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目的:通过氨基胍(AG)干预内毒素活化小鼠巨噬细胞RAW264.7模型,观察NO的产生、左旋精氨酸(L-Arg)转运以及碱性氨基酸转运体-2(CAT-2)mRNA的表达,旨在研究氨基胍对L-Arg跨膜转运及CAT-2表达的影响。方法:实验分为四组,即对照组,LPS对照组,AG组,AG+LPS组。接种RAW264.7细胞于培养板后,37℃、5%CO2培养箱培养24 h,将AG组和AG+LPS组换以含AG 1 mmol/L的DMEM培养液,30 min后四组均换以新鲜DMEM培养液,其中LPS对照组和AG+LPS组加入LPS,继续培养18 h,检测细胞合成NO水平、L-Arg摄取率和CAT-2 mRNA表达。结果:与对照组比较,AG预处理后的RAW264.7细胞,经LPS活化后NO水平、L-Arg摄取率、CAT-2 mRNA水平显著降低。结论:AG作为一种特异的诱生型一氧化氮合酶(iNOS)抑制剂,不仅可抑制iNOS的活性,而且还可以从基因水平上抑制L-Arg转运体CAT-2的表达,进而阻断L-Arg的跨膜转运和NO合成。
OBJECTIVE: To investigate the effect of endotoxin on the macrophage RAW264.7 model induced by nitric oxide (AG), and to observe the effects of NO production, L-arginine (L-Arg) transport, and basic amino acid transporter- The aim of this study is to investigate the effect of aminoguanidine on the transmembrane transport of L-Arg and the expression of CAT-2. Methods: The experiment was divided into four groups: control group, LPS control group, AG group and AG + LPS group. Inoculation of RAW264.7 cells in culture plate, 37 ℃, 5% CO2 incubator for 24 h, the AG group and AG + LPS group with AG 1 mmol / L DMEM medium, 30 min after the four groups were replaced LPS and LPS were added into fresh DMEM culture medium, LPS control group and AG + LPS group were added into LPS for 18 h, and NO synthesis, L-Arg uptake and CAT-2 mRNA expression were detected. Results: Compared with the control group, RAW264.7 cells pretreated with AG significantly decreased NO, L-Arg uptake and CAT-2 mRNA levels after LPS activation. Conclusion: As a specific inhibitor of inducible nitric oxide synthase (iNOS), AG can not only inhibit the activity of iNOS, but also inhibit the expression of L-Arg transporter CAT-2 genetically Transmembrane transport and NO synthesis of L-Arg.