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研究氨甲蝶呤 (MTX)在小鼠巨噬细胞 (RAW2 6 4 7)对一氧化氮合酶 (NOS)表达的抑制作用。本试验应用内毒素 (LPS)、干扰素 (IFNr)、MTX、二氨基 羟基嘧啶 (DAHP) ,四氢生物喋呤 (BH4 )刺激 8h后 ,用Northern杂交检测NOSmRNA水平和GTP CH 1mRNA水平表达 ,并用Griessreaction方法检测NO2- NO3- 浓度 ,用Duch方法测定GTP CH 1活性值。我们发现 ,MTX在小鼠巨噬细胞 (RAW2 6 4 7)对NOSmRNA水平和GTP CH 1mRNA水平表达有轻度抑制作用 ,对NO2- NO3- 浓度和GTP CH 1活性抑制作用较弱。MTX和DAHP联合应用不仅对NOSmRNA水平和GTP CH 1mRNA水平表达抑制作用较强 ,而且对NO2- NO3 浓度和GTP CH 1活性抑制较大。BH4的加入可以减少MTX和DAHP对一氧化氮 (NO)生成的抑制作用。
To investigate the inhibitory effect of methotrexate (MTX) on nitric oxide synthase (NOS) expression in murine macrophages (RAW26477). In this study, NOS mRNA level and GTP-CH 1 mRNA level were detected by Northern blot after stimulation with LPS, MTT, DAHP and BH4 for 8 hours, The concentration of NO2- NO3- was detected by Griessreaction method and the activity of GTP-CH1 was measured by Duch method. We found that MTX had a slight inhibitory effect on the expression of NOS mRNA and GTP CH 1 mRNA in mouse macrophages (RAW26477), with a weak inhibitory effect on NO2- NO3- and GTP CH1 activity. The combination of MTX and DAHP not only strongly inhibited the expression of NOS mRNA and GTP CH 1 mRNA, but also inhibited the activity of NO 2 -NO 3 and GTP CH 1. The addition of BH4 can reduce the inhibitory effect of MTX and DAHP on nitric oxide (NO) production.