Institute of Genetics and Regenerative Biology, College of Life Sciences, Zhejiang University, Hangzhou, Zhejiang, China.
Research Center for Air Pollution and Health, Zhejiang University, Hangzhou, Zhejiang, China.
Br J Pharmacol. 2018 Apr;175(8):1241-1259. doi: 10.1111/bph.14036. Epub 2017 Oct 25.
Cardiovascular disease associated with antiretroviral therapy (ART) has become a major clinical challenge for HIV-positive patients. However, the role of ART in blood vessel growth is largely unknown. Here, we examined an integral component of ART, nucleoside/nucleotide reverse transcriptase inhibitors (NRTIs) and investigated their effects on key microvascular functions, including angiogenesis and lymphangiogenesis.
The angiogenesis/lymphangiogenesis capability of endothelial cells (ECs) was evaluated using migration, proliferation and tube formation assays in vitro, and mouse ear and Matrigel plug assays in vivo. Expressions of signalling molecules and mitochondrial antioxidant catalases were determined using Western blotting. Receptor tyrosine kinase (RTK) internalization and endocytosis were examined using flow cytometry and confocal immunofluorescence microscopy respectively. Mitochondrial DNA copy number and ROS were determined using quantitative real-time PCR and MitoSOX staining respectively.
Pharmaceutical doses of NRTIs [azidothymidine (AZT), tenofovir disoproxil fumarate (TDF) and lamivudine (3TC)] inhibited angiogenesis and lymphangiogenesis both in vivo and in vitro by affecting the proliferation and migration of ECs. Correspondingly, NRTIs selectively attenuated the activation and transduction of endothelial RTK signals, VEGFR2 and FGFR1 pathways, in vascular ECs and the VEGFR3 pathway in lymphatic ECs. Both TDF and 3TC restrained RTKs' endocytosis into early endosomes but not internalization, while AZT blocked the protein maturation of RTKs. Excessive ROS levels were detected in NRTI-treated ECs, and the MnSOD mimic MnTMPyP alleviated the angiogenic/lymphangiogenic defects induced by NRTIs.
NRTIs negatively regulate angiogenesis and lymphangiogenesis by inducing mitochondrial oxidative stress and subsequently impairing RTK signalling in ECs.
This article is part of a themed section on Spotlight on Small Molecules in Cardiovascular Diseases. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v175.8/issuetoc.
与抗逆转录病毒疗法（ART）相关的心血管疾病已成为 HIV 阳性患者的主要临床挑战。然而，ART 在血管生成中的作用在很大程度上尚不清楚。在这里，我们研究了 ART 的一个组成部分，即核苷/核苷酸逆转录酶抑制剂（NRTIs），并研究了它们对关键微血管功能（包括血管生成和淋巴管生成）的影响。
使用体外迁移、增殖和管形成测定法以及体内小鼠耳和 Matrigel plugs 测定法评估内皮细胞（ECs）的血管生成/淋巴管生成能力。使用 Western blot 测定法测定信号分子和线粒体抗氧化剂过氧化氢酶的表达。使用流式细胞术和共聚焦免疫荧光显微镜分别检查受体酪氨酸激酶（RTK）内化和内吞作用。使用定量实时 PCR 和 MitoSOX 染色分别测定线粒体 DNA 拷贝数和 ROS。
药物剂量的 NRTIs[叠氮胸苷（AZT）、富马酸替诺福韦二吡呋酯（TDF）和拉米夫定（3TC）]通过影响 ECs 的增殖和迁移，在体内和体外均抑制血管生成和淋巴管生成。相应地，NRTIs 选择性地减弱了血管内皮 RTK 信号、VEGFR2 和 FGFR1 途径以及淋巴管内皮 VEGFR3 途径的激活和转导。TDF 和 3TC 均抑制 RTK 进入早期内体，但不抑制内化，而 AZT 阻断了 RTK 的蛋白成熟。在 NRTI 处理的 ECs 中检测到过多的 ROS 水平，MnSOD 模拟物 MnTMPyP 减轻了 NRTI 诱导的血管生成/淋巴管生成缺陷。
NRTIs 通过诱导线粒体氧化应激，随后损害 ECs 中的 RTK 信号，从而负调节血管生成和淋巴管生成。
本文是心血管疾病中小分子聚焦专题的一部分。要查看本部分中的其他文章，请访问 http://onlinelibrary.wiley.com/doi/10.1111/bph.v175.8/issuetoc。
