Das Jayanta K, Felty Quentin
Department of Environmental & Occupational Health, Florida International University, Miami, Florida, United States of America.
PLoS One. 2014 Aug 4;9(8):e104159. doi: 10.1371/journal.pone.0104159. eCollection 2014.
Microvascular lesions resulting from endothelial cell dysfunction are produced in the brain, lung, kidney, and retina of patients of complex chronic diseases. The environmental and molecular risk factors which may contribute in the development of microvascular damage are unclear. The mechanism(s) responsible for initiating microvascular damage remain poorly defined, although several inciting factors have been proposed, including environmental toxicants-induced oxidative stress. Enhanced neovascularization has been implicated in either the development or progression of proliferative vascular lesions. Here, we present evidence for how PCB-induced ROS may contribute to the development of a neovascular phenotype with the aim of elucidating the role of environmental toxicants in endothelial dysfunction with a specific focus on the inhibitor of differentiation protein ID3. We used a combination of phenotype and immunohistochemical analysis followed by validating with protein expression and post-translational modifications with Western Blot and MALDI-TOF/TOF analysis. We also looked for a correlation between ID3 expression in vascular tissue. Our results showed that PCB-induced ROS mediated a highly tube branched neovascular phenotype that also depended on ID3 and Pyk2; and PCB153 treatment increased the size of endothelial spheroids under conditions typically used for clonal selection of stem cell spheroids. High ID3 protein expression correlated with a greater degree of malignancy and oxidative DNA damage marker 8-OHdG in blood vessels from human subjects. PCB153 treatment increased both serine and tyrosine phosphorylation of endothelial ID3. Stable ID3 overexpression increased cell survival of human microvascular endothelial cell line hCMEC/D3. In summary, our data provide evidence that ID3 may play a critical role in regulating vascular endothelial cell survival and development of microvascular lesions induced by persistent environmental pollutants such as PCB153. Findings of this study are important because they provide a new paradigm by which PCBs may contribute to the growth of microvascular lesions.
复杂慢性疾病患者的大脑、肺、肾和视网膜中会产生由内皮细胞功能障碍导致的微血管病变。可能导致微血管损伤发生的环境和分子风险因素尚不清楚。尽管已经提出了几种诱发因素,包括环境毒物诱导的氧化应激,但引发微血管损伤的机制仍未明确界定。新生血管形成增强与增殖性血管病变的发生或进展有关。在此,我们提供证据证明多氯联苯诱导的活性氧如何可能促成新生血管表型的发展,目的是阐明环境毒物在内皮功能障碍中的作用,特别关注分化抑制蛋白ID3。我们结合了表型和免疫组化分析,随后通过蛋白质表达验证以及蛋白质印迹和基质辅助激光解吸电离飞行时间/飞行时间分析进行翻译后修饰分析。我们还研究了血管组织中ID3表达之间的相关性。我们的结果表明,多氯联苯诱导的活性氧介导了高度分支的新生血管表型,这也依赖于ID3和粘着斑激酶2;在通常用于干细胞球克隆选择的条件下,多氯联苯153处理增加了内皮球的大小。在人类受试者的血管中,高ID3蛋白表达与更高程度的恶性肿瘤和氧化DNA损伤标志物8-羟基脱氧鸟苷相关。多氯联苯153处理增加了内皮ID3的丝氨酸和酪氨酸磷酸化。稳定的ID3过表达增加了人微血管内皮细胞系hCMEC/D3的细胞存活率。总之,我们的数据提供了证据,表明ID3可能在调节血管内皮细胞存活以及由多氯联苯153等持久性环境污染物诱导的微血管病变发展中起关键作用。本研究的发现很重要,因为它们提供了一个新的范例,通过该范例多氯联苯可能促成微血管病变的生长。