| Objective:To observe the effect of Yiqi Wenyang of Shenfu Injection on sepsis rats intervention by detecting the intervention group,model group and Sham operation group of SD rats intestinal epithelial cell tight junction protein Occludin,ZO-1,Claudin-1,Occludin,Claudin-3,the serum tumor necrosis factor(TNF-?),Diamine oxidase(DAO)and D-lactic acid expression and the changes of intestinal epithelial cell ultrastructure.To explore the possible mechanism of Shenfu Injection on rats intestinal mucosal mechanical barrier function and offer the experimental basis for clinical protection of intestinal mucosal mechanical barrier.Method:40 SD rats were randomly selected for Sham group(Sham),sepsis group(CLP),low concentration treatment group(LSF),moderate concentration treatment group(MSF),high concentration treatment group(HSF)by random number table method.The sham group were opened and sutured the abdomen,another rats were caused by cecal ligation and puncture-induced sepsis model of peritonitis.The Sham group and the CLP group were intraperitoneally injected with saline 20ml/kg,while the other three groups were injected 5ml/Kg,10ml/kg(which were diluted with saline to 20ml/kg),20ml/kg intraperitoneal injection with Shenfu Injection.Taking intestinal tissue and serum after 24h,which were used to detect the ultrastructure of intestinal tight junction,intestinal tight junction Occludin,ZO-1,Claudin-1,Claudin-3,serum DAO,D-lactate and TNF-? in expression.Result:1.After 24h,compared with Sham group,the expressions of TNF-?,DAO and D-lactic acid from CLP group rats were 1956.34±51.76p,g/L,3.44±0.14?g/L,124.40±8.87?g/L,which were significantly higher(P<0.05),while the expressions of TNF-? from the LSF,MSF,HSF group were(105.60±2.44)pg/ml,(90.21±5.57)pg/ml and(61.50±7.14)pg/ml,DAO were(1795.79±58.93)U/L,(1171.88±49.1 1)U/L and(915.01±82.45)U/L,D-lactic acid were(2.83±0.06)mmol/1,(2.48±0.09)mmol/1 and(2.45±0.69)mmol/l,which were all P<0.05 compared with CLP.2.After 24h,compared with Sham group,the expression of intestinal epithelial tight junction proteins Occludin,ZO-1,Claudin-1,Claudin-3 from the CLP group were significantly reduced(P<0.05),while the expression from the LSF,MSF,HSF group were 32.91±2.55 and 6.77±1.22,38.93±3.58 and 8.30±2.39,34.73±1.69 and 11.41±1.20,26.65±1.91 and 8.04±1.36,the two groups were(P<0.05),while the expression of Occludin from the LSF,MSF,HSF group were 8.04±1.36,18.35±1.56 and 24.29±9.42,ZO-1 were15.45±2.21,23.22±1.74 and 29.14±2.49,Claudin-1 were 16.06±1.36,20.22±1.13 and 27.04±1.16,Claudin-3 were 18.51 ±0.89 and 22.57±1.03,which were all(P<0.05)compared with CLP.3.The Sham group of ileal mucosa epithelial microvilli arranged neatly,the epithelial tight junctions were continuous and each cell was complete;the CLP group of ileal mucosa microvilli was damaged,the villus subepithelial capillary was congested and shedded,the central lacteal was expanded,the intercellular tight junction was discontinuous,the gap widened,the microvilli became thinner and shorter,and parts fractured,the mitochondrial were severe swelled,the cell organelles damaged and missed lysosomal.The LSF group,MSF group and HSF group rats' ileal epithelium Chiu's scores were all significantly reduced(P<0.05)compared with CLP;and the microvilli arranged in basic rules,tight junctions were continuous.Especially in the HSF group,the intercellular tight junctions were clearly visible.Conclusion:After 5,10,15 ml/kg intraperitoneal injection of Shenfu,abnormal changes of sepsis was significantly improved,including the TNF-?,DAO,D-lactate,Occludin,ZO-1,Claudin-1,Claudin-3 protein expression and the intestinal epithelial cell ultrastructure,and these effects of intensity are dose proportional with Shenfu's. |
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