Lipopolysaccharide‐induced histone deacetylase activity increases paracellular permeability in human lung microvascular endothelial cells (695.4)
Lipopolysaccharide‐induced histone deacetylase activity increases paracellular permeability in human lung microvascular endothelial cells (695.4)
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DOI：
10.1096/fasebj.28.1_supplement.695.4
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发表时间：
2014-04
期刊：
影响因子：
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通讯作者：
Atul D Joshi;N. Barabutis;G. Thangjam;Charalampos Birmpas;J. Catravas
中科院分区：
文献类型：
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作者：
Atul D Joshi;N. Barabutis;G. Thangjam;Charalampos Birmpas;J. Catravas
目的：脂多糖导致内皮屏障破坏并增加跨内皮渗透性。抑制热休克蛋白90（Hsp 90）可保护和恢复LPS介导的跨内皮高通透性。Hsp 90的超乙酰化抑制其伴侣蛋白功能，表明HDAC在LPS介导的跨内皮渗透性中的作用。因此，我们研究了HDAC在人肺微血管内皮细胞（HLMVEC）中LPS介导的跨内皮渗透性中的作用。方法：使用Flour‐de‐lys测定试剂盒测量HDAC活性，并使用镀金细胞基质阻抗传感（ECIS）阵列测量HLMVEC屏障功能。结果如下：来自暴露于LPS（0.2、1或5EU/ml）的HLMVEC的全细胞裂解物显示与媒介物处理的细胞相比HDAC活性显著增加。细胞分级显示，LPS诱导HDAC活性在细胞质和核提取物。用PI 3 k/Akt抑制剂Ly 294002（5μM）或Hsp 90抑制剂AUY-922（2μM）预处理未抑制LPS诱导的HDAC活性，表明LPS刺激HDAC活性独立于Hsp 90或Akt。此外，用HDAC抑制剂（曲古抑菌素（TSA），1 μM或帕比司他（LBH），1 μM）预处理可保护HLMVEC单层免受LPS介导的高通透性，并通过破坏Hsp 90-CDC 37相互作用减弱Hsp 90伴侣蛋白功能。结论：LPS诱导HDAC活性，可能通过去乙酰化和激活Hsp 90伴侣功能促进跨内皮高通透性。（支持HL 093460和HL 101902）
Objective: Lipopolysaccharide causes disruption of the endothelial barrier and increases trans‐endothelial permeability. Inhibition of heat shock protein 90 (Hsp90) protects and restores LPS‐mediated trans‐endothelial hyper‐permeability. Hyper‐acetylation of Hsp90 suppresses its chaperone function suggesting a role of HDAC in LPS‐mediated trans‐endothelial permeability. Therefore, we investigated the role of HDAC in LPS‐mediated trans‐endothelial permeability in human lung microvascular endothelial cells (HLMVEC). Method: HDAC activity was measured using the Flour‐de‐lys assay kit and HLMVEC barrier function was measured using gold‐plated electric cell‐substrate impedance sensing (ECIS) arrays. Results: Whole cell lysates from HLMVEC exposed to LPS (0.2, 1 or 5 EU/ml) showed significant increase in HDAC activity compared to vehicle‐treated cells. Cell fractionation revealed that LPS induced HDAC activity in both the cytoplasmic and the nuclear extracts. This LPS‐induced HDAC activity was not inhibited by pre‐treatment with the PI3k/Akt inhibitor, Ly294002 (5µM) or the Hsp90 inhibitor AUY‐922 (2µM), indicating that LPS stimulates HDAC activity independently of Hsp90 or Akt. Additionally, pre‐treatment with either HDAC inhibitor (Trichostatin (TSA), 1 µM or Panobinostat (LBH), 1 µM) protected HLMVEC monolayers from LPS‐mediated hyper‐permeability and attenuated Hsp90 chaperone function by disrupting Hsp90‐CDC37 interaction. Conclusion: LPS induces HDAC activity to promote trans‐endothelial hyper‐permeability possibly via de‐acetylation and activation of the Hsp90 chaperone function. (Supported by HL093460 and HL101902)