首页> 外文期刊>American Journal of Physiology >Microtubule-active agents modify nitric oxide production in pulmonary artery endothelial cells.
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Microtubule-active agents modify nitric oxide production in pulmonary artery endothelial cells.

机译:微管活性剂改变肺动脉内皮细胞中一氧化氮的产生。

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The effects of specific microtubule-active agents on nitric oxide (NO) production were examined in pulmonary artery endothelial cells (PAEC). PAEC were incubated with taxol, which stabilizes microtubules, or nocodazole, which disrupts microtubules, or both for 2-4 h. We then examined NO production, endothelial NO synthase (eNOS) activity, and eNOS association with heat shock protein (HSP) 90. Incubation of PAEC with taxol (15 microM) for 2-4 h resulted in an increase in NO production, eNOS activity, and the amount of HSP90 binding to eNOS. Incubation of PAEC with nocodazole (50 microM) for 2-4 h induced a decrease in NO production, eNOS activity, and the amount of HSP90 binding to eNOS. The presence of taxol in the culture medium prevented the effects of nocodazole on NO production and eNOS activity in PAEC. Geldanamycin, a HSP90 inhibitor, prevented the taxol-induced increase in eNOS activity. Taxol and nocodazole did not affect eNOS, HSP90, and tubulin protein contents in PAEC, as detected using Western blot analysis. These results indicate that the polymerization state of the microtubule cytoskeleton regulates NO production and eNOS activity in PAEC. The changes in eNOS activity induced by modification of microtubules are due, at least in part, to the altered binding of HSP90 to eNOS protein.
机译:在肺动脉内皮细胞(PAEC)中检查了特定的微管活性剂对一氧化氮(NO)产生的影响。 PAEC与可稳定微管的紫杉醇或破坏微管的诺考达唑或两者孵育2-4小时。然后,我们检查了NO产生,内皮NO合酶(eNOS)活性以及eNOS与热休克蛋白(HSP)90的关联。PAEC与紫杉醇(15 microM)孵育2-4小时导致NO产生,eNOS活性增加,以及HSP90与eNOS结合的数量。 PAEC与诺考达唑(50 microM)孵育2-4小时可导致NO产量,eNOS活性以及HSP90与eNOS结合的数量减少。培养基中紫杉醇的存在阻止了诺考达唑对PAEC中NO产生和eNOS活性的影响。 HSP90抑制剂格尔德霉素可阻止紫杉醇诱导的eNOS活性增加。使用蛋白质印迹分析检测到紫杉醇和诺考达唑不会影响PAEC中eNOS,HSP90和微管蛋白的含量。这些结果表明,微管细胞骨架的聚合状态调节PAEC中的NO产生和eNOS活性。微管修饰引起的eNOS活性变化至少部分是由于HSP90与eNOS蛋白的结合改变。

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