含益生元肠内营养对重症急性胰腺炎大鼠肠紧密结合蛋白occludin的作用

Effect of enteral nutrition with prebiotics on integral membrane protein occludin in intestine in severe acute pancreatitis rats

目的探讨早期给予含益生元（低聚半乳糖）肠内营养（EN）对重症急性胰腺炎（SAP）大鼠肠紧密结合蛋白oecludin蛋白表达以及mRNA表达的作用。方法将64只SD大鼠随机分成8组（n=8）：假手术对照4d和7d组、SAP造模＋肠内营养（SAP＋EN）4d和7d组、SAP造模＋含益生元肠内营养（SAP＋PRE-EN）4d和7d组、SAP造模＋肠外营养（SAP4-PN）4d和7d组。胰腺被膜下均匀注射3．8％牛磺胆酸钠1ml建立SAP大鼠模型，采用免疫组织化学法检测occludin蛋白表达，实时荧光定量PCR法检测occludinmRNA含量。结果各SAP组oecludin蛋白表达和mRNA含量均显著低于假手术对照组（P〈0．01），SAP＋EN组和SAP＋PRE-EN组显著高于SAP＋PN组（P〈0．05），SAP＋PRE．EN组显著高于SAP＋EN组（P〈0．05）。Occludin蛋白表达和mRNA含量呈显著正相关（r=0．83，P〈0．01）。结论含益生元肠内营养可以增强肠黏膜紧密结合蛋白occludin的表达，其作用与上调occludinmRNA水平有关。

Objective To explore the effect of enteral nutrition （EN） with prebiotics on integral membrane protein occludin in severe acute pancreatitis （SAP） rats. Methods SAP-specific EN formula was prepared based on the nutritional and metabolic characteristics of SAP, and prebioitcs galactooligosaccharides （GOS） was used. Sixty- four SD rats were divided randomly into 8 groups （n = 8） ： sham operation control 4 days and 7 days groups, SAP with EN （ SAP ＋ EN） 4 days and 7 days groups, SAP with prebiotics-supplemented EN （SAP ＋ PRE-EN） 4 days and 7 days groups, and SAP with parenteral nutrition （ SAP ＋ PN） 4 days and 7 days groups. The SAP model in rats was established by injection of I ml of 3.8% sodium taurocholate beneath the pancreatic capsule. The nutritional therapy began after SAP model was established. The rats were killed on Day 4 and Day 7 respectively. The expression of oc- cludin protein in intestinal mucosa was analyzed by immunohistochemistry method, and occludin mRNA level was de- tected by real-time fluorescent quantitative PCR technique. Results At each time point, occludin protein and mR- NA levels in all SAP model groups were significantly higher than in sham operation control group （ P 〈 0. 01 ）, the levels in SAP ＋ EN and SAP ＋ PRE-EN groups were significantly higher than SAP ＋ PN groups （ P 〈 0.05 ）, and the levels in SAP ＋ PRE-EN groups increased significantly compared with SAP ＋ EN groups （ P 〈 0. 05 ）. A signifi- cantly positive correlation was observed between occludin protein density and occludin mRNA level （r = 0. 83, P 〈 0. 01 ）. Conclusion EN supplemented with prebiotics has the effect of maintaining integrity of epithelial cells by increasing occludin protein expression, and the effect is related with the up-regulation of occludin mRNA.