Department of Anesthesiology, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.
J Surg Res. 2010 Nov;164(1):e173-80. doi: 10.1016/j.jss.2009.10.003. Epub 2009 Oct 24.
Intestinal ischemia/reperfusion (I/R) injury is a critical condition associated with high morbidity and mortality. Our previous study showed that ischemic postconditioning (IPo) protects the intestinal mucosa from I/R injury. However, the precise molecular mechanisms of this event remain poorly elucidated. The aim of this study was to investigate the differentially expressed proteins of intestinal mucosa after intestinal I/R with or without IPo, and to explore the potential mechanisms of intestinal I/R injury and the protective effect of IPo in relation to the differential proteins.
Intestinal I/R injury was established by occluding the superior mesenteric artery (SMA) for 60 min followed by 60 min reperfusion. The rats were randomly allocated into one of three groups based upon the intervention (n = 8); sham : sham surgical preparation including isolation of the SMA without occlusion was performed; injury: there was no intervention either before or after SMA occlusion; IPo: three cycles of 30 s reperfusion-30 s reocclusion were imposed immediately upon reperfusion. A comparative proteomics approach with two-dimensional gel electrophoresis was used to isolate proteins in intestinal mucosa, the expression of which were regulated by I/R injury post-treated with or without IPo. The differentially displayed proteins were identified through matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS).
Image analysis revealed that an average of 1300 protein spots were detected on each gel; 16 and 9 proteins showing more than 1.5-fold difference were identified between the Sham versus Injury group and injury group versus IPo group, respectively. The identified proteins were functionally involved in the cellular processes of energy metabolism, anti-oxidation, and anti-apoptosis.
This study provided new clues for understanding the mechanisms of IPo against intestinal I/R injury.
肠缺血/再灌注(I/R)损伤是一种与高发病率和死亡率相关的危急情况。我们之前的研究表明,缺血后处理(IPo)可保护肠黏膜免受 I/R 损伤。然而,这一事件的确切分子机制仍未得到充分阐明。本研究旨在探讨肠 I/R 后有无 IPo 时肠黏膜差异表达蛋白,并探讨肠 I/R 损伤的潜在机制以及 IPo 与差异蛋白相关的保护作用。
通过阻断肠系膜上动脉(SMA)60 分钟后再灌注 60 分钟来建立肠 I/R 损伤。根据干预方式(n = 8),将大鼠随机分为三组:假手术组:仅进行 SMA 分离而不阻断;损伤组:在 SMA 阻断前后均无干预;IPo 组:再灌注后立即进行 3 个 30 秒再灌注-30 秒再闭塞的循环。采用二维凝胶电泳的比较蛋白质组学方法分离肠黏膜中的蛋白质,这些蛋白质的表达受到 I/R 损伤后有无 IPo 处理的调节。通过基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF-MS)鉴定差异表达蛋白。
图像分析显示,每张凝胶上平均可检测到 1300 个蛋白点;在 Sham 与 Injury 组之间和 Injury 与 IPo 组之间,分别有 16 个和 9 个蛋白的表达差异超过 1.5 倍。鉴定的蛋白功能涉及细胞能量代谢、抗氧化和抗凋亡过程。
本研究为理解 IPo 对抗肠 I/R 损伤的机制提供了新线索。