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目的:探讨乌司他丁对脓毒症大鼠受损肠屏障的保护作用及其可能机制。方法:采用盲肠结扎穿孔术(CLP)制作脓毒症模型,36只雄性SD大鼠随机分为对照组、脓毒症组和乌司他丁组,每组各12只大鼠。乌司他丁组实验前15min大鼠尾静脉注射乌司他丁(20万U/kg),12h后重复给药,对照组、脓毒症组于上述时间点注射等量生理盐水,建模后6、12、24、48h大鼠尾静脉采血测定TNF-α、IL-6的质量浓度;留取肠组织行石蜡包埋,苏木精-伊红染色及Bax、Caspase-3凋亡蛋白检测;透射电镜下观察肠黏膜内部超微结构改变。结果:脓毒症组大鼠血清中TNF-α、IL-6质量浓度明显升高(与对照组比较,P<0.01;与乌司他丁组比较,P<0.05),而乌司他丁组较对照组高,但明显低于脓毒症组(P<0.05);苏木精-伊红染色可见脓毒症组肠黏膜水肿、充血,炎症细胞浸润,而乌司他丁组较脓毒症组明显减轻;凋亡蛋白Bax、Caspase-3表达较乌司他丁组明显减少(P<0.05);通过透射电镜观察脓毒症组肠上皮及毛细血管基底膜肿胀、细胞间紧密连接明显开放增宽,中性粒细胞侵润,肠黏膜细胞绒毛倒伏及脱落、线粒体肿胀及空化,而乌司他丁组明显减轻。结论:乌司他丁改善脓毒症大鼠肠黏膜高炎症状态,减少促炎因子的释放,降低凋亡蛋白Bax、Caspase-3的表达,减少肠黏膜上皮细胞凋亡,对肠屏障有保护作用。
Objective: To investigate the protective effect of ulinastatin on the damaged gut barrier in septic rats and its possible mechanism. Methods: Sepsis model was made by cecal ligation and puncture (CLP). Thirty-six male Sprague-Dawley rats were randomly divided into control group, sepsis group and ulinastatin group, with 12 rats in each group. Ulinastatin (200,000 U / kg) was injected into tail vein of rats in ulinastatin group 15 minutes before experiment and repeated 12 hours after injection. In control group and sepsis group, normal saline was injected into the sepsis group After 6, 12, 24, 48 hours, blood samples were collected from the tail vein to determine the concentration of TNF-α and IL-6. Paraffin-embedded, hematoxylin-eosin and Bax, Caspase- The ultrastructural changes of intestinal mucosa were observed under transmission electron microscope. Results: Serum levels of TNF-α and IL-6 in sepsis group were significantly increased (P <0.01 compared with control group, P <0.05 compared with ulinastatin group) Group was higher than the control group, but significantly lower than the sepsis group (P <0.05); hematoxylin-eosin staining showed intestinal mucosal edema, congestion, infiltration of inflammatory cells in sepsis group, and ulinastatin group pus The expression of Bax and Caspase-3 in the sepsis group was significantly lower than that in the ulinastatin group (P <0.05). The transmission electron microscope was used to observe the swollen intestinal epithelium and capillary basement membrane in the sepsis group and the tight intercellular junctions Obviously widened open, neutrophil infiltration, lumbar mucosal cells lint and fall off, mitochondria swelling and cavitation, and ulinastatin group was significantly reduced. Conclusion: Ulinastatin can improve the inflammatory state of intestinal mucosa in septic rats, reduce the release of proinflammatory cytokines, decrease the expression of Bax and Caspase-3, decrease the apoptosis of intestinal epithelial cells and protect the gut barrier effect.