Division of Molecular Pharmacology, Department of Pharmacology, Jichi Medical University, Tochigi 329-0498, Japan.
Biochem Biophys Res Commun. 2010 Feb 19;392(4):603-7. doi: 10.1016/j.bbrc.2010.01.089. Epub 2010 Jan 25.
Cellular adaptations to chronic opioid treatment result in enhanced responsiveness of adenylate cyclase and an increase in forskolin- or agonist-stimulated cAMP production. It is, however, not known whether chaperone molecules such as heat shock proteins contribute to this adenylate cyclase sensitization. Here, we report that treatment of cells with geldanamycin, an inhibitor of heat shock protein 90 (Hsp90), led to effective attenuation of morphine-induced adenylate cyclase sensitization. In SK-N-SH human neuroblastoma cells, morphine significantly increased RNA transcript and protein levels of type I adenylate cyclase, leading to sensitization. Whole-genome tiling array analysis revealed that cAMP response element-binding protein, an important mediator for cellular adaptation to morphine, associated with the proximal promoter of Hsp90AB1 not only in SK-N-SH cells but also in rat PC12 and human embryonic kidney cells. Hsp90AB1 transcript and protein levels increased significantly during morphine treatment, and co-application of geldanamycin (0.1-10 nM) effectively suppressed the increase in forskolin-activated adenylate cyclase activation by 56%. Type I adenylate cyclase, but not Hsp90AB1, underwent significant degradation during geldanamycin treatment. These results indicate that Hsp90 is a new pharmacological target for the suppression of adenylate cyclase sensitization induced by chronic morphine treatment.
细胞对慢性阿片类药物治疗的适应导致腺苷酸环化酶的反应性增强，以及forskolin 或激动剂刺激的 cAMP 产生增加。然而，尚不清楚伴侣分子（如热休克蛋白）是否有助于这种腺苷酸环化酶敏化。在这里，我们报告细胞用格尔德霉素（热休克蛋白 90（Hsp90）的抑制剂）处理会导致吗啡诱导的腺苷酸环化酶敏化的有效衰减。在 SK-N-SH 人神经母细胞瘤细胞中，吗啡显着增加了 I 型腺苷酸环化酶的 RNA 转录物和蛋白水平，导致敏化。全基因组平铺阵列分析显示，cAMP 反应元件结合蛋白（一种对于细胞适应吗啡的重要介质）不仅在 SK-N-SH 细胞中，而且在大鼠 PC12 和人胚肾细胞中与 Hsp90AB1 的近端启动子相关。Hsp90AB1 的转录物和蛋白水平在吗啡处理期间显着增加，并且格尔德霉素（0.1-10 nM）的共同应用有效抑制了 forskolin 激活的腺苷酸环化酶激活增加了 56％。I 型腺苷酸环化酶，而不是 Hsp90AB1，在格尔德霉素处理期间经历了显着的降解。这些结果表明 Hsp90 是抑制慢性吗啡处理诱导的腺苷酸环化酶敏化的新的药理学靶标。
