HSP90 inhibition by 17-DMAG reduces inflammation in J774 macrophages through suppression of Akt and nuclear factor-κB pathways
HSP90 inhibition by 17-DMAG reduces inflammation in J774 macrophages through suppression of Akt and nuclear factor-κB pathways
复制标题
DOI：
10.1007/s00011-012-0442-x
复制
发表时间：
2012-05-01
影响因子：
6.7
通讯作者：
Rylander, M. Nichole
中科院分区：
文献类型：
--
作者：
Shimp, Samuel K., III;Parson, Carl D.;Rylander, M. Nichole
本研究旨在确定热休克蛋白90（HSP 90）抑制剂是否通过减少免疫刺激的巨噬细胞中的核因子（NF）-κ B和Akt信号通路来减少促炎介质的产生（0.01、0.1或1 μ M）。通过蛋白质印迹法测量全细胞裂解物中Akt、κ B激酶（IKK）抑制剂和热休克蛋白的表达。通过ELISA在全细胞裂解物中测量磷酸化Akt和κ B抑制剂（I κ B）。分析细胞上清液中的白细胞介素（IL）-6，肿瘤坏死因子（TNF）-α和一氧化氮（NO）。免疫荧光检测NF-κ B和热休克因子（HSF）-1的易位。17-DMAG处理细胞可降低免疫刺激细胞中Akt和IKK的表达。17-DMAG可减少NF-κ B的核转位，减少免疫刺激产生的IL-6、TNF-α和NO，但不减少诱导型一氧化氮合酶的表达。我们的研究表明，免疫介导的NF-κ B炎症级联反应可被HSP 90抑制剂17-DMAG阻断。由于HSP 90与许多促炎激酶级联反应的广泛相互作用，抑制HSP 90可能提供一种减轻慢性炎症的新方法。
This study was designed to determine whether inhibition of heat shock protein 90 (HSP90) reduces pro-inflammatory mediator production by decreasing the nuclear factor (NF)-kappa B and Akt signaling pathways in immune-stimulated macrophages.J774A.1 murine macrophages were treated with the HSP90 inhibitor 17-DMAG (0.01, 0.1 or 1 mu M) prior to immune stimulation with lipopolysaccharide and interferon-gamma. Expression of Akt, inhibitor of kappa B kinase (IKK), and heat shock proteins were measured in whole cell lysates by Western blotting. Phosphorylated Akt and inhibitor of kappa B (I kappa B) were measured in whole cell lysates by ELISA. Cell supernatants were analyzed for interleukin (IL)-6, tumor necrosis factor (TNF)-alpha and nitric oxide (NO). Translocation of NF-kappa B and heat shock factor (HSF)-1 was assessed by immunofluorescence.Treating cells with 17-DMAG reduced expression of Akt and IKK in immune-stimulated cells. 17-DMAG reduced nuclear translocation of NF-kappa B and reduced immune-stimulated production of IL-6, TNF-alpha and NO, but did not decrease inducible nitric oxide synthase expression.Our studies show that the immune-mediated NF-kappa B inflammatory cascade is blocked by the HSP90 inhibitor 17-DMAG. Due to the broad interaction of HSP90 with many pro-inflammatory kinase cascades, inhibition of HSP90 may provide a novel approach to reducing chronic inflammation.