Study On The Effect Of Locostatin On Intestinal Function In Sepsis Mice

Objective: This paper mainly evaluates the effect and mechanism of Locostatin on the intestinal function of sepsis mice.Methods: Forty mice were randomly divided into Sham + DMSO group,Sham + Locostatin group,CLP + DMSO group,and CLP + Locostatin group.C57 BL / 6 male(n = 40)sepsis model was established by cecal ligation and puncture(CLP)surgery.24 hours,48 hours,and 7 days survival rates were used to study the efficacy of Locostatin in treating sepsis mice.Samples of serum,liver,small intestine,and large intestine of mice in each group were collected on the 7th day after surgery,as well as fecal specimens of mice in each group on the 1st day before and 6th day after surgery,and the changes in body weight of the mice were recorded.Liver function was determined by measuring the levels of ALT and AST in the serum of mice in each group on the 7th day after CLP.Enzyme-linked immunosorbent assay(ELISA)was used to measure the MPO,OPN and E-Selectin levels of liver tissue,small intestine tissue,and large intestine tissue to determine the degree of inflammation.The fecal microbiota diversity of each group of mice was analyzed by high-throughput sequencing 1 day before and 6 days after surgery.Results:1.Survival rate and weight changeThe 24-hour survival rates in the Sham + DMSO group,Sham + Locostatin group,CLP + DMSO group,and CLP + Locostatin group were 100%,100%,80%,and 100%,respectively.The 48-hour survival rates were 100%,100%,50%,and 100%,respectively.The 7-day survival rates were 100%,100%,50%,and 90%,respectively.Within 6 days after surgery,the weight of the mice in the Sham + DMSO group and the Sham + Locostatin group gradually increased,but the weight of the mice in the CLP + DMSO group and the CLP + Locostatin group did not increase significantly.2.Serum ALT and AST levelsCompared with the mice in the Sham + Locostatin group,the serum ALT(P<0.05)and AST(P<0.01)levels of the CLP + Locostatin group increased.Compared with mice in the CLP + DMSO group,the serum ALT content in the mice in the CLP + Locostatin group was increased(P<0.05).3.MPO levels in liver,small intestine,and large intestineCompared with the mice in the Sham + DMSO group,the MPO level in the large intestine tissue in the CLP + DMSO group was significantly increased(P<0.01),and the MPO level in the large intestine tissue in the CLP model group was significantly reduced after Locostatin intervention(P<0.05).4.OPN levels in liver,small intestine,and large intestineCompared with mice in the Sham + Locostatin group and Sham + DMSO group,OPN levels in the large intestine tissue in the CLP + Locostatin group were significantly increased(P<0.01).Compared with the CLP + DMSO group,there was no significant difference in OPN levels in the large intestine tissue of the CLP + Locostatin group(P> 0.05).5.E-Selectin levels in liver,small intestine,and large intestineCompared with the Sham + Locostatin group,the E-Selectin level of liver tissues in the CLP + Locostatin group decreased(P<0.05).Compared with the Sham group,the E-Selectin level in the small intestine of the CLP group was significantly increased(P<0.05).6.Results of diversity analysis of fecal flora in miceThere was no significant difference in the alpha diversity of fecal flora in the Sham + DMSO(SD)group,Sham + Locostatin(SL)group,CLP + DMSO(CD)group,and CLP + Locostatin(CL)group 1 day before surgery(P>0.05).Compared with the mice in the Sham + DMSO group,the alpha diversity of fecal flora in the CLP + DMSO group on the sixth postoperative day was reduced(P<0.05),and the beta diversity was significantly different(P<0.01).Lactobacillus(P-adj <0.05),Proteus(P-adj<0.05),Enterococcaceae(P-adj<0.05),Streptococcus(P-adj<0.05),SMB53 genus(P-adj<0.05),Enterobacteriaceae(P-adj<0.05),Clostridiaceae(P-adj<0.05),Epulopiscium(P-adj<0.05),Ruminococcaceae(P-adj<0.05),Acinetobacter(P-adj<0.05)increased significantly.Bacteroides(P-adj<0.05),Adlercreutzia(P-adj<0.05),Parabacteroides(P-adj<0.05),Oscilillospira(P-adj<0.05)were significantly reduced.Compared with mice in the CLP + DMSO group,there was no significant difference in alpha diversity in fecal flora on the 6th day after surgery in the CLP + Locostatin group(P>0.05),and there was no difference in Beta diversity(P>0.05).But Bacteroides increased significantly(P-adj<0.05).Conclusion: Locostatin can inhibit neutrophil infiltration in the large intestine tissue of mice with sepsis caused by cecal ligation and puncture,reduce the inflammation of large intestine tissue,and increase the abundance of intestinal physiological flora,thereby improving the survival rate of sepsis mice.