The Research Of Differential Proteins In Ileum Tissue By Probiotic In Response To Severe Acute Pancreatitis

Objective: Through the establishment of the model of severe pancreatitis rat,.Comparing the differential expression protein in the ileum of rat between severe acute pancreatitis group,probiotic intervention group and sham operation group,providing the basis for the study of the intestinal mucosa barrier damage when SAP occurs by the Screening of specific expression,providing also the new theoretical basis and protein molecular targets for the probiotic in the hands of remedy and prevention of he intestinal mucosa barrier damage in severe pancreatitis rat,Methods:(1)8 weeks 54 healthy male SD rats,Weighting 250 ~ 300 g,Randomly divided into 3 groups,;sham operation group(SO group,n= 18)?severe pancreatitis group(SAP group,n = 18)?Probiotics treatment group(YSJ group,n = 18).(2)opening SO group abdominal cavity ?turning of the pancreas and closed it.Establishing SAP animal model in rats by 3% sodium taurocholic acid in SAP group.Establishing SAP animal model in rats by 3% sodium taurocholic acid in YSJ group,filling the stomach of SO rats with the 10% probiotic solution,a total of 2 times;filling the stomach of SAP group rats with the normal saline,a total of 2times;(3)The three groups of rats in the pancreas,kill animals 24 h time points after take the specimen of the ileum tissue and rat abdominal venous blood.(4)Pancreas and the ileum organization HE staining,grading the pancreas by using kusske standard line of pathological grading,grading the ileum organizations by using improved Chiu's score method.(5)Each ileum tissue samples,extracting protein,denaturating protein,reduction and enzyme solution.Then iTRAQ labeling on the upgrade reverse dphase liquidchromatography-tandem mass spectrometry(nanoRPLC-MS/MS)technology for each protein,identification and bioinformatics analysis of samples.(6)The data analysis;The collected Data by using SPSS17.0 statistical software for statistical analysis,the sample mean is using single factor analysis of variance,P< 0.05 be thought the difference was statistically significant,Differences in protein screening by R is calculated using the t test software to its significant index(pValue),respectively,using p-Value < 0.05 and a Fold change>1.5 or a Fold change<0.6667 protein screening.Results:(1)3%STC had successfully induced SAP rat model,the pancreas had a typical pathological change.(2)The pancreatic pathology score Of SAP and YSJ group were higher than sham operation group(SO group)(P?0.05),YSJ pancreatic pathology score was higher than SAP group(P ? 0.05).3%STC had successfully induced SAP rat model,the pancreas had a typical pathological change.(3)The ileum pathology score of SAP and YSJ group were higher than sham operation group(SO group)(P?0.05),YSJ pancreatic pathology score was higher than SAP group(P?0.05),with statistical significance(P<0.05).YSJ pathological score lower than SAPgroup,the difference was statistically significant(P < 0.05);(4)The D-lactic acid and endotoxin of SAP group were higher than YSJ group were higher than control group(SO)(P?0.05),the D-lactic acid and endotoxin of YSJ group were lower than SAP group(P?0.05).(5)the SAP group and SO screening a total of 37 significant differential proteins,6 proteins up-regulate,31 proteins down-regulate;YSJ screened 94 differences between groups and SO protein,6 proteins up-regulate,88 proteins down-regulate;YSJ with SAP group have in common 7 proteins,which are zonule protein 1(ZO-1),Intestinal mucin-like protein,Chaperonin 10,Cytosolic phospholipase A2 ? Desmin,Intelectin1,Transgelin.(7)immunohistochemical validation showed that the expression ZO-1protein in the SO group was down-regulated in SAP group(P<0.05),while YSJ group expression level in the SAP group.Conclusion:The research of differential in protein based on iTRAQ technology provides the reference for the study of the intestinal mucosa barrier damage mechanism for SAP,also can well reflect the phenomenon of probiotics in SAP rats ileum proteins.The differential proteins may provide new theoretical basis for probiotics treatment of SAP may also be the intervention therapeutic targets.