miR-375-3p 通过靶向 Notch1 在低氧条件下调节大鼠肺微血管内皮细胞的活性。
MiR-375-3p regulates rat pulmonary microvascular endothelial cell activity by targeting Notch1 during hypoxia.
机构信息
Tianjin Medical University, Tianjin, P. R. China.
Qinhuangdao Haigang Hospital, Heibei, P. R. China.
出版信息
J Int Med Res. 2020 Jul;48(7):300060520926851. doi: 10.1177/0300060520926851.
OBJECTIVE
Pulmonary microvascular endothelial cells (PMECs) exhibit specific responses in adaptation to hypoxia. However, the mechanisms regulating PMEC activities during hypoxia remain unclear. This study investigated the potential involvement of a microRNA, miR-375-3p, in the regulation of PMEC activities.
METHODS
Primary PMECs were isolated from rats. The expression levels of miR-375-3p and Notch1 in the PMECs were detected by quantitative PCR and western blotting. Luciferase reporter assays were performed to explore the transcriptional regulation of Notch1 by miR-375-3p. The proliferation and chemotaxis of the PMECs were measured with the Cell Counting Kit-8 and Transwell invasion assays, respectively. Additionally, the capacity of hypoxia-treated PMECs for angiogenesis and inflammatory response was determined with tube formation assays and ELISA, respectively.
RESULTS
The expression of miR-375-3p and Notch1 in the PMECs was significantly down-regulated and up-regulated during hypoxia, respectively. The results demonstrated that miR-375-3p directly targets Notch1 in PMECs, thereby suppressing the transcriptional expression of . It was further revealed that miR-375-3p regulates the proliferation, chemotaxis, angiogenesis, and inflammatory response of PMECs.
CONCLUSIONS
Our findings revealed the important role of miR-375-3p in the regulation of PMEC function and suggest the potential involvement of miR-375-3p in the development of lung diseases.
目的
肺微血管内皮细胞（PMECs）在适应低氧环境时表现出特定的反应。然而，调节 PMEC 活动的机制在低氧环境下仍不清楚。本研究探讨了 microRNA miR-375-3p 在调节 PMEC 活性中的潜在作用。
方法
从大鼠中分离出原代 PMECs。通过定量 PCR 和 Western blot 检测 PMEC 中 miR-375-3p 和 Notch1 的表达水平。通过荧光素酶报告基因实验探讨 miR-375-3p 对 Notch1 的转录调控作用。通过细胞计数试剂盒-8 和 Transwell 侵袭实验分别测量 PMEC 的增殖和趋化性。此外，通过管形成实验和 ELISA 分别测定缺氧处理的 PMEC 的血管生成和炎症反应能力。
结果
在低氧环境中，PMEC 中 miR-375-3p 的表达明显下调，而 Notch1 的表达明显上调。结果表明，miR-375-3p 可直接靶向 PMECs 中的 Notch1，从而抑制 Notch1 的转录表达。进一步研究表明，miR-375-3p 调节 PMEC 的增殖、趋化、血管生成和炎症反应。
结论
本研究揭示了 miR-375-3p 在调节 PMEC 功能中的重要作用，并提示 miR-375-3p 可能参与肺部疾病的发生。
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