| Objective: SAP rat model was establish through the detection of related blood indicators and pathological observation, which was aimed to have an understanding of the changes of intestinal permeability and intestinal permeability in SAP rats; to observe the expression of NF-κB, TLR4 and ZO-1 in the intestinal mucosa of SAP rats; to investigate the protective effect and mechanism of traditional Chinese medicine Qingyi decoction on gut barrier function in SAP rats. Methods: 54 SD rats at Clean Level, male, weighing 280 + 20 g. randomly divided into three groups: sham operation group(close the abdominal after its entering the abdomen, only flipping pancreas several times), SAP group(5% sodium taurocholate SAP model) and Qingyi Decoction treatment group(Immediately after the SAP model was established, Qingyi decoction was administered to the stomach, 1ml/100g/ time, once per 12h). Each group was tested with three different lengths of time: 12 h, 24 h, 48 h,with 6 rats at each point of time. Blood, pancreatic tissue and intestinal tissue samples were taken at different time points in each group. Blood samples were used to detect the levels of serum amylase, endotoxin, cytokines TNF-, IL-6 and D-lactic acid. Pancreatic tissue and intestinal tissue were respectively used for HE staining and immunohistochemical staining, and then the pathological changes of pancreatic and intestinal tissues and the expression of NF-κB, TLR4 and ZO-1 in small intestinal mucosa were observed. Results:(1) The level of serum amylase in SAP group was significantly higher than that in SO group at different time points(P<0.05);the level of serum amylase in Qingyi decoction group was significantly lower than that in SAP group at all time points, and higher than that in SO group(P<0.05).(2) The levels of endotoxin in SAP group and Qingyi decoction group were significantly higher than that in SO group at different time points(P<0.05);Qingyi decoction group were lower than the SAP group at all time points, and statistical significance emerged in the difference at the points of 12 h and 48h(P<0.05). The endotoxin levels in SAP group at the time point of 24 h were significantly higher than those at the time points of 12 h and 48h(3) Similarly, IL-6 and TNF-α in Group SAP reached the top at 24 h, and, comparatively, they were distinctly higher than those in Group SO at 12 h, 24 h and 48h(P<0.05); IL-6 and TNF-α in Qingyi decoction group were apparently lower compared with those in Group SAP, while they were obviously higher than those in Group SO, and the statistical significance in the difference was seen at both 12 h and 48h(P<0.05).(4) D-lactic acid levels in SAP group were significantly higher than those in SO group at two time points, 12 h and 48h(P<0.05);D-lactic acid levels in Qingyi decoction group were significantly lower than those in SAP group and higher than the SO group at two time points, 12 h and 48h(P<0.05). The D-lactic acid level of SAP group was the highest at the time point of 12 h, which decreases gradually with the passing of time(P<0.05).(5) Pancreatic tissue pathological score in group SAP was significantly higher than that in SO group at all time points(P<0.05);Pancreatic tissue pathological score in Qingyi decoction group was significantly lower than that in SAP group and was higher than that in SO group at all time points(P<0.05).(6) The histological grade of SAP group was significantly higher than that in SO group at all time points(P<0.05); The histological grade of Qingyi decoction group was lower than that of SAP group and higher than that of SO group at each time point(P<0.05).(7) In SAP group and Qingyi decoction group the expression of NF-κB P65 in intestinal mucosa was higher than that in SO group at each time point, and there was a significant difference at the time point of 12h(P<0.05); The expression of NF-κB P65 in the intestinal mucosa epithelial cells in the Qingyi decoction group was lower than that in the SAP group and higher than the SO group at the time point of 12h(P<0.05). The expression of NF-κB P65 in SAP group was distinctly higher at the time point of 12 h than at 24 h and 48h(P<0.05).(8) The expression of intestinal epithelial TLR4 in SAP group and Qingyi decoction group was distinctly higher than that in SO group at all time points(P<0.05); At the time point of 12 h,the expression of intestinal epithelial TLR4 in the Qingyi decoction group was apparently higher than that in the SO group and lower than that in the SAP group(P<0.05); The expression of intestinal epithelial TLR4 in SAP group reached the peak at the time point of 12 h,and decreased with the increasing of time(P<0.05).(9) The expression of ZO-1 in the intestinal mucosa of SAP group and the Qingyi decoction group was significantly lower than that in SO group at all time points(P<0.05);At each time point Qingyi decoction group was distinctly lower than the SAP group and higher than the SO group(P<0.05). Conclusion:(1) Intestinal pathological changes and damages to the intestinal barrier function happened to the SAP rats in the early stage.(2) The expression of TLR4 and NF-κB in the intestinal mucosa of SAP rats is distinctly up-regulated, while the expression of ZO-1 is apparently down-regulated.(3) The Qingyi decoction can apparently reduce the damage of pancreas and small intestine, reduce intestinal bacteria and endotoxin translocation, reduce the release of inflammatory factors, correct the destruction of the intestinal mucosa and protect the intestinal barrier function of SAP. The protective effect of Qingyi decoction may be related to the fact that it inhibits the expression of TLR4/NF- kappa B signaling pathway in intestinal mucosal cells and promotes the expression of ZO-1 in intestinal mucosa cells. |
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