Differential expression of nitric oxide synthases in bacterial meningitis: Role of the inducible isoform for blood-brain barrier breakdown
Differential expression of nitric oxide synthases in bacterial meningitis: Role of the inducible isoform for blood-brain barrier breakdown
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DOI：
10.1086/320730
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发表时间：
2001-06-15
影响因子：
6.4
通讯作者：
Pfister, HW
中科院分区：
文献类型：
--
作者：
Winkler, F;Koedel, U;Pfister, HW
本研究的目的是确定一氧化氮（NO）合酶（NOS）异构体的差异表达和诱导型NOS（iNOS）在实验性肺炎球菌脑膜炎的病理生理相关性。通过使用逆转录-聚合酶链反应分析，免疫组织化学和蛋白质印迹，增加脑mRNA和增加的内皮型一氧化氮合酶（eNOS）和诱导型一氧化氮合酶的蛋白水平检测脑池内肺炎球菌接种后24小时。与野生型小鼠相比，iNOS缺陷型小鼠的血脑屏障（BBB）破坏显著减少。iNOS缺乏的这种有益作用与硝基酪氨酸免疫反应性的缺乏有关。此外，在缺乏iNOS的感染动物中，白细胞介素（IL）-1 β IL-6和肿瘤坏死因子-α的脑蛋白水平以及巨噬细胞炎性蛋白（MIP）-1 α和MIP-2的脑mRNA水平显著降低。这些结果表明：（1）在实验性细菌性脑膜炎的急性期，不仅iNOS而且eNOS都上调;（2）iNOS衍生的NO有助于该疾病中过氧亚硝酸盐的形成和BBB破坏。
The aim of the study was to determine the differential expression of nitric oxide (NO) synthase (NOS) isoforms and the pathophysiologic relevance of inducible NOS (iNOS) in experimental pneumococcal meningitis. By use of reverse transcription-polymerase chain reaction analysis, immunohistochemistry, and Western blotting, increased brain mRNA and increased protein levels of endothelial NOS (eNOS) and iNOS were detected 24 h after intracisternal pneumococcal inoculation. In iNOS-deficient mice, disruption of the blood-brain barrier (BBB) was significantly reduced, compared with that in wild-type mice. This beneficial effect of iNOS deficiency was associated with a lack of nitrotyrosine immunoreactivity. Furthermore, brain protein levels of interleukin (IL)-1 beta IL-6, and tumor necrosis factor-alpha and brain mRNA levels of macrophage inflammatory protein (MIP)-1 alpha and MIP-2 were significantly reduced in infected animals lacking iNOS. These findings suggest that (1) not only iNOS but also eNOS is up-regulated in the acute phase of experimental bacterial meningitis, and (2) iNOS-derived NO contributes to peroxynitrite formation and BBB breaching in this disease.