| Objective:To study the effect and mechanism of "Chai Huang Qing Yi Huoxue Granule" on intestinal mucosal barrier of SAP rats.Method:120 male healthy SD rats were randomly divided into the following 5 groups: sham operation group,SAP model group,low concentration of Chai Huang Qing Yi Huoxue Granule group,medium concentration group,high concentration group,and each group has 24 SD rats.We induced the SAP models by injecting 3.5% taurocholic acid into the pancreatic duct.The successful model rats were anesthetized immediately after awakening,one times about every 4 hours and administrated by 1ml/100 g of the weight.High,medium and low concentration groups were given high,medium and low concentrations of Chai Huang Qing Yi Huoxue Granule separately by gavage method,model group and sham operation group were given saline gavage.At 6h,12 h,24h after modeling,8 rats in each group were anesthetized for laparotomy and were followed up to observe the incidence of hemorrhage and necrosis of pancreas.Prepare serum from abdominal aorta blood to detect hemodlastase,diamine oxidase,d-lactic acid and endotoxin;Having the pancreas and duodenum were stained with HE and observed under light microscope;And immunohistochemical method was used to detect the expression of the tight junction protein Occludin in small intestinal mucosa.Results: 1.SAP rats were successfully modeled: the pelvic hemorrhage and purple-black necrosis were seen in the pancreatic tissue at 6h after modeling.The pancreatic interstitial edema was seen by HE staining,;Blood amylase increased to varying degrees.The above observations are in line with SAP changes,suggesting successful modeling.2.By observing the basic condition of the abdomen we found that in the sap model group there was patchy hemorrhage and purple black necrosis in the pancreatic tissue;In gastrointestinal tract there was obvious gas accumulation,and the wall was thin and translucent;On the greater omentum and the mesentery there was more saponification spot.However,in the high,medium and low concentration group,the pancreas necrosis and the intestinal accumulation of the rats were reduced than that of the model group,and on the greater omentum and the mesentery there was less saponification spot.3.It was reminded that the amylase of the model group,high,medium and low concentration groups was increased in 6,12,24 hours after SAP rats modeling,compared with the sham group(P<0.05).The amylase of high,medium and low concentration groups was decreased in 12 h and 24h(P<0.05);The amylase of medium and low concentration groups was lower than that in the high concentration group in the 12h(P<0.05).The amylase in 24 h from large to small was high concentration group,low concentration group and medium concentration group(P<0.05).4.Through the detection of serum DAO,D-Lactate and endotoxin,we found that serum DAO,D-Lactate and endotoxin in the high,medium and low concentration groups were higher than sham operation group(P<0.05),which showed that the serum DAO,D-Lactate and endotoxin were increased after sap was successfully established.But serum DAO and endotoxin each two compared all have significant difference in the high,medium and low concentration group at 12 hour(P<0.05),D-Lactate in low concentration group at 12 hour has significant difference with the high and medium concentration groups(P<0.05).DAO in low concentration group at 12 hour was higher than that in the model group(P<0.05),and DAO in medium concentration group at 24 hour was higher than that in the model group(P<0.05).Except the DAO in low concentration group was lower than that in the high and medium concentration groups(P<0.05),there are no significant difference between other groups.The results show that Chai Huang Qing Yi Huoxue Granule can reduce the production of the D-Lactate and endotoxin in the SRA rats.The decline from large to small was high concentration group,medium concentration group and low concentration group.5.Observation of pancreatic HE staining under light microscope shows that in the model group the pancreatic tissue was severely damaged and increased with time.And we can see the pancreatic interstitial edema,punctate hemorrhage,inflammatory cell infiltration,focal degeneration and necrosis.The damage of high,medium and low concentrations at 12 h and 24 h was reduced which was compared with the model group.While compared each two in the high,medium and low concentration group at the same time point,the range of the improvement of pancreatic tissue injury from large to small was high concentration group,medium concentration group and low concentration group.Pancreatic pathological score shows that pathological score in high,medium and low concentrations at 24 h has significant differences which compared with the model group,and show an downward trend(P<0.05);the pathological score at 24 h of each two group of high,medium and low concentration groups have differences(P<0.05).The results showed that 24 hours after taking the Huang Qi Qing Yi Huoxue granule could reduce the pathological score of pancreas in SAP rats,and the decline from high to low in order was high concentration group,middle concentration group and low concentration group.Combined with HE staining of pancreatic tissue under light microscopy and pathological score,the results showed that the amylase increased after the success of the model of SAP rats,but the degree of increase was not significantly associated with the severity of pancreatitis.6.Through the observation of small intestinal mucosa under light microscope we found that the intestinal mucosa in the model group was severely damaged and increased with time,and congestion and edema of intestinal villi with hemorrhage,massive neutrophil infiltration,reduction of goblet cell and apical epithelial necrosis,shedding,ulceration can be seen.The injury of the high,medium and low concentration groups at 12 h and 24 h were alleviated compared with the model group.But,the high,medium and low concentration group were compared with model group of the same time points,with the time increase congestion and edema of intestinal villi,necrosis and inflammatory cell infiltration were improved,suggesting that Chai Huang Qing Yi Huoxue granule can reduce the sap of intestinal mucosa injury in rat,and the mitigation degree of intestinal mucosal injury from high to low was high concentration group,medium concentration group and low concentration group.7.The expression of Occludin in small intestinal mucosa of rats was studied,it pointed out that the expression of cell membrane and membrane in the model group,the high,medium and low concentration groups were higher than that in the sham operation group.The results showed that the expression of Occludin in intestinal mucosa was decreased after the successful modeling of SPA.But compared with the model group at the same time point,the expression of Occludin of the high,medium and low concentration groups were gradually increased,the expression was enhanced,and gradually increased with time.The results indicated that the Chai Huang Qing Yi Huoxue granule could improve the expression of tight junction protein Occludin in SAP rats,and the degree of recovery from high to low was high concentration group,medium concentration group and low concentration group.Conclusion:SAP damage can leads to elevated concentration of serum amylase,D-lactate,DAO and plasma endotoxin,and lead to the expression of tight junction protein Occludin was decreased in small intestinal mucosa.Early application of compound preparation of traditional Chinese medicine of Chai Huang Qing Yi Huoxue granule can improve SAP rat's pancreas,intestinal mucosal injury,reduce serum D-lactic acid,DAO,plasma endotoxin,improve the expression level of intestinal mucosal tight junction protein Occludin,and improve the function of intestinal mucosa barrier. |
PDF Full Text Request