| ObjectiveTo study the expression of the effecter molecule of myocardial cell apoptosis (caspase-3),the resisting apoptosis protein(bcl-2) and the relevance of inflammatory mediators TNF-αand myocardial apoptosis index AI,revealed the role of TNF-αin myocadiocyte apoptosis.giving the intervention effects of anti-TNFα-mAb to rectify the causal relation of TNF-αand AI.Materials and Methods72 wistar rats were randomly divided into control group(n=24),septic group(n=24)and interference group(n=24),either group was then divided into four subgroups:3h,12h,24h,36h,the sepsis model was made by cecal ligation and puncture(CLP)in septic group,control group got the same treatments except CLP,the interference group was injected anti-TNF-α-mab(0.2mg/kg) into abdominal cavity 30 minute before CLP.ELISA were used to determined TNF-α,Tunel were used to determined AI,western blot were used to determined caspase-3 and bcl-2 expression.Results1.Pathological changesLight microscope:in the control group there was no pathological changes,the cardiac muscle fibers lined up regularly,the structure was clear.septic group with the time,the pathological changes were gradully appeared,the cardiac muscle fibers were chaotic,blood ressels were hyperema and hemorrhage,we can see piles of blood cells among the myocardial cells,especially in 36h,Myocardial cell swelling or shrinkage, nuclear deformation.In interference group the cardiac muscle fibers were chaotic,we can see petechia,pathological changes were Weakened compared to septic group At the same time point,no significant changes in the nucleus.2.enzymelinked immunosorbentassy(ELISA)TNF-αprotein levels was significantly increased compared to normal control in myocardium tissure(P＜0.01),36h group was the highest,but there was no significant difference between 24h and 36h(P＞0.05),TNF-αprotein levels was significantly decreased compared to septic group in interference group(P＜0.01).3.TdT-mediated dUTP nick end labeling(TUNEL)the number of myocardial apoptosis was higher in CLP than in control group(P＜0.01)and progressively increased.the index of myocardial apoptosis was in 36h group and there was no significant difference between 24h and 36h(P＞0.05),the number was decreased compared to septic group in interference group(P＜0.01).4.Westen blotthe same hour of caspase-3 was higher in septic and interference group than in contol group(P＜0.01)and the level of caspase-3 was higher in septic group than in interference group(P＜0.01).the same hour of bcl-2 was lower in septic and interference group than in contol group(P＜0.01)and the level of bcl-2 was higher in interference group than in septic group(P＜0.01)5.correlation analysisthere was positive correlation between TNF-αand AI(r=0.903,P＜0.001).ConclusionIn the present study,the model of septic rats was made by cecal ligation and puncture(CLP),after the Surgery was given for 3h,the rats showed up the changes of general state,WBC and so on,these Performance was consistent with sepsis,the model can be certified be Successful,the observation of caspase-3,bcl-2and TNF-αshowed that The development of sepsis,Myocardial Apoptosis gradually increased with the development of sepsis,there was positive correlation between TNF-αand AI,TNF-a play an important role in myocardial apoptosis,The use of anti-TNF-α-mab pre-treatment of rats made this results directly.these data suggest that TNF-αmay be one of mechanisms about myocardial apoptosis in sepsis.TNF-α,bcl-2 and caspase-3 protein may be used to be a sign in the change of pathogenetic condition,we can use them to iprove the prognosis in sepsis。...
|
PDF Full Text Request