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重症急性胰腺炎并发肠道免疫功能障碍的实验研究
作者:周淼  兑丹华  敖宇  代静静 
单位:遵义医学院附属医院 普通外科, 贵州 遵义 563000
关键词:重症急性胰腺炎 肠道免疫屏障 高迁移率蛋白1 淋巴细胞 大鼠 
分类号:R576
出版年·卷·期(页码):2017·36·第四期(559-563)
摘要:

目的:探讨重症急性胰腺炎大鼠肠道免疫损伤的状况。方法:将48只SD大鼠随机分成假手术组(A组)和重症急性胰腺炎(severe acute pancreatitis,SAP)组(B组),各24只。A组开腹后仅翻动肠管;B组采用胰胆管逆行注射5%硫磺胆酸钠[0.1 ml·(100 g-1)]建立SAP模型。造模后6、12、24 h每组取8只大鼠,开腹取胰腺组织、回肠做病理学切片,光镜观察胰腺病理变化,光镜及电镜观察肠黏膜病理变化。酶联免疫吸附试验(ELISA)检测血清IL-1、IL-10的浓度,收集回肠Peyer's结,流式细胞术行CD3+、CD4+、CD8+细胞凋亡检测,RT-PCR检测回肠HMGB1mRNA含量。结果:B组各时间点胰腺病理学评分较A组升高(均P<0.01),B组随时间延长胰腺病理学评分逐渐升高(均P<0.05)。B组各时间点回肠HMGB1mRNA表达量均较假手术组升高(P<0.01或P<0.05),各时间点CD3+、CD4+、CD8+细胞的凋亡率较假手术组上升(P<0.01或P<0.05),回肠病理损伤较假手术组加重。结论:SAP大鼠并发肠道免疫功能障碍。

Objective:Objective:To investigate the status of intestinal immune damage in rats with severe acute pancreatitis. Methods:48 SD rats were randomly divided into sham operation group (A), SAP group (B), 24 rats in each. group A received laparotomy only without injection; group B retrograde pancreatic duct injection of 5% sodium taurocholate[0.1 ml·(100 g)-1] to establish SAP model. After modeling, 6 h, 12 h, 24 h, pancreas tissue and ileum were taken for pathological section, then the pathological changes were observed under light and electron microscopy to observe the pathological changes of pancreas and intestinal mucosa. Enzyme linked immunosorbent assay (ELISA) was performed to detect the concentration of serum IL-1, IL-10, the ileum s' Peyer nodes were collected, flow cytometry was performed for CD3+, CD4+, CD8+ cells apoptosis detection, and finally RT-PCR was carried out to detect the HMGB1mRNA content of ileum. Results:The pancreatic pathological score in group B was higher than that in group A (P<0.01), and the pancreatic pathological score was higher in group 6-24 h than in group B (P<0.05). HMGB1mRNA expression in ileum mucosa in group B at each time point was higher than that in the sham operation group (P<0.01 or P<0.05), the apoptosis rate of CD3+, CD4+ and CD8+ cells at each time point increased compared with the sham operation group (P<0.01 or P<0.05), ileum pathological injury increased compared with the sham operation group. Conclusion:Rats with severe acute pancreatitis have disturbance of intestinal immune function.

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