| Objective Our previous study showed that supplementation of exogenous citrulline could protect the liver,heart and kidney of septic rats and effectively inhibit the release of early inflammatory factors.Based on this research,we further explore the protective effect of citrulline on the colon and high mobility group box-1 protein（HMGB1）of rats with cecal ligation and puncture sepsis,in order to further explore the possible mechanism of citrulline on sepsis.MethodsThe cecal ligation and puncture（CLP）model was used in our study.Rats were divided into Normal group,NS group,Cit group,sham group,CLP group and CLP+Cit group.Group Normal rats do not do any special treatment.Rats in group NS need to use normal saline for a week of 5ml per day.Group Cit rats need continuous use of citrulline for one week respectively according to the dosage of each 200mg kg-1·d-1.The rats in group Sham were treated with a sham operation,that is,the abdominal cavity was opened after the intraperitoneal injection of chloral hydrate without CLP,and the incision was sutured immediately after the cecal intestinal tube was slightly moved.Rats in group CLP were operated on by CLP.Each CLP+Cit group rat was given preoperative citrulline continuous intragastric administration for one week at a dose of 200 mg/（kg·d）.The observation time for CLP group and CLP+Cit group were 6 h,12 h,24 h and 48 h after CLP.The survival time of rats was recorded at the time of sampling,and the general condition and abdominal lesion were observed before the sampling.The rat abdominal aorta blood was collected and the serum was separated.Detection of HMGB1 level in rat serum by ELISA Kit.The non necrotic colonic tissues of each group were collected,paraffin embedded and sectioned,observation of pathological changes of colon tissue by HE staining.Using SPSS 17.0 for statistical analysis,P<0.05,bilateral difference was statistically significant.Results When 6h and 12h opened the abdominal cavity of group CLP and group CLP+Cit,the abdominal cavity could be observed in the abdominal cavity of the two groups,but there was no difference between the two groups.The symptoms of sepsis in the abdominal cavity of the group of CLP group were more severe than that of the CLP+Cit group at 24 h and 48 h,and more obvious at 24 h.Colonic histopathology and HE staining showed that the infiltration of inflammatory cells in colon tissue of CLP+Cit group was lighter than that in CLP group.The survival rate curves of four time points within 48h of group CLP and group CLP+Cit were compared,and there was a statistical difference between two survival rates at the time point of 48h（P<0.05）.It was considered that supplementation of exogenous citrullinate at 48h could effectively improve the survival rate of sepsis rats.The serum HMGB1 level of rats in CLP group and CLP+Cit group increased gradually before 24h,and the highest value was measured at 24 h and decreased significantly at 48h.At the three sampling time points,the average level of HMGB1 measured in CLP+Cit group was lower than that in CLP group,and at the four time points of 12h,24h and 48h,the group was significantly lower than that of the group（P<0.05）.ConclusionSupplementation of exogenous citrullinate can effectively relieve the symptoms of intraperitoneal inflammation in septic rats,increase the cumulative survival rate at 48h of sepsis rats,alleviate colonic tissue inflammation in septic rats,protect colon tissue and inhibit the release of serum HMGB1 in rats with sepsis. |
