论文部分内容阅读
目的：探讨鸟苷酸环化酶（ｓＧＣ）抑制剂甲基蓝（ＭＢ）对阿片耐受和依赖的阻断作用。方法：体外培养ｉＮＯＳ稳定表达的神经细胞做为模型，竞争性蛋白结合法、放免法和３ＨＡｒｇ转化３ＨＣｉｔ法检测胞内ｃＡＭＰ，ｃＧＭＰ水平和ＮＯＳ活性，激光共聚焦扫描技术测定胞内Ｃａ２＋浓度。观察ＭＢ对δ阿片激动剂ＤＰＤＰＥ和吗啡预处理细胞４８ｈ及纳洛酮戒断，对胞内ｃＧＭＰ，ｃＡＭＰ含量、［Ｃａ２＋］ｉ和ＮＯＳ活性的影响。结果：ＭＢ可明显抑制阿片激动剂长时程所致ｃＧＭＰ含量增加（Ｐ＜００１），对ｃＡＭＰ含量、［Ｃａ２＋］ｉ和ＮＯＳ活性的变化无影响。结论：ＭＢ通过抑制ｓＧＣ活性，使ＮＯｃＧＭＰ通路下调，减缓阿片耐受和依赖的发生。
Objective: To investigate the blockade of opiate tolerance and dependence by methyl blue (MB) inhibitor of guanylate cyclase (sGC). Methods: The iNOS-stably expressed neural cells were cultured in vitro as a model, competitive protein-binding assay, radioimmunoassay and 3H-Arg conversion 3H Cite assay intracellular cAMP, cGMP levels and NOS activity, confocal laser scanning confocal microscope Ca2 + concentration. To observe the effects of MB on δ-opioid agonist DPDPE and morphine preconditioning cells 48h and naloxone withdrew on intracellular cGMP, cAMP content, [Ca2 +] i and NOS activity. Results: MB inhibited the increase of cGMP content induced by long-term opioid agonist (P <001), but had no effect on the changes of cAMP content, [Ca2 +] i and NOS activity. Conclusion: MB inhibits the activity of sGC, down-regulates the NO-cGMP pathway and slows down the occurrence of opioid tolerance and dependence.