炎症因子对重症急性胰腺炎大鼠肠道屏障的影响

发布时间：2018-03-26 18:12

  本文选题：重症急性胰腺炎　切入点：肠道免疫屏障　出处：《皖南医学院》2016年硕士论文

【摘要】：目的:建立大鼠重症急性胰腺炎(SAP)模型,动态的观察SAP大鼠胰腺、肠组织病理、血清TNF-α、IL-6浓度变化,以及小肠组织匀浆TNF-α、IL-6和小肠粘液SIgA变化,进一步探讨SAP时炎症因子的变化以及肠道局部炎症因子与肠道粘膜屏障的关系。方法:48只健康成年雄性SD大鼠,体重250±30g,随机分成2组:假手术组(A组,N=24)、SAP组(B组,N=24)各组再按术后6、12、24h小时时段随机分为3个小组。每个时间点8只。实验前各组大鼠均禁食12h,可自由饮水。SAP组:以5%牛磺胆酸钠(0.1ml/100g)逆行胰胆管注射,诱导SAP模型;A组:开腹后以生理盐水(0.1ml/100g体重)逆行胰胆管注射,术后于大鼠大腿根部皮下注射0.9%氯化钠溶液2ml/kg,以补充丢失液体,大鼠仍然禁食,清醒后可自由饮水。各组24只大鼠分6h,12h,24h三个时间点,每个时间点8只开腹检验。肉眼观察胰腺和肠大体改变,腹主动脉采集血液标本5ml,离心后取血清,血清样本保存于-20℃,以备后续检测用,生化自动分析仪法检测血清淀粉酶,并采用ELISA法测定血清TNF-α、IL-6等细胞因子以及小肠组织匀浆TNF-α、IL-6、小肠粘液SIgA,制作胰腺和回肠组织标本,HE染色用于胰腺和回肠组织损伤的病理学评价,实验所得数据利用SPSS17.0统计软件进行统计学分析。结果:1、SAP组与假手术组比较各时间点(术后6h、12h、24h)血清AMY、IL-6、TNF-α水平显著升高(P0.01)。2、SAP组各时间点胰腺和回肠组织病理学评分较假手术组均升高(P0.01),且SAP组胰腺和回肠组织病理学评分随时间延长而增加。3、SAP组大鼠各时间点小肠粘液SIgA低于假手术组。4、SAP组与假手术组相比较,随造模时间的延长,小肠黏液SIgA浓度降低,小肠组织匀浆IL-6、TNF-α水平及小肠组织病理学评分升高(P0.01)。5、小肠组织匀浆TNF—α、IL-6浓度与小肠黏液SIgA之间具有明显的负相关(P0.01),与小肠组织病理学评分具有明显的正相关(P0.0 1)。结论:1、采用逆行性胰胆管注射5%牛磺胆酸钠可成功制备大鼠重症急性胰腺炎模型,此实验方法较稳定,重复性好。2、SAP时细胞因子TNF-α和IL-6参与了SAP的发生和发展,TNF-α和IL-6水平的高低可能作为胰腺炎预后的预测指标。3、SAP大鼠早期即可发生肠黏膜屏障功能障碍,免疫屏障受损,表现为SIgA下降。4、SAP造成胰腺损伤的同时破坏肠道粘膜免疫屏障,随着时间的延长损伤加重,其表现为肠粘液SIgA减少,肠道局部炎症因子TNF-α、IL-6增加。
[Abstract]:Objective: to establish a rat model of severe acute pancreatitis (SAP) and observe dynamically the changes of pancreatic and intestinal histopathology, serum TNF- 伪 IL-6 concentration, small intestinal homogenate TNF- 伪 IL-6 and intestinal mucus SIgA in SAP rats. To further investigate the changes of inflammatory factors during SAP and the relationship between intestinal inflammatory factors and intestinal mucosal barrier. Weight 250 卤30g, randomly divided into two groups: sham operation group A group A group A group SAP group B group (n = 24). Each group was randomly divided into three groups according to 612h 24 hours after operation, 8 rats at each time point. Before the experiment, rats in each group were fasting for 12 hours, free drinking water. SAP group: 5%. Sodium taurocholate 0.1 ml / 100 g retrograde cholangiopancreatic injection, Group A of induced SAP model: after operation, the rats were injected with 0.1 ml / 100g body weight of normal saline) retrograde cholangiopancreatopancreatic injection. After the operation, 0.9% sodium chloride solution was subcutaneously injected into the root of the thigh of rats to supplement the lost fluid, and the rats still fasted. 24 rats in each group were divided into 6 hours and 12 hours for 24 hours, 8 rats were examined at each time point. Gross changes of pancreas and intestine were observed with naked eyes. Blood samples were collected from abdominal aorta at 5 ml, and serum samples were collected after centrifugation. The serum samples were stored at -20 鈩，

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