Reduced intracellular chloride concentration impairs angiogenesis by inhibiting oxidative stress-mediated VEGFR2 activation
Reduced intracellular chloride concentration impairs angiogenesis by inhibiting oxidative stress-mediated VEGFR2 activation
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细胞内氯浓度降低通过抑制氧化应激介导的 VEGFR2 激活来损害血管生成
DOI：
10.1038/s41401-020-0458-7
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发表时间：
2020-07-21
影响因子：
8.2
通讯作者：
Shang, Jin-yan
中科院分区：
文献类型：
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作者：
Li, Kai;Liu, Ying-ying;Shang, Jin-yan
氯离子（Cl-）稳态在心血管系统中具有重要意义。血清Cl（-）水平与心力衰竭患者的死亡率呈负相关。考虑到血管生成在心衰过程中的重要性，本研究旨在探讨细胞内Cl（-）浓度降低（[Cl-](i)）是否以及如何影响血管生成。用正常Cl（-）培养基或低Cl（-）培养基处理人脐内皮细胞（HUVECs）。我们发现[Cl-](i)的减少（从33.2到16.18 mM）抑制HUVEC增殖、迁移、细胞骨架重组、管形成，并随后抑制血管生成，在基础条件下，VEGF刺激或缺氧处理。此外，在低Cl（-）培养基中，vegf诱导的nadph介导的活性氧（ROS）的产生和VEGFR2轴的激活明显减弱。我们发现，降低[Cl-](i)抑制了NADPH的膜结合催化亚基p22phox和Nox2的表达，并减弱了细胞质调节亚基p47phox和p67phox的易位，从而限制了NADPH氧化酶复合物的形成和激活。此外，[Cl-]的降低(i)增强了ros相关蛋白酪氨酸磷酸酶1B （PTP1B）的活性，并增加了VEGFR2和PTP1B的相互作用。药理抑制PTP1B逆转了降低[Cl-](i)对VEGFR2磷酸化和血管生成的影响。在小鼠后肢缺血模型中，使用Cl（-）通道抑制剂DIDS或DCPIB （10 mg/kg, ig，每隔一天，连续2周）阻断Cl（-）外排可显著促进血流恢复和新毛细血管形成。总之，[Cl-](i)的减少通过抑制氧化酶应激介导的VEGFR2信号激活，从而抑制NADPH氧化酶复合物的形成和促进VEGFR2/PTP1B的关联，从而抑制血管生成，表明[Cl-](i)的调节可能是治疗血管生成功能障碍相关疾病的一种新的治疗途径。
Chloride (Cl-) homeostasis is of great significance in cardiovascular system. Serum Cl(-)level is inversely associated with the mortality of patients with heart failure. Considering the importance of angiogenesis in the progress of heart failure, this study aims to investigate whether and how reduced intracellular Cl(-)concentration ([Cl-](i)) affects angiogenesis. Human umbilical endothelial cells (HUVECs) were treated with normal Cl(-)medium or low Cl(-)medium. We showed that reduction of [Cl-](i)(from 33.2 to 16.18 mM) inhibited HUVEC proliferation, migration, cytoskeleton reorganization, tube formation, and subsequently suppressed angiogenesis under basal condition, and VEGF stimulation or hypoxia treatment. Moreover, VEGF-induced NADPH-mediated reactive oxygen species (ROS) generation and VEGFR2 axis activation were markedly attenuated in low Cl(-)medium. We revealed that lowering [Cl-](i)inhibited the expression of the membrane-bound catalytic subunits of NADPH, i.e., p22phox and Nox2, and blunted the translocation of cytosolic regulatory subunits p47phox and p67phox, thereby restricting NADPH oxidase complex formation and activation. Furthermore, reduced [Cl-](i)enhanced ROS-associated protein tyrosine phosphatase 1B (PTP1B) activity and increased the interaction of VEGFR2 and PTP1B. Pharmacological inhibition of PTP1B reversed the effect of lowering [Cl-](i)on VEGFR2 phosphorylation and angiogenesis. In mouse hind limb ischemia model, blockade of Cl(-)efflux using Cl(-)channel inhibitors DIDS or DCPIB (10 mg/kg, i.m., every other day for 2 weeks) significantly enhanced blood flow recovery and new capillaries formation. In conclusion, decrease of [Cl-](i)suppresses angiogenesis via inhibiting oxidase stress-mediated VEGFR2 signaling activation by preventing NADPH oxidase complex formation and promoting VEGFR2/PTP1B association, suggesting that modulation of [Cl-](i)may be a novel therapeutic avenue for the treatment of angiogenic dysfunction-associated diseases.