Nitric oxide donor-induced hyperpermeability of cultured intestinal epithelial monolayers: role of superoxide radical, hydroxyl radical, and peroxynitrite.
Nitric oxide donor-induced hyperpermeability of cultured intestinal epithelial monolayers: role of superoxide radical, hydroxyl radical, and peroxynitrite.
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一氧化氮供体诱导的培养肠上皮单层的通透性过高：超氧自由基、羟自由基和过氧亚硝酸盐的作用。
DOI：
10.1016/s0304-4165(98)00072-5
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发表时间：
1998
期刊：
影响因子：
--
通讯作者：
Fink,MP
中科院分区：
文献类型：
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作者：
Menconi,MJ;Unno,N;Smith,M;Aguirre,DE;Fink,MP
一氧化氮 (NO·) 的许多细胞病变效应是由过氧亚硝酸盐 (PN) 介导的，过氧亚硝酸盐 (PN) 是 NO·与超氧自由基 (O·−2) 之间反应的产物。在本研究中，我们研究了 PN、O·−2 和羟基自由基 (OH·) 作为 NO·供体 S-亚硝基-N-乙酰青霉胺 (SNAP) 和 PN 产生剂 3-吗啉代亚胺 (SIN-1) 诱导的上皮通透性过高介质的作用。 Caco-2BBe肠细胞单层在两室室的可渗透支持物上生长。上皮渗透性（以荧光素二磺酸从顶端到基底外侧的通量测量）在与 5.0 mM SNAP 或 SIN-1 孵育 24 小时后增加。添加NO·清除剂2-(4-羧基苯基)-4,4,5,5-四甲基咪唑啉-1-氧基-3-氧化物，或O·−2清除剂Tiron，减少了两种供体化合物引起的渗透性增加。 SNAP 诱导的通透性增加被别嘌呤醇（一种黄嘌呤氧化酶抑制剂（内源性 O·−2 的来源））阻止。二乙基二硫代氨基甲酸酯（一种超氧化物歧化酶抑制剂）和邻苯三酚（一种 O·−2 生成剂）可增强 SNAP 诱导的通透性增加。添加 PN 清除剂去铁胺、尿酸盐或谷胱甘肽，或 OH·清除剂甘露醇，可减弱 SNAP 和 SIN-1 诱导的通透性增加。两种供体化合物均降低了细胞内谷胱甘肽和蛋白质结合的巯基的水平，表明产生了有效的氧化剂。这些结果支持 PN 以及可能的 OH· 在 NO·供体诱导的肠上皮通透性过高的发病机制中发挥作用。
Many of the cytopathic effects of nitric oxide (NO·) are mediated by peroxynitrite (PN), a product of the reaction between NO·and superoxide radical (O·−2). In the present study, we investigated the role of PN, O·−2and hydroxyl radical (OH·) as mediators of epithelial hyperpermeability induced by the NO·donor, S-nitroso-N-acetylpenicillamine (SNAP), and the PN generator, 3-morpholinosydnonimine (SIN-1). Caco-2BBeenterocytic monolayers were grown on permeable supports in bicameral chambers. Epithelial permeability, measured as the apical-to-basolateral flux of fluorescein disulfonic acid, increased after 24 h of incubation with 5.0 mM SNAP or SIN-1. Addition of 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide, an NO·scavenger, or Tiron, an O·−2scavenger, reduced the increase in permeability induced by both donor compounds. The SNAP-induced increase in permeability was prevented by allopurinol, an inhibitor of xanthine oxidase (a source of endogenous O·−2). Diethyldithiocarbamate, a superoxide dismutase inhibitor, and pyrogallol, an O·−2generator, potentiated the increase in permeability induced by SNAP. Addition of the PN scavengers deferoxamine, urate, or glutathione, or the OH·scavenger mannitol, attenuated the increase in permeability induced by both SNAP and SIN-1. Both donor compounds decreased intracellular levels of glutathione and protein-bound sulfhydryl groups, suggesting the generation of a potent oxidant. These results support a role for PN, and possibly OH·, in the pathogenesis of NO·donor-induced intestinal epithelial hyperpermeability.