ideal animal model for studying SIRS at present.2. The changes of TNF- a and IL-6 at early stage in SIRS rat serum.Objective To inquire the changes of TNF- a and IL-6 in different time at early stage in SIRS rat serum and discuss the role of pro-inflammatory cytokines in the progress of SIRS.Methods The SIRS rat models were made following test one, and divided into the groups according to time phase. The rats were killed at 2h, 4h, 6h and 8h, the consistency of TNF- and IL-6 in serum were checked by enzyme-linked immunoadsordent assay(ELISA).Results The consistency of TNF- a in serum was increased obviously at once in2h,and reached to the peak in 4h,keeping high level in 6h and 8h;there were different changes between IL-6 and TNF- a , it displayed the tendency elevated step by step.Conclusions Keeping at high level at early stage of SIRS rat, TNF- a was risen to get up very important action in the SIRS occurrence and development, the IL-6 peaking lag, it confirmed IL-6 is not the main inflammatory cytokine causing the injury in early stage of SIRS.3. study on the activation of NF- B at early stage in rat SIRS modelsObjective To inquire the activation of NF- K B in Mononuclear cell(MNC) in blood in early stage of SIRS rat and discuss the roles of NF- K B in SIRS.Methods Mononuclear cells were separated from hemocytes from test two, and intracellular nucleoprotein were extracted, the activity of NF- K B were detected by electrophoretic mobility shift assay(EMSA).Results the activity of NF- K B was increased obviously at once in 2h than thecontrol group in MNC of blood in SIRS, keeping high lever in 4h,and began to decrease in 6h, and was near normal in 8h.Conclusions The activation of NF- K B was risen to get up very important action in regulating pro-inflammatory cytokines in early stage in SIRS rat.