Increased activity and expression of iNOS in human duodenal enterocytes from patients with celiac disease
Increased activity and expression of iNOS in human duodenal enterocytes from patients with celiac disease
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DOI：
10.1152/ajpgi.00324.2001
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发表时间：
2002-08-01
影响因子：
4.5
通讯作者：
Long, RG
中科院分区：
文献类型：
--
作者：
Murray, IA;Daniels, I;Long, RG
本研究测定了从人十二指肠活检组织中分离的肠上皮细胞中一氧化氮合酶（NOS）的活性，以确定其在乳糜泻中的作用。患者分为肠易激综合征、缺铁性贫血、B-12/叶酸缺乏、治疗和未治疗的乳糜泻组。从所有组中分离的肠细胞显示出1400 W-可激发的Ca 2+非依赖性NOS活性，其最适pH水平和温度分别为9.4和37 ℃。蛋白质印迹法表明，肠上皮细胞表达的诱导型NOS蛋白和蛋白质与硝化酪氨酸残基，后者是一氧化氮驱动的过氧亚硝酸盐和/或自由基损伤的指示。内皮NOS仅见于固有层。乳糜泻患者的NOS活性高于其他患者组。对这种情况的治疗导致了活动的下降。酶联免疫吸附试验表明cGMP生产的肠细胞组分，但cGMP水平与NOS活性不相关。这些结果表明，诱导型一氧化氮合酶是组成型表达的人十二指肠上皮细胞，增加未经治疗的乳糜泻患者，并部分纠正时，这些患者进行治疗。我们没有发现任何证据支持一氧化氮在小肠内cGMP形成中的作用。此外，我们无法证明过氧亚硝酸盐/自由基损伤在乳糜泻的病理生理学中的作用。
The activity of nitric oxide synthase (NOS) was assayed in enterocytes isolated from human duodenal biopsies to determine its role in celiac disease. Patients were categorized into groups with irritable bowel syndrome, iron-deficiency anemia, B-12/folate deficiency, and treated and untreated celiac disease. Enterocytes isolated from all groups showed 1400W-inhibitable Ca2+-independent NOS activity with a pH level and temperature optimum of 9.4 and 37degreesC, respectively. Western blotting showed that enterocytes expressed the inducible NOS protein and proteins with nitrated tyrosine residues, the latter being indicative of nitric oxide-driven peroxynitrite and/or free-radical damage. Endothelial NOS was seen only in the lamina propria. Patients with celiac disease had higher NOS activity than other patient groups. Treatment of the condition led to a fall in activity. Enzyme-linked immunosorbent assay demonstrated cGMP production by the enterocyte fraction, but cGMP levels did not correlate with NOS activity. These results suggest that inducible NOS is constitutively expressed in human duodenal enterocytes, is increased in patients with untreated celiac disease, and is partially corrected when such patients are treated. We found no evidence to support a role for nitric oxide in the formation of cGMP within the small intestine. Furthermore, we were unable to demonstrate a role for peroxynitrite/free radical damage in the pathophysiology of celiac disease.