Molecular Gastroenterology and Hepatology, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto 602-8566, Japan.
Mol Med Rep. 2011 Jan-Feb;4(1):81-6. doi: 10.3892/mmr.2010.386. Epub 2010 Oct 21.
Protease-activated receptors (PARs) are widely recognized for their modulatory properties during inflammation. The aim of the present study was to examine the role of PAR-2 on ischemia/reperfusion-induced small intestinal injury in rats. Intestinal damage was induced in male Wistar rats by clamping both the superior mesenteric artery and the celiac trunk for 30 min, followed by reperfusion for 60 min. Expression of PAR-2 in the intestinal mucosa was estimated by Western blot analysis and real-time PCR. Anti-rat PAR-2 cleavage site (PCS) antibody was intraperitoneally administered to the rats 1 h before the vascular clamping. The intestinal mucosal injury and inflammation were evaluated by biochemical markers and histological findings. Thiobarbituric acid (TBA) reactive substances and tissue-associated myeloperoxidase (MPO) activity were measured in the intestinal mucosa as indices of lipid peroxidation and neutrophil infiltration, respectively. Expression of cytokine-induced neutrophil chemoattractant-1 (CINC-1) in intestinal mucosa was measured by enzyme-linked immunosorbent assay. Expression of PAR-2 mRNA and protein in the intestinal mucosa was increased after reperfusion following ischemia. Reperfusion after ischemia resulted in an increase in luminal protein concentrations, hemoglobin concentrations, TBA reactive substances, MPO activity and CINC-1 protein. Pre-treatment with anti-rat PCS antibody significantly inhibited the increases in these parameters. These results suggest that PAR-2 plays an important role in the pathogenesis of ischemia/reperfusion-induced intestinal injury.
蛋白酶激活受体（PARs）在炎症过程中的调节作用已得到广泛认可。本研究旨在探讨 PAR-2 在大鼠肠缺血/再灌注损伤中的作用。雄性 Wistar 大鼠通过夹闭肠系膜上动脉和腹腔干 30 分钟，然后再灌注 60 分钟，诱导肠道损伤。通过 Western blot 分析和实时 PCR 评估肠道黏膜中 PAR-2 的表达。在血管夹闭前 1 小时，腹腔内给予抗大鼠 PAR-2 裂解位点（PCS）抗体。通过生化标志物和组织学发现评估肠道黏膜损伤和炎症。肠道黏膜中的硫代巴比妥酸（TBA）反应性物质和组织相关髓过氧化物酶（MPO）活性分别作为脂质过氧化和中性粒细胞浸润的指标进行测量。通过酶联免疫吸附试验测量肠道黏膜中细胞因子诱导的中性粒细胞趋化因子-1（CINC-1）的表达。缺血后再灌注导致肠道黏膜中 PAR-2 mRNA 和蛋白的表达增加。缺血后再灌注导致腔蛋白浓度、血红蛋白浓度、TBA 反应性物质、MPO 活性和 CINC-1 蛋白增加。抗大鼠 PCS 抗体预处理可显著抑制这些参数的增加。这些结果表明 PAR-2 在缺血/再灌注诱导的肠道损伤发病机制中起重要作用。
