Inhibition of Pyk2 blocks lung inflammation and injury in a mouse model of acute lung injury.
Inhibition of Pyk2 blocks lung inflammation and injury in a mouse model of acute lung injury.
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DOI：
10.1186/1465-9921-13-4
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发表时间：
2012-01-18
影响因子：
5.8
通讯作者：
Zhu X
中科院分区：
文献类型：
--
作者：
Duan Y;Learoyd J;Meliton AY;Leff AR;Zhu X
富含脯氨酸的酪氨酸激酶 2 (Pyk2) 对于中性粒细胞脱颗粒和体外趋化作用至关重要。然而，其对LPS引起的急性肺损伤（ALI）期间肺部炎症和水肿形成过程的影响仍不清楚。本研究的目的是确定抑制 Pyk2 对 LPS 诱导的体内急性肺部炎症和损伤的影响。 C57BL6 小鼠气管内给予 10 mg/kg LPS 或盐水。 Pyk2 的抑制是通过在攻击前 1 小时腹膜内施用 TAT-Pyk2-CT 来实现的。 LPS 治疗后 18 小时对支气管肺泡灌洗液中的细胞计数、肺组织学和 BAL 中的蛋白质浓度进行分析。通过小鼠细胞因子多重试剂盒测量 BAL 中的 KC 和 MIP-2 浓度。使用计算机控制的小动物呼吸机通过压力-容量曲线确定静态肺顺应性。通过分光光度法测定肺匀浆中外渗的伊文思蓝浓度。气管内滴注 LPS 诱导显着的中性粒细胞浸润至肺间质和肺泡腔，并且通过 TAT-Pyk2-CT 预处理减弱了这种浸润。 TAT-Pyk2-CT 预处理还减弱了 1) 肺组织中的髓过氧化物酶含量，2) 通过肺部伊文思蓝染料外渗测量的血管渗漏和支气管肺泡灌洗液中蛋白质浓度的增加，以及 3) 肺顺应性的降低。在每个范例中，用对照蛋白 TAT-GFP 处理没有阻断效果。相比之下，TAT-Pyk2-CT 并未减少支气管肺泡灌洗液中中性粒细胞趋化因子 MIP-2 和角质细胞衍生趋化因子的产生。蛋白质印迹分析证实，通过 TAT-Pyk2-CT 预处理，LPS 攻击的肺中 Pyk2 的酪氨酸磷酸化降低至对照水平。这些结果表明，Pyk2 在小鼠急性肺损伤的发生过程中发挥着重要作用，Pyk2 的药理学抑制可能为面临急性肺损伤风险的患者的预处理提供潜在的治疗策略。
Proline-rich tyrosine kinase 2 (Pyk2) is essential in neutrophil degranulation and chemotaxis in vitro. However, its effect on the process of lung inflammation and edema formation during LPS induced acute lung injury (ALI) remains unknown. The goal of the present study was to determine the effect of inhibiting Pyk2 on LPS-induced acute lung inflammation and injury in vivo. C57BL6 mice were given either 10 mg/kg LPS or saline intratracheally. Inhibition of Pyk2 was effected by intraperitoneal administration TAT-Pyk2-CT 1 h before challenge. Bronchoalveolar lavage analysis of cell counts, lung histology and protein concentration in BAL were analyzed at 18 h after LPS treatment. KC and MIP-2 concentrations in BAL were measured by a mouse cytokine multiplex kit. The static lung compliance was determined by pressure-volume curve using a computer-controlled small animal ventilator. The extravasated Evans blue concentration in lung homogenate was determined spectrophotometrically. Intratracheal instillation of LPS induced significant neutrophil infiltration into the lung interstitium and alveolar space, which was attenuated by pre-treatment with TAT-Pyk2-CT. TAT-Pyk2-CT pretreatment also attenuated 1) myeloperoxidase content in lung tissues, 2) vascular leakage as measured by Evans blue dye extravasation in the lungs and the increase in protein concentration in bronchoalveolar lavage, and 3) the decrease in lung compliance. In each paradigm, treatment with control protein TAT-GFP had no blocking effect. By contrast, production of neutrophil chemokines MIP-2 and keratinocyte-derived chemokine in the bronchoalveolar lavage was not reduced by TAT-Pyk2-CT. Western blot analysis confirmed that tyrosine phosphorylation of Pyk2 in LPS-challenged lungs was reduced to control levels by TAT-Pyk2-CT pretreatment. These results suggest that Pyk2 plays an important role in the development of acute lung injury in mice and that pharmacological inhibition of Pyk2 might provide a potential therapeutic strategy in the pretreatment for patients at imminent risk of developing acute lung injury.
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DOI：
10.1083/jcb.200907135
发表时间：
2010-03-22
期刊：
The Journal of cell biology
影响因子：
--
作者：
Cain RJ;Vanhaesebroeck B;Ridley AJ
通讯作者：
Ridley AJ
影响因子：
6.4
作者：
Di Cioccio, V;Strippoli, R;Bertini, R
通讯作者：
Bertini, R
影响因子：
56.9
作者：
NOVOGRODSKY, A;VANICHKIN, A;LEVITZKI, A
通讯作者：
LEVITZKI, A
影响因子：
30.5
作者：
Guinamard, R;Okigaki, M;Ravetch, JV
通讯作者：
Ravetch, JV
影响因子：
5.7
作者：
Loftus, Joseph C.;Yang, Zhongbo;Lipinski, Christopher A.
通讯作者：
Lipinski, Christopher A.