| Objectives:Sepsis is an uncontrolled systemic inflammatory response syndrome(SIRS) caused by infection,which is a common complication in serious infection,injury,burning,major operations,severe pancreatitis and shock.It can lead to septic shock,acute respiratory distress syndrome(ARDS) and multiple organ disfunction syndrome(MODS).The morbility is rising by 1.5 to 8 percent in recent 30 years.The mortality reaches up to 40 percent.so it is the main clinical cause of death in critical illness.There is no breakthrough in sepsis treatment now,the reason is that the pathogenesis is complex,the content involves in a series of problems such as inflammation, immunity, blood coagulation and histologic lesion and so on,and sepsis has close relationship with pathology and physiology of multisystem and organ.The endothelial cells occupy the core position.It's the regulator of inflammatory cascade reaction and important composition of parasitifer to sepsis,the changes of structure and function are perhaps the central link in sepsis development,and then hasten circulation of inflammatory reaction, activation of coagulation system and intercellular interaction,finally lead to thrombogenesis,hyoxemia and organ dysfunction.So the activation and injury of endothelial cells have close relationship with sepsis.Abroad researches have certified that statins have the effect of antiinflammatory, anticoagulation,lessening injury of important organs and lower the morbility and mortality of sepsis for the past few years.Statins is becoming as the hot spot in researches and treatment of sepsis in recent years.One of the main mechanism may have relationship with protecting vessel endothelium function.In our research,the rat septic model was made by cecal ligation and puncture(CLP),and observe endothelium protection of Simvastatin in sepsis by setting up multiple time points,expecting to decrease complications and reduce mortality.So it can provide a theoretical basis for sepsis in clinical medication.Methods: Eighty clean female Wistar rats weight 200-250g (supplied by the experimental animal department of Hebei medical university) were randomly divided into four groups.GroupI:healthy control group(8 rats),groupII:sham operation group(8 rats),groupIII:model group(32 rats), groupIV:treatment group with Simvastatin(32 rats).The rat septic model was made by cecal ligation and puncture(CLP).The rats had the performances of tired spirit, abdominal distention, restlessness, shivers, increased canthal secretion, hairs in disorder after CLP.After executed,we saw the haematodes seepage in abdominal cavity,the swelling darkening and adhesive caecum,the inflatable jejunum and so on.All these certified that the rat septic models were made successfully.GroupIV was administered with Simvastatin (20mg/kg/day) before CLP by gastric tube for 2 weeks continuously,once daily.The other three groups were given saline of the same volume also via gastric tube for 2 weeks.The blood and aorta thoracalis specimens of groupI were taken after being killed;They were taken after sham operation for 6h in groupII and after CLP for 3h,6h,24h,48h in groupIII and groupIV respectively(8 rats in each subgroup).Taking rectal temperature and blood routine to help establish the models were successful;We observed the death status and calculated mortality in each group;Serum was taken to measure E-selectin and von Willebrand disease (vWF) by means of ELISA;The aorta thoracalis specimens were used to measure inducible nitric oxide synthase(iNOS) protein and endothelial nitric oxide synthase(eNOS) protein by Western Blotting.Results1 The general performance of rats: The rats in groupIII and groupIV may drink water freely in 2-3h after CLP,and had the performances of tired spirit,abdominal distention,restlessness,shivers,loose stools,increased canthal secretion,hairs in disorder,reduction of eating and drinking in 6-8h after CLP.After executed,we saw the haematodes seepage in abdominal cavity,the swelling darkening and adhesive caecum,the inflatable jejunum,hepato- pulmonary purulent,and something foul smell.The rats in groupII were normal in appearance and activity,they had no pathological changes in abdominal viscera and were all survival.2 WBC: The WBC in groupI was (6.78±0.94)×109/L,groupII was slightly higher than groupI(P>0.05);The WBC in groupIII was (10.43±0.66)×109/L at 3h after CLP,it rose up to (12.79±0.51)×109/L at 6h and showed downward trend since then,decreased to (8.52±0.40)×109/L at 24h,but all higher than groupI and groupII(P<0.05),the WBC decreased to (7.45±0.59)×109/L at 48h and lower significantly than 3h and 6h(P<0.05);The WBC in groupIV was (8.79±0.70)×109/L at 3h,(10.36±0.19)×109/L at 6h,they were all lower than groupIII at same point(P<0.05).3 Rectal temperature: There was no significance between groupI and groupII(P>0.05);The rectal temperature decreased at 6h(35.1±0.83oC) after CLP,the lowest was at 24h(33.8±1.45oC),but only the value at 24h was lower significantly than groupI and groupII(P<0.05);The value in groupIV (35.8±0.55oC) was lower than groupIII(36.4±1.05oC) at 3h(P<0.05).4 Mortality: The mortality of groupIII and groupIV was both 50% at 24h after CLP(P>0.05);but it was 75% and 50% respectively at 48h in these 2 groups,and the difference between them was statistically significant (P<0.05);The mortality of other groups and subgroups was 0%.5 E-selectin: The level of serum E-selectin was smallest in groupI and increased slightly in groupII(P>0.05);The level in groupIII increased significantly at 3h after CLP,the peak was at 6h,and showed downward trend after 24h,but they were all higher significantly than groupI(P<0.05) and groupII;The levels in groupIV were all lower respectively than those in groupIII at 3h,6h,24h and 48h,But the difference between the 2 groups was statistically significant only at 3h and 6h(P<0.05).6 vWF: The level of serum vWF was small in groupI and increased slightly in groupII(P>0.05);The levels in groupIII and groupIV were all higher than groupI and groupII (P<0.05);The level in groupIII increased significantly at 3h after CLP,the peak was at 6h,and showed downward trend after 24h,the levels in groupIV were all lower respectively than those in groupIII at 3h,6h, 24h and 48h,the difference between the 2 groups was statistically significant at 3h,6h and 24h(P<0.05).7 iNOS: The iNOS protein of the aorta thoracalis was small in groupI and increased slightly in groupII(P>0.05);The levels in groupIII were higher than groupI and groupII(P<0.05);The level in groupIII increased significantly at 3h after CLP,the peak was at 6h,and showed downward trend after 24h,but still to remain higher level,the values in groupIV were all lower respectively than those in groupIII at 3h,6h,24h and 48h(P<0.05).8 eNOS: The eNOS protein of the aorta thoracalis was the highest in groupI and decreased slightly in groupII(P>0.05);The levels in groupIII were lower than groupI and groupII(P<0.05);The level in groupIII decreased significantly at 3h after CLP,the lowest was at 6h,and showed increasing trend after 24h,the levels in groupIV were all higher than those in groupIII at 3h,6h,24h and 48h,the difference between the 2 groups was all statistically significant except 48h(P<0.05).Conclusions1 A septic model of rat could be made successfully by CLP.2 The level of serum E-selectin and vWF of the septic rats increased significantly.INOS proteinum of the aorta thoracalis increased significantly, while eNOS proteinum decreased significantly.3 Simvastatin could lower the level of serum E-selectin and vWF of septic rats.4 Simvastatin could lower iNOS proteinum of septic rats, but increase eNOS proteinum level.This certified that Simvastatin could regulate the unbalance between iNOS and eNOS at protein level in sepsis,thus it could protect endothelial function.5 Simvastatin could improve the mortality of septic rats at 48h.
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