磷酸化蛋白激酶B/蛋白激酶B在不同时间大鼠重症急性胰腺炎肠道损伤的表达

摘要

目的 用5％牛磺胆酸钠诱导大鼠重症急性胰腺炎(SAP)模型,观察磷酸化蛋白激酶B(p-Akt)在其中的表达,并探讨制作该模型合适的时间点.方法 雄性SD大鼠59只,随机分为5组:正常对照组(n=12)、假手术组(CON组,n=12)、SAP组(n=36)、3、6、12 h组(n=12).胆胰管逆行注射5％牛磺胆酸钠制备SAP肠道损伤模型.在3、6、12h测定腹水量,血清淀粉酶、脂肪酶水平,光镜观察胰腺及回肠病理改变,免疫组织化学观察末端回肠蛋白激酶B(Akt)及p-Akt表达.结果 SAP组随着时间点的延长,血清淀粉酶、脂肪酶水平,胰腺和回肠的病理评分逐渐升高,并且p-Akt在SAP大鼠回肠中表达也逐渐增强,在12h达到高峰(P＜0.05),且均高于CON组表达,Akt在SAP大鼠回肠中表达逐渐减少,12h最少(P＜0.05),且均低于于CON组表达.结论 逆行注射5％牛磺胆酸钠12h后,可能是研究SAP大鼠肠道损伤最合适的时间点.%Objective The aims of this study were to clarify the of intestinal impairment in 5％ sodium taurocholate-induced severe acute pancreatitis (SAP) models and investigate a suitable time point for further study.Methods 59 male SD rats were randomly divided into 5 groups:the normal control group (n =12) receiving no treatment,the sham-operation group (CON group,n ＝12),SAP group (n =36,3,6,12 h groups,n =12).SAP model was induced by retrograde injection of 5％ sodium taurocholate into the biliopancreatic duct.Lipase,ascites and serum levels of amylase were measured in 3,6,12 hour.After sacrifice of experiment rats,pancreatic and ileum tissues were taken for pathological examination under light microscope,phosphorylated protein kinase B (p-Akt) and protein kinase B (Akt) expressions in ileum cell was studied by useing immunohistochemisty assay.Results After modelling,lipase,ascites,serum level of amylase,pathology score of ileum and pancreatic increased gradually,furthermore,the expressions of p-Akt in SAP group inreased gradually in SAP group,which were higher than those in CON group (P ＜ 0.01).The expressions of Akt in SAP group declined gradually in SAP group,which were lower than those in CON group (P ＜0.01).They achieved the peak in 12 hours after operation.Conclusion Based on the success rate of the mode,we conclude that 12 h after a retrograde injection of 5 ％ sodium taurocholate may be the most appropriate time point to study intestinal injury in SAP rats.