| Objective Previous basic experimental studies have found that exogenous citrulline can inhibit the release of proinflammatory factors in early sepsis by regulating Cit-NO cycle and JAK-STAT signaling pathway,and reduce the degree of inflammation in important organs such as heart,liver and kidney.On the basis of this study,we further explored the effects of citrulline on ileal mucosal histology and plasma levels of diamine oxidase and D-lactic acid in sepsis rats.Methods The sepsis rat model of cecal ligation and perforation(CLP)was established.SD rats were randomly divided into normal group,citrulline gastric lavage group,sham operation group,simple CLP operation group,citrulline gastric lavage + CLP operation group.Normal group was fed routinely without any special treatment.Citrulline solution was given to rats in citrulline gastric lavage group at a dose of 200 mg/kg*d per day for 10 consecutive days.Rats in sham operation group underwent sham operation and opened abdominal cavity under chloral hydrate anesthesia.Rats in the CLP group underwent cecal ligation and perforation,while rats in the citrulline plus CLP group were given citrulline solution at a dose of 200 mg/kg*d per day for 10 consecutive days before cecal ligation and perforation.The rats in the CLP model group and the citrulline plus CLP operation group were sampled at 6h,12 h,24h and 48 h after the establishment of the model,while the rats in the normal control group,citrulline gastric lavage group and sham operation group were sampled and executed 24 hours after the establishment of the model group.Before sampling,the general appearance,activity,dietary status and abdominal lesions of rats were observed.Rat arterial blood was collected by abdominal aorta blood sampling method and serum was separated at room temperature.The expression levels of TNF-a,DAO and D-lactate in plasma were detected by ELISA kit.Each rat took a around 2 cm long section of ileal tissue from the ileocecal part about 10 cm away,fixed with formaldehyde and embedded with paraffin,and then stained with HE.Pathological changes of ileal tissue and mucosa were observed under pathological microscope and pathological grade of ileal mucosa injury was scored.SPSS 20.0 statistical software package was used for analysis.The difference was statistically significant between the two sides(P < 0.05).Results 1.Normal group,citrulline gastric lavage group and sham-operated group were in good condition before sampling,and there was no obvious inflammatory lesion in abdominal cavity.Rats in the model group of simple cecum ligation and perforation operation gradually showed mental depression,tremor and loss of appetite after operation.With the passage of time,intestinal canal dilatation,congestion and edema,hemorrhagic exudation and necrosis appeared in varying degrees.The general condition and intraperitoneal inflammatory lesions of rats in the citrulline plus CLP group at different time points after operation were improved compared with those in the CLP group alone.2.Compared with the normal group,citrulline gastric lavage group and sham operation group,mucosal edema,inflammatory cell infiltration and villus exfoliation occurred in ileum mucosa of CLP group and citrulline gastric lavage + CLP group over time.At 6 hours after operation,there was no significant difference in the pathological damage score of ileal mucosa between the citrulline plus CLP group and the simple CLP group(P > 0.05),but at 12,24 and 48 hours after operation,the pathological damage score of ileal mucosa was significantly lower than that of the simple CLP group(P < 0.05).3.Compared with the normal group,citrulline gastric lavage group and sham-operation group,the serum TNF-alpha levels of CLP group and citrulline gastric lavage + CLP group began to rise at about 6 hours after operation,then reached the peak at about 12 hours after operation,then declined,remained at a high level at 24 hours after operation,and gradually decreased to the initial state 48 hours after operation,and citrulline was in the initial state.The average level of serum TNF-alpha in the group of intragastric administration plus CLP at 6,12 and 24 hours after operation was lower than that in the group of simple CLP operation(P < 0.05),but there was no significant difference between the two groups at 48 hours after operation(P > 0.05).4.Compared with the normal group,citrulline gastric lavage group and sham-operation group,the levels of serum diamine oxidase in CLP group and citrulline gastric lavage + CLP group increased with time.There was no significant difference in the levels of serum diamine oxidase between the citrulline gastric lavage + CLP group and CLP group at 6 hours after operation(P > 0.05).The levels of diamine oxidase and D-lactate in serum at 12,24 and 48 hours after operation were lower than those in CLP group(P < 0.05).5.Compared with the normal group,citrulline gastric lavage group and sham-operated group,the serum D-lactic acid levels in the CLP group and the citrulline gastric lavage + CLP group increased with time.Compared with the CLP group,the serum D-lactic acid levels in the citrulline gastric lavage + CLP group had no significant difference at 6 hours after operation(P > 0.05),but at three time points of 12 hours ? 24 hours and 48 hours after operation,The expression of D-lactate in serum was lower than that in CLP group(P < 0.05).Conclusion The development of sepsis in rats is accompanied by intestinal mucosal damage and increased permeability.Supplementation of exogenous citrulline can alleviate the inflammatory reaction of intestinal mucosa and reduce the permeability of damaged intestinal mucosa,thus playing a protective role on intestinal mucosa. |
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