Effects Of Early Bundle Therapy On Intestinal Micro-ecology Of SD Rats Of Sepsis

Objective To investigate the effects of the early bundle therapy on the intestinal microecology of sepsis rats,and the correlation between changes in body and intestinal immune function in rats with sepsis,and to reveal the efficacy of the early bundle in the treatment of sepsis.The mechanism provides a theoretical basis for elucidating the mechanism of development of sepsis and lays a foundation for the exploration of sepsis diagnosis and optimization of treatment protocols.Materials and methods Eighty male rats of SD(200-250g)were randomly divided into 5groups: sham group,sepsis group(CLP),the early bundle therapy group(imipenem+NS group),cyclophosphamide group(CTX),the early bundle therapy combined with cyclophosphamide group(imipenem+NS+CTX group).The sepsis model of SDs was replicated by cecal ligation and puncture(CLP).The sham group was given the same procedure but no perforation and ligation.After the success of the sepsis model was judged and identified,it was treated by intraperitoneal injection of physiological saline(35 ml/kg),imipenem(120 mg/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 by HE to observe the intestinal tissue damage in each group.The apoptosis of intestinal mucosal cells was qualitatively and quantitatively analyzed by TUNEL method,and the change of intestinal permeability was further analyzed.Western blot technique was used to detect the expression of occludin in colon tissues of rats in each group,and the changes of intercellular permeability were analyzed.By ELISA,the expression levels of inflammatory cytokines(IL-6,IL-10,TNF-α)were detected and the effect of inflammatory factors on intestinal permeability was analyzed.Results 1.The sepsis model was successfully replicated;2.The 7-day survival rate of each group was observed.The survival rates of CLP group,imipenem+NS group,and imipenem+NS+CTX group were 0%,50%,and 70% respectively.The imipenem+NS+CTX group can significantly improve the survival rate of SD rats.The difference was statistically significant(p<0.05);3.Pathological lesions of the intestinal mucosal injury were observed by pathological section HE staining,and TUNEL was used to detect the apoptosis of intestinal epithelial cells.The results showed that Intestinal mucosal injury and epithelial cell apoptosis were most obvious in the CLP group;Intestinal mucosal injury and intestinal epithelial cell apoptosis were improved in the other treatment groups to varying degrees.In the CLP group,intestinal epithelial cell apoptosis and imipenem+NS group Compared withimipenem+NS+CTX group,the difference between NS group and imipenem+NS+CTX group was statistically significant(p<0.05);4.Western-blot technique was used to detect the expression of occlucin protein in intestinal tissue,and fluorescence colorimetry was used to detect plasma FD-70 level.Changes in intestinal permeability.The results showed that the expression of occludin protein was significantly down-regulated in the sepsis group,the plasma FD-70 level was significantly increased,and the intestinal permeability was significantly increased.The occludin protein expression levels in the imipenem+NS group and the imipenem+NS+CTX group were both with varying degrees of up-regulation,plasma FD-70 levels and intestinal permeability were significantly reduced.The difference was statistically significant(p<0.05);5.Inflammatory factors can reflect the body’s immune function,but also can affect the intestinal wall tissue permeability.The results showed that plasma levels of TNF-α and IL-6 were significantly increased in the CLP group,and plasma levels of TNF-α and IL-6 decreased in the imipenem+NS group and the imipenem+NS+CTX group.The plasma level of IL-10 in the CLP group and imipenem +NS group were significantly higher than that in the sham group,and the plasma level of IL-10 in the imipenem+NS+CTX group was decreased.The difference was statistically significant(p<0.05).Conclusion1.The early bundle therapy can improve the early survival rate of rats with sepsis.The early bundle therapy combined with low-dose cyclophosphamide significantly improved the survival rate of sepsis rats.2.The early bundle therapy has protective effects on the injury of intestinal mucosal barrier in sepsis rats.The early bundle therapy combined with low-dose cyclophosphamide can effectively reduce the damage of intestinal mucosal barrier in sepsis rats,which is related to the changes of intestinal immune function in sepsis rats.