摘要
①目的 研究PC1 2神经细胞在炎症因子的刺激下 ,一氧化氮合酶 (NOS)底物精氨酸的来源 ,探讨减轻一氧化氮 (NO)介导的炎症反应途径。②方法 用神经生长因子 (NGF)处理PC1 2细胞使之分化成神经细胞 ,用IFNγ/TNFα激活 ,利用Northern及Western方法检测诱导型一氧化氮合酶 (iNOS)、精氨酸代琥珀酸合成酶 (AS)、精氨酸代琥珀酸裂解酶 (AL)、正性氨基酸转移酶 1和 2 (CAT 1、CAT 2 )mRNA及其酶蛋白的表达。同时检测地塞米松或dibutyrylcAMP对上述各种酶表达的影响。 ③结果 iNOSmRNA、ASmRNA和其酶蛋白明显被诱导 ;ALmRNA和其酶蛋白少量被诱导 ,同时产生大量的NO ;CAT 1和CAT 2没有被诱导。地塞米松或dibutyrylcAMP抑制iNOS的诱导及NO的合成 ,两者共同作用时 ,其抑制作用更明显。④结论 在细胞因子活化的PC1 2神经细胞中iNOS被诱导 ,产生大量的NO ;此时其底物精氨酸主要来源于精氨酸的再生系———瓜氨酸
Objective To elucidate the sources of arginine, substrate of nitric oxide synthase (NOS), with the stimulation to the neural PC12 cells of inflammatory factors, and to explore the ways to reduce inflammations induced by nitrate oxide(NO). Methods PC12 cells were differentiated with nerve growth factor, and then activated with interferon-γ (IFNγ) and tumor necrosis factor-α (TNFα). Northern and Western blotting was used to detect the expression of the mRNAs and enzymes of inducible NOS (iNOS), argininosuccinate synthetase(AS), argininosuccinate lyase(AL), cationic amino acid transporter-1 and -2 (CAT-1 and CAT-2). Results iNOS and AS mRNAs and proteins were markedly induced; while AL mRNA and proteins were weakly induced, and large amount of NO produced . CAT-1 and CAT-2 were not induced. Dexamethasone or dibutyryl cAMP inhibited the iNOS induction and NO production. The inhibition was even stronger when the two acted together. Conclusion In the cytokine-stimulated PC12 cells, iNOS is introduced and a large amount of NO is produced, when arginine, substrate of nitric oxide synthase comes from citrulline-arginine recycling.
出处
《青岛大学医学院学报》
CAS
2004年第2期121-124,127,共5页
Acta Academiae Medicinae Qingdao Universitatis
