Takahama Umeo, Hirota Sachiko, Takayuki Oniki
Department of Bioscience, Kyushu Dental College, Kitakyushu, Japan.
Methods Enzymol. 2008;440:381-96. doi: 10.1016/S0076-6879(07)00824-5.
Nitrate is secreted into the human oral cavity as a salivary component. The nitrate is transformed to nitrite and nitric oxide (NO) by oral bacteria. NO is oxidized by O(2) producing NO(2) and N(2)O(3) and also by O(2)(-) producing ONOO(-). Salivary peroxidase can oxidize nitrite and NO to NO(2) or its equivalent in the oral cavity. Nitrite dissolved in saliva is mixed with gastric juice, generating nitrous acid that is transformed to NO and NO(2) via N(2)O(3) by self-decomposition. In addition, nitrous acid can react with ascorbic acid and phenolics producing NO and with H(2)O(2) producing ONOOH. This chapter deals with the detection of reactive nitrogen oxide species (RNOS), especially NO, N(2)O(3), NO(2), and ONOO(-)/ONOOH, in mixed whole saliva and acidified saliva using fluorescent probes and spin-trapping reagents. It is also shown that measurements of nitration and oxygen consumption are useful in studying the formation and scavenging of RNOS in the aforementioned systems.
硝酸盐作为唾液成分分泌到人体口腔中。硝酸盐被口腔细菌转化为亚硝酸盐和一氧化氮(NO)。NO被O₂氧化生成NO₂和N₂O₃,也被超氧阴离子(O₂⁻)氧化生成过氧亚硝酸根(ONOO⁻)。唾液过氧化物酶可在口腔中将亚硝酸盐和NO氧化为NO₂或其等效物。溶解在唾液中的亚硝酸盐与胃液混合,生成亚硝酸,亚硝酸通过自身分解经由N₂O₃转化为NO和NO₂。此外,亚硝酸可与抗坏血酸和酚类物质反应生成NO,并与过氧化氢(H₂O₂)反应生成过氧亚硝酸(ONOOH)。本章介绍了使用荧光探针和自旋捕获试剂检测混合全唾液和酸化唾液中活性氮氧化物(RNOS),特别是NO、N₂O₃、NO₂和ONOO⁻/ONOOH的方法。还表明,硝化作用和耗氧量的测量对于研究上述系统中RNOS的形成和清除是有用的。