Comparisons of Nitric Oxide Synthases in Normal Human Placenta from 37 to 41 Weeks Gestation: Qualitative and Quantitative Analyses.

Otsubo, Hori, Nishino, Araki
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Abstract

> Objective: The purpose of this study was to determine the type of nitric oxide synthase isoform and to measure the quantities of nitric oxide synthase mRNA in the human placenta. Methods: The isoforms of nitric oxide synthase were determined in ten placentas of normal pregnant women using information regarding calcium dependence and inhibition by arginine analogs and findings from Western blot analyses and reverse transcriptase-polymerase chain reaction. Results: The activity of nitric oxide synthase was largely calcium dependent, although a small element of calcium-independent activity was observed. A stronger inhibition was shown with Nomega-nitro-l-arginine than with Nomega-monomethyl-l-arginine. On Western blots endothelial nitric oxide synthase was detected as a band of 140 kilodaltons, without evidence of inducible nitric oxide synthase. Messenger RNA of endothelial nitric oxide synthase was readily detected by reverse transcriptase-polymerase chain reaction, but that of inducible nitric oxide synthase was barely detected. The quantity of mRNA of inducible nitric oxide synthase was about 100-fold less than that of endothelial nitric oxide synthase. Conclusions: These results point to the constitutive endothelial type as the predominant isoform of nitric oxide synthase in human placenta from 37 to 41 weeks gestation, although protein and mRNA of inducible nitric oxide synthase may exist.

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妊娠37 ~ 41周正常人胎盘一氧化氮合酶的比较:定性和定量分析。
目的:测定人胎盘中一氧化氮合酶异构体的类型和一氧化氮合酶mRNA的含量。方法:应用钙依赖、精氨酸类似物抑制、Western blot和逆转录聚合酶链反应等方法,对10例正常孕妇胎盘一氧化氮合酶的同工型进行测定。结果:一氧化氮合酶的活性在很大程度上依赖于钙,尽管观察到少量不依赖钙的活性。诺米加-硝基-l-精氨酸比诺米加-单甲基-l-精氨酸有更强的抑制作用。Western blots检测到内皮型一氧化氮合酶为140千道尔顿带,未发现诱导型一氧化氮合酶。逆转录-聚合酶链式反应可以检测到内皮型一氧化氮合酶的信使RNA,而诱导型一氧化氮合酶的信使RNA几乎无法检测到。诱导型一氧化氮合酶mRNA表达量约为内皮型一氧化氮合酶mRNA表达量的100倍。结论:这些结果表明,尽管诱导型一氧化氮合酶的蛋白和mRNA可能存在,但在妊娠37至41周的人胎盘中,一氧化氮合酶的主要亚型是构成型内皮型。
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