| Objective:Studying the effect of anti-infection treatment and anti-infection combined immunity treatment in septic rats on intestinal microecological.Revealing the anti-infection treatment and anti-infection combined immunity treatment in septic rats on mechanisms of the curative effect of treatment,and the influence on sepsis in rats intestinal microecological mechanism,to clarify the development mechanisms of sepsis provide theoretical basis,to explore the diagnosis and treatment of sepsis laying a foundation for optimization.Methods : 66 150-250 g SD male rats were randomly divided into 5 groups(sham operation,sepsis,cyclophosphamide,imipenem,and imipenem combined cyclophosphamide).Sepsis model was replicated by caecal ligation and puncture [CLP],and the sham operation group performed the same operation without perforation and ligation.In addition,50 rats were randomly selected for survival analysis using the same model and grouping(10 rats in each group).The treatment was administered by intraperitoneal injection of imipenem(120 mg/kg)and cyclophosphamide(10 mg/kg)and the drug was administered every 12 hours after induced sepsis.Survival analysis animal was continuous treated 7 days,recorded the survival time and number of rats in each group and plotted 7 days survival curve,quantitative analysis of the survival data of each group.The function analysis and mechanisms of intestinal barrier were explored after 24 hours induced sepsis.The FD-70 concentration in rats was detected by fluorescent tracer method,and the intestinal permeability of rats was quantitatively analyzed.The intestinal tissue injury in septic rats in each group was observed through HE tissue section,and TUNEL method was used to characterize the apoptosis of intestinal mucosa cells.The western blot was used to detect the expression level of tight junction protein(occludin)in the intestine,and to explore the analysis of the permeability changes of the cells.The expression levels of cytokines(IL-6,IL-10,TNF-α)were measured by enzyme-linked immunosorbent assay [ELISA] method,and the effect of cytokines on intestinal permeability was analyzed.Results: 1)The sepsis model was successfully replicated with a 7 day survival rate of 18.18%.The survival rate was significantly increased(50% vs 18.18%)in the sepsis group,while the survival rate of sepsis rats in the combined treatment group was improved(70% vs 50%).(2)Intestinal permeability was quantitatively analyzed by explored FD-70 expression change in plasma.Intestinal permeability increased obviously in septic rats,imipenem treatment and combination groups of septic rats’ permeability improved significantly,and imipenem group compared with the combined treatment group,combination group intestinal permeability is higher than the imipenem group.(3)Further explore possible mechanisms of increased permeability in combination therapy group.HE pathological was used for qualitatively detecting intestinal tissue biopsy.TUNEL was used for qualitatively detecting intestinal mucosa cells apoptosis analyzed intestinal cellular permeability.(4)Quantitative detection of the expression level of the intestinal tight junction protein(occludin)was used to further explore the influence of the analysis of the paracellularpermeability.The expression of occludin protein on intestinal tissue in septic rats decreased,imipenem and combined treatment groups’ occludin expression on intestinal tissue quantity increases.The expression of occludin protein in combined treatment group was obviously lower imipenem treatment group.The effects of cytokines on intestinal permeability were further discussed.Results show that the sepsis group of IL-6,IL-10,the TNF-α expression are significantly increased,IL-6,IL-10,the expression of TNF-α are significantly reduced in imipenem group,and the IL-6,IL-10 express further reduced,and the expression of TNF-α increased in combined treatment group.Conclusion:(1)Imipenem significantly improved the survival rate of rats with cecum perforation sepsis;Imipenem has protective effect on intestinal mucosal barrier injury in rats with sepsis.(2)Imipenem combined with low dose cyclophosphamide treatment significantly improved the survival rate of sepsis rats compared with the imipenem alone;Imipenem combined with low dose cyclophosphamide cause aggravating intestinal mucosa barrier dysfunction in septic rats,its possible mechanisms are IL-10 factor and occludin protein expression. |
