| Objective: By establishing the animal model of severe acute pancreatitis(SAP)with intestinal failure induced by sodium taurocholate in rats,the changes of serum motilin,gastrin,diamine oxidase and d-lactic acid in rats were observed,the effects of bifidobacterium quadruple live bacteria tetrad on intestinal mucosal barrier function in SAP rats and the possible mechanism were discussed,and the preventive and therapeutic effects of bifidobacterium quadruple live bacteria tetrad on intestinal dysfunction in SAP animal model were evaluated,It provides a theoretical basis for micro-environmental nutrition treatment for SAP patients with intestinal failure.Methods:(1)Twenty-four male Wistar rats with body weight of 200-250 g were randomly divided into three groups: probiotics group,model group and blank group,with 8rats in each group.(2)The rats in the probiotics group were given 0.75 g BID of bifidobacterium quadruple live bacteria 7 days before to 24 hours after model establishment,and the rats in the model group were given 0.75 g BID of corn chips 7 days before to 24 hours after model establishment.The rats in the probiotics group and the model group were slowly injected 3.5% sodium taurocholate through the pancreaticobiliary duct to make the SAP model of rats.(3)After 24 hours of modeling,the animals were anesthetized and the samples were taken.Five ml of blood was taken from the heart of the rats.The whole blood was taken to detect the white blood cell count.After centrifugation,the serum was taken to detect the levels of c-reactive protein and amylase.The levels of serum motilin,gastrin,diamine oxidase and d-lactic acid were measured by ELISA.The pancreatic gland tissue was taken for pathological section and HE staining to determine whether SAP modeling was successful and to observe the degree of pancreatic injury.(4)The data collected above are statistically processed with SPSS 27.0 software.Measurement data conforming to the normal distribution were expressed using(x ± s),inter group comparisons were performed using one-way analysis of variance,and pairwise comparisons were performed using the least significant difference method(LSD method);Measurement data that do not conform to the normal distribution are represented by M(P25,P75),and comparisons between the two groups are performed using nonparametric rank sum tests.When P<0.05,the difference of data is considered statistically significant.Results:(1)Pathological results of pancreas: In the blank group,the structure of pancreatic lobule is complete,the interlobular space is normal,the acini,ducts and islets are clearly visible,the cells are closely arranged,and the acinar cells can be seen with punctate nuclei,without hemorrhage or necrosis.In the model group,the structure of the pancreatic lobule was blurred,the lobule disappeared,the lobule space was obviously swollen and widened,the nucleus of some acini was fragmented or dissolved,and a large amount of coagulative necrosis appeared.Inflammatory cells such as monocytes and neutrophils could be seen around the necrotic pancreatic tissue,and some areas had fat liquefaction necrosis,interstitial small blood vessel wall bleeding and necrosis,and a large number of red blood cells overflowed.In the probiotics group,the pancreatic tissue structure is generally visible,the lobular structure is slightly disordered,the lobular space is widened,and the degree of edema is lighter than that in the control group.The acinar cells are swollen,and the swollen nuclei are still visible.Some pancreatic cells are necrotic,without fat liquefaction necrosis,and the infiltration of inflammatory cells is significantly weaker than that in the model group.(2)The white blood cell count in the probiotics group and the model group was significantly higher than that in the blank group,the difference was statistically significant(P<0.01);Compared with the model group,the white blood cell count in the probiotics group was significantly decreased(P<0.01).(3)The concentration of c-reactive protein in probiotics group and model group was significantly higher than that in blank group(P<0.01);Compared with the model group,the concentration of c-reactive protein in the probiotics group was significantly decreased(P<0.01).(4)Compared with blank group,serum amylase concentration in model group was significantly increased(P<0.01);Compared with the model group,the serum amylase concentration in the probiotics group was significantly decreased,and the difference was statistically significant(P<0.01).(5)Serum motilin concentration in model group was significantly decreased compared with blank group(P<0.01);Compared with the model group,the serum motilin concentration in the probiotics group was significantly increased(P<0.05).(6)Compared with blank group,serum gastrin concentration in model group was significantly increased(P<0.05);Compared with the model group,the serum gastrin concentration in the probiotics group was significantly decreased(P<0.05).(7)Compared with blank group,serum diamine oxidase concentration in probiotics group and model group was significantly increased(P<0.05);Compared with the model group,the serum diamine oxidase concentration in the probiotics group was significantly decreased.(8)The concentration of d-lactic acid in probiotic group and model group was significantly higher than that in blank group,and the difference was statistically significant(P<0.01);Compared with model group,the concentration of d-lactic acid in probiotics group was significantly decreased(P<0.01).Conclusion:(1)Bifidobacterium quadruple live bacteria can alleviate pathological injury and inflammatory reaction in SAP rats,and play a role in protecting pancreatic tissue.(2)The MLT concentration of the probiotics group was higher than that of the model group,and the GAS concentration was lower than that of the model group.Considering that bifidobacterium quadruple live bacteria tetrad could improve the gastrointestinal function of SAP rats,its mechanism was related to regulating the secretion of MLT and GAS.(3)The DAO and D-LA concentration in the probiotics group was lower than that in the modell group,so it was considered that bifidobacterium quadruple live bacteria tetrad can protect the intestinal barrier function of SAP rats. |
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