Effects Of Early Liquid Resuscitation On Intestinal Microecology In Rats With Severe Sepsis

Objective : To investigate the effects of early fluid resuscitation therapy and early fluid resuscitation combined with immunomodulatory therapy on the intestinal microecology of severe sepsis rats,and to reveal the therapeutic mechanism of early fluid resuscitation and early fluid resuscitation combined with immunomodulatory therapy for severe sepsis.And the mechanism of the intestinal microecology of rats with severe sepsis provides a theoretical basis for elucidating the mechanism of occurrence and development of severe sepsis,and lays the foundation for the exploration of the diagnosis and treatment of severe sepsis.Methods:Eighty male SD rats(200-250g)were randomly divided into 5 groups: sham group,severe sepsis group(CLP),liquid resuscitation group(NS),cyclophosphamide group(CTX),fluid resuscitation Combined cyclophosphamide group(NS+CTX).The severe sepsis model was replicated by cecal ligation and puncture(CLP).Sham group was given the same procedure but no perforation and ligation were performed.After judging and identifying the successful replication of the severe sepsis model,it was treated by intraperitoneal injection of physiological saline(30 ml/kg)and cyclophosphamide(10 mg/kg).The survival time and number of rats in each group were recorded and the survival curve was drawn.The survival rate of each group of rats was quantitatively analyzed.The concentration of FD-70 in portal vein blood of rats in each group was detected by fluorescent tracer method,and the intestinal permeability of rats in each group was quantitatively analyzed.The jejunum tissue sections of rats in each group were stained with HE to observe the intestinal tissue damage in each group.The TUNEL method was used to qualitatively and quantitatively analyze the apoptosis of intestinal mucosal cells in each group of rats.The rat intestine was further analyzed and analyzed.Cross-cell permeability changes.Western blotting technique was used to detect the expression of occludin in colon tissues of rats in each group,and the changes of paracellular permeability were investigated.The expression levels of cytokines(IL-6,IL-10,TNF-?)in each group were detected by ELISA,and the effect of inflammatory factors on intestinal permeability in rats was analyzed.Results:(1)A rat model of severe sepsis was successfully replicated.The 7-day survival rate of rats with severe sepsis was zero.After cyclophosphamide,liquid resuscitation combined with cyclophosphamide treatment,the survival rate of severe sepsis rats was significantly increased,respectively 10% and 30%.(2)Detection of FD-70 level in portal vein blood and quantitative analysis of changes in intestinal permeability.Intestinal permeability was significantly increased in the CLP group.Intestinal permeability was observed in severe sepsis rats in each treatment group.Significantly improved,while CTX group,NS group and NS+ CTX group,NS + CTX group rats with severe sepsis intestinal permeability was significantly lower than NS group(P <0.05)and CTX group(P <0.01)There was no significant difference between the NS group and the CTX group.(3)Intestinal tissue sections were stained with HE and TUNEL assay was used to detect the number of apoptotic cells.Pathological lesions of intestinal tissues and intestinal epithelial cell apoptosis were detected,and the extent of intestinal cell damage and changes in transmembrane permeability were evaluated.The results showed that there were obvious intestinal tissue damage and apoptosis in the CLP group,while the intestinal tissue damage was significantly improved in the CTX group,the NS group,and the NS+CTX group,and the apoptosis of the intestinal epithelial cells was significantly reduced.(4)Quantitative detection of occludin expression was used to investigate the effect of paracellular permeability.The results showed that the expression of occludin was significantly reduced in CLP group,and occludin was expressed in CTX group,NS group and NS+CTX group.The amount increased(P<0.05),while there was no significant difference between the CTX group and the NS+CTX group.(5)The expression levels of inflammatory factors in the body showed that the expression levels of IL-6,IL-10 and TNF-? in the peripheral blood were significantly higher in the CLP group than in the intervention group(P<0.01);(IL-6,TNF-?): Expression levels in each treatment group decreased(P<0.05),whereas expression levels in NS+CTX group decreased significantly(P<0.01).Anti-inflammatory cytokines(IL-10): CTX group Compared with CLP group,NS group and NS+CTX group,the expression level in peripheral blood decreased(P<0.05),while the expression level in NS+CTX group increased significantly(P<0.01).Conclusion:(1)Early fluid resuscitation treatment can improve the early survival rate of rats with severe sepsis;but it can not reduce the mortality rate of severe severe septic rats.Early fluid resuscitation combined with low-dose cyclophosphamide can significantly improve the survival rate of rats with severe sepsis.(2)Early fluid resuscitation has a protective effect on intestinal mucosal barrier injury in rats with severe sepsis.Early fluid resuscitation combined with low-dose cyclophosphamide can effectively reduce the damage of intestinal mucosal barrier in severe sepsis rats,which is related to the changes of intestinal immune function in severe sepsis rats.