Integrin Beta 4E Promotes Endothelial Phenotypic Changes and Attenuates Lung Endothelial Cell Inflammatory Responses.
Integrin Beta 4E Promotes Endothelial Phenotypic Changes and Attenuates Lung Endothelial Cell Inflammatory Responses.
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整合素 Beta 4E 促进内皮表型变化并减弱肺内皮细胞炎症反应
DOI：
10.3389/fphys.2022.769325
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发表时间：
2022
影响因子：
4
通讯作者：
Cress AE
中科院分区：
文献类型：
--
作者：
Chen W;Gard JMC;Epshtein Y;Camp SM;Garcia JGN;Jacobson JR;Cress AE
我们先前报道了整合素β 4（ITGB 4）是辛伐他汀（一种3-羟基-3-甲基戊二酰辅酶A还原酶抑制剂）保护肺血管的重要介质。在这项研究中，我们报告增加内皮细胞（EC）的表达，特别是ITGB 4 E，ITGB 4 mRNA剪接变异体，辛伐他汀对EC蛋白表达和炎症反应的影响。在初始实验中，在免疫沉淀与ITGB 4相关的整合素α 6（ITGA 6）和Western印迹法检测全长ITGB 4和ITGB 4 E（其独特特征为截短的114个氨基酸胞质结构域）之前，使用辛伐他汀（5 μM，24 h）处理人肺动脉EC。这些实验证实了全长ITGB 4和ITGB 4 E的显著增加。为了研究单独增加的ITGB 4 E表达的作用，用ITGB 4 E或对照载体转染EC，并将细胞接种在含有基质胶的威尔斯孔中以评估对血管生成的作用或用于划痕测定以评估迁移。与对照组相比，在ITGB 4 E转染的EC中观察到血管生成和迁移减少。在单独的实验中，来自转染细胞的PCR和蛋白质印迹显示EC蛋白表达的显着变化与ITGB 4 E的增加相关，包括血小板内皮细胞粘附分子-1（PECAM-1）和血管内皮钙粘蛋白（VEGF-cadherin）的显著降低（VE-钙粘蛋白）以及E-钙粘蛋白和N-钙粘蛋白的表达增加，沿着Slug和Snail转录因子的表达增加，这些转录因子促进内皮细胞向内皮细胞转化。间充质转化（EndMT）。然后，我们研究了ITGB 4 E过表达对EC炎症反应的功能性影响，并观察到脂多糖（LPS）诱导的丝裂原活化蛋白激酶（MAPK）活化的显著减弱，包括细胞外信号调节激酶（ERK）和c-Jun N末端激酶（JNK）的磷酸化降低，以及炎性细胞因子的减少。（IL-6和IL-8），在LPS或过度循环拉伸（CS）后的EC培养基中表达。最后，EC表达增加的ITGB 4 E表现出凝血酶诱导的屏障破坏减少，通过跨内皮电阻测量。我们的数据支持由ITGB 4 E诱导的不同EC表型变化，其也与细胞炎症反应的减弱相关。这些发现暗示ITGB 4 E上调是他汀类药物保护肺EC的重要介质，并可能为急性肺损伤（ALI）患者或有急性肺损伤风险的患者带来新的治疗策略。
We previously reported integrin beta 4 (ITGB4) is an important mediator of lung vascular protection by simvastatin, a 3-hydroxy-3-methylglutaryl-coenzyme A-reductase inhibitor. In this study, we report increased endothelial cell (EC) expression specifically of ITGB4E, an ITGB4 mRNA splice variant, by simvastatin with effects on EC protein expression and inflammatory responses. In initial experiments, human pulmonary artery ECs were treated using simvastatin (5 μM, 24 h) prior to immunoprecipitation of integrin alpha 6 (ITGA6), which associates with ITGB4, and Western blotting for full-length ITGB4 and ITGB4E, uniquely characterized by a truncated 114 amino acid cytoplasmic domain. These experiments confirmed a significant increase in both full-length ITGB4 and ITGB4E. To investigate the effects of increased ITGB4E expression alone, ECs were transfected with ITGB4E or control vector, and cells were seeded in wells containing Matrigel to assess effects on angiogenesis or used for scratch assay to assess migration. Decreased angiogenesis and migration were observed in ITGB4E transfected ECs compared with controls. In separate experiments, PCR and Western blots from transfected cells demonstrated significant changes in EC protein expression associated with increased ITGB4E, including marked decreases in platelet endothelial cell adhesion molecule-1 (PECAM-1) and vascular endothelial-cadherin (VE-cadherin) as well as increased expression of E-cadherin and N-cadherin along with increased expression of the Slug and Snail transcription factors that promote endothelial-to-mesenchymal transition (EndMT). We, then, investigated the functional effects of ITGB4E overexpression on EC inflammatory responses and observed a significant attenuation of lipopolysaccharide (LPS)-induced mitogen-activated protein kinase (MAPK) activation, including decreased phosphorylation of both extracellular signal-regulated kinase (ERK) and c-Jun N-terminal kinase (JNK), as well as reduced inflammatory cytokines (IL-6 and IL-8), expressed in the media of EC after either LPS or excessive cyclic stretch (CS). Finally, EC expression-increased ITGB4E demonstrated decreased barrier disruption induced by thrombin as measured by transendothelial electrical resistance. Our data support distinct EC phenotypic changes induced by ITGB4E that are also associated with an attenuation of cellular inflammatory responses. These findings implicate ITGB4E upregulation as an important mediator of lung EC protection by statins and may lead to novel therapeutic strategies for patients with or at risk for acute lung injury (ALI).
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DOI：
10.1083/jcb.201701176
发表时间：
2017-11-06
期刊：
The Journal of cell biology
影响因子：
--
作者：
Böttcher RT;Veelders M;Rombaut P;Faix J;Theodosiou M;Stradal TE;Rottner K;Zent R;Herzog F;Fässler R
通讯作者：
Fässler R
影响因子：
3.4
作者：
Li, Guo-Sheng;Hou, Wei;Wei, Hong-Yu
通讯作者：
Wei, Hong-Yu
影响因子：
3.4
作者：
Meng, Xiangli;Liu, Peng;Tan, Xiaodong
通讯作者：
Tan, Xiaodong
影响因子：
4
作者：
Bolós, V;Peinado, H;Cano, A
通讯作者：
Cano, A
影响因子：
11.2
作者：
Eastham, Angela M.;Spencer, Helen;Ward, Christopher M.
通讯作者：
Ward, Christopher M.