Tung Nguyen Huu, Sun Kai, Fan Jing-Yu, Shoyama Yukihiro, Han Jing-Yan
Faculty of Pharmaceutical Sciences, Nagasaki International University, Nagasaki, Japan.
Microcirculation. 2014 Nov;21(8):688-95. doi: 10.1111/micc.12147.
NADPH oxidase activation results in ROS overproduction that is the pathological basis of I/R injury. This study aimed to investigate potential effects of ORG on I/R-induced ROS production in rat mesenteric microvasculature and underlying mechanisms.
Mesenteric I/R in Male Wistar rats (200~250 g) was induced by ligation of the mesenteric artery and vein for 10 minutes followed by reperfusion for 60 minutes by releasing of the occlusion. The rats were infused intravenously with or without ORG (5 mg/kg per hour) 10 minutes before ischemia (pretreatment) or 20 minutes after reperfusion (posttreatment). The DHR fluorescence intensity on, the leukocytes adherent to, and mast cell degranulation out of mesenteric venules were determined using an intravital microscope. NADPH oxidase subunit p47(phox) membrane translocation in intestine tissues was detected by Western blotting.
Pre- or posttreatment with ORG inhibited I/R-induced DHR fluorescence intensity on the venular walls and leukocytes adhesion, ORG pretreatment inhibited mast cell degranulation as well. Furthermore, the translocation of p47(phox) from cytosol to membrane was suppressed markedly by ORG after I/R.
The results suggested that ORG restrained I/R-induced ROS production, which might be correlated with its inhibitive effect on NADPH activation.
NADPH氧化酶激活导致活性氧(ROS)过量产生,这是缺血/再灌注(I/R)损伤的病理基础。本研究旨在探讨ORG对大鼠肠系膜微血管I/R诱导的ROS产生的潜在影响及其潜在机制。
通过结扎雄性Wistar大鼠(200~250 g)的肠系膜动静脉10分钟,然后松开结扎进行60分钟再灌注,诱导肠系膜I/R。在缺血前10分钟(预处理)或再灌注后20分钟(后处理),给大鼠静脉输注或不输注ORG(每小时5 mg/kg)。使用活体显微镜测定肠系膜小静脉上的DHR荧光强度、粘附的白细胞以及肥大细胞脱颗粒情况。通过蛋白质免疫印迹法检测肠组织中NADPH氧化酶亚基p47(phox)的膜转位。
ORG预处理或后处理均可抑制I/R诱导的小静脉壁上的DHR荧光强度和白细胞粘附,ORG预处理还可抑制肥大细胞脱颗粒。此外,I/R后ORG可显著抑制p47(phox)从胞质溶胶向膜的转位。
结果表明ORG可抑制I/R诱导的ROS产生,这可能与其对NADPH激活的抑制作用有关。