Institute of Environmental Systems Biology, Dalian Maritime University, Linghai Road 1, Dalian, 116026, China.
State Key Laboratory of Environmental Chemistry and Ecotoxicology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Shuangqing Road 18, Beijing, 100085, China.
Ecotoxicol Environ Saf. 2020 Apr 1;192:110267. doi: 10.1016/j.ecoenv.2020.110267. Epub 2020 Feb 7.
Endosulfan is a persistent organic pollutant and can cause endothelial dysfunction, closely related to cardiovascular diseases. Endothelial cell migration plays a critical role in atherosclerosis and angiogenesis. This study was aimed to investigate the effect of environmentally relevant doses of endosulfan and underlying molecular mechanism on endothelial cell migration. Human umbilical vein endothelial cells (HUVECs) were treated with DMSO (control) or endosulfan (0.1, 1, 10 and 20 μM) in the presence or absence of inhibitors. Wound healing and Transwell assay were employed to explore the effect of endosulfan on endothelial cell migration. The expression of genes or proteins was assayed by real-time PCR or immunoblotting. The results showed that endosulfan at relative low concentration (0.1, 1, 10 and 20 μM) increased cell migration ability horizontally and vertically at 12 h after exposure. In line with this cellular effect, Protein-tyrosine Phosphatase 4A3 (PTP4A3) expression was significantly increased in endosulfan-exposed endothelial cells. Specific inhibitor of PTP4A3 significantly inhibited 20 μM endosulfan-induced cell migration, the expression and phosphorylation of Src and phosphorylation of focal adhesion kinase (FAK). Exposure to endosulfan resulted in activation of various signaling pathways including phosphoinositide 3-kinase (PI3K)/AKT, mitogen-activated protein kinase (MAPK) and nuclear factor kappa B (NF-κB), which were suppressed by PTP4A3 inhibitor or specific inhibitor for each signaling pathway. Exposure to endosulfan significantly reduced nitric oxide production and caused oxidative stress in HUVECs. These findings suggest that endosulfan promoted cell migration through PTP4A3-mediated various signaling pathways in endothelial cells.
硫丹是一种持久性有机污染物，可导致血管内皮功能障碍，与心血管疾病密切相关。内皮细胞迁移在动脉粥样硬化和血管生成中起着关键作用。本研究旨在探讨环境相关剂量的硫丹及其潜在的分子机制对内皮细胞迁移的影响。用 DMSO（对照）或硫丹（0.1、1、10 和 20 μM）处理人脐静脉内皮细胞（HUVEC），并在存在或不存在抑制剂的情况下进行处理。采用划痕愈合和 Transwell 实验研究硫丹对内皮细胞迁移的影响。通过实时 PCR 或免疫印迹法测定基因或蛋白质的表达。结果表明，硫丹在相对较低的浓度（0.1、1、10 和 20 μM）下，在暴露后 12 小时可增加细胞的水平和垂直迁移能力。与这种细胞效应一致，暴露于硫丹的内皮细胞中蛋白酪氨酸磷酸酶 4A3（PTP4A3）的表达显著增加。PTP4A3 的特异性抑制剂显著抑制了 20 μM 硫丹诱导的细胞迁移、Src 的表达和磷酸化以及粘着斑激酶（FAK）的磷酸化。硫丹暴露导致多种信号通路的激活，包括磷酸肌醇 3-激酶（PI3K）/AKT、丝裂原活化蛋白激酶（MAPK）和核因子 kappa B（NF-κB），这些信号通路可被 PTP4A3 抑制剂或每种信号通路的特异性抑制剂抑制。硫丹暴露显著降低了 HUVEC 中一氧化氮的产生并引起氧化应激。这些发现表明，硫丹通过内皮细胞中 PTP4A3 介导的多种信号通路促进细胞迁移。
