Mitchell J A, Kohlhaas K L, Matsumoto T, Pollock J S, Förstermann U, Warner T D, Schmidt H H, Murad F
Department 47S, Abbott Laboratories, Abbott Park, Illinois 60064.
Mol Pharmacol. 1992 Jun;41(6):1163-8.
Lipopolysaccharide (LPS), either alone or in combination with cytokines, induces nitric oxide (NO) synthase activity in cells that normally release little or no NO. In arterial smooth muscle cells and various macrophage cell lines, NO synthase activity is induced after several hours of incubation with LPS. In brain, NADPH-dependent diaphorase activity has been associated with constitutive NO synthase. Here we show that incubation of rat aorta or cultured macrophages with LPS causes a time-dependent induction of NO synthase. The NO synthase activity in both rat aorta and macrophages was calcium independent and inhibited by NG-monomethyl-L-arginine and NG-nitro-L-arginine. We also found that LPS caused a time-dependent induction in NADPH-dependent diaphorase activity in both rat aorta and cultured macrophages. The diaphorase activity was mainly NADPH dependent and NADH independent. NO synthase activity and NADPH-diaphorase activity in crude cytosol from LPS-treated macrophages were found to co-purify, using 2',5'-ADP-Sepharose followed by Superose-6 gel permeation chromatography.
脂多糖（LPS）单独或与细胞因子联合使用时，可在通常释放很少或不释放一氧化氮（NO）的细胞中诱导一氧化氮合酶活性。在动脉平滑肌细胞和各种巨噬细胞系中，与LPS孵育数小时后可诱导一氧化氮合酶活性。在大脑中，NADPH依赖性黄递酶活性与组成型一氧化氮合酶有关。在此我们表明，用LPS孵育大鼠主动脉或培养的巨噬细胞会导致一氧化氮合酶的时间依赖性诱导。大鼠主动脉和巨噬细胞中的一氧化氮合酶活性均不依赖于钙，并被NG-单甲基-L-精氨酸和NG-硝基-L-精氨酸抑制。我们还发现，LPS在大鼠主动脉和培养的巨噬细胞中均导致NADPH依赖性黄递酶活性的时间依赖性诱导。黄递酶活性主要依赖于NADPH而不依赖于NADH。使用2'，5'-ADP-琼脂糖随后进行Superose-6凝胶渗透色谱法，发现来自LPS处理的巨噬细胞的粗提物中的一氧化氮合酶活性和NADPH-黄递酶活性可共同纯化。
