Neutrophil extracellular traps promote corneal neovascularization-induced by alkali burn

角膜新生血管 中性粒细胞胞外陷阱 体内 血管生成 PI3K/AKT/mTOR通路 体外 化学 脐静脉 角膜 新生血管 细胞生物学 分子生物学 药理学 炎症 细胞外 免疫学 信号转导 生物 生物化学 癌症研究 生物技术 神经科学
作者
Kelan Yuan,Jiao Zheng,Xiaodan Huang,Yue Zhang,Yu Han,Renjian Hu,Xiuming Jin
出处
期刊:International Immunopharmacology [Elsevier]
卷期号:88: 106902-106902 被引量:30
标识
DOI:10.1016/j.intimp.2020.106902
摘要

To investigate the effects of neutrophil extracellular traps (NETs) on angiogenesis in vitro and in vivo and the regulatory role of mammalian target of rapamycin (mTOR) activity in it. The regulatory role of mTOR in NETs formation was explored. In vitro, human neutrophils were pretreated with rapamycin. NETs formation was measured using immunofluorescence staining of NETs markers, SYTOX Green and PicoGreen after NaOH stimulation. In vivo, mice were treated with rapamycin, and NETs formation in cornea was measured using immunofluorescence staining 7 days after alkali burn. Then, the effects of NETs on angiogenesis were investigated. In vitro, human neutrophils were treated with DNase I or rapamycin. NETs were isolated after NaOH stimulation and the isolated NETs were co-culture with human umbilical vein endothelial cells (HUVECs). HUVECs migration, proliferation, and inflammatory activation were measured. In vivo, mice were injected subconjunctivally with supernatant containing NETs. Corneal neovascularization was visualized by immunofluorescence staining. NETs structures can be observed in NaOH-stimulated neutrophils and alkali-burned mouse cornea compared with normal group. Treated with rapamycin enhanced NETs formation in response to NaOH management compared with DMSO control in vitro and in vivo. NETs increased the migration, proliferation and inflammatory activation of HUVECs, and subconjunctival injection of NETs promoted inflammatory and angiogenic response in corneal alkali burn model. NETs formation can be triggered by NaOH stimulation. mTOR activity has a negative regulatory effect on NETs formation. NETs promoted angiogenic responses and inflammatory activation of HUVECs and increased corneal neovascularization and inflammatory response.
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