食用油油烟衍生的 PM 通过 ROS 介导的 NLRP3 炎性小体通路对人脐静脉内皮细胞血管生成的影响。
The effects of cooking oil fumes-derived PM on blood vessel formation through ROS-mediated NLRP3 inflammasome pathway in human umbilical vein endothelial cells.
机构信息
Department of Occupational Health and Environmental Health, School of Public Health, Anhui Medical University, 81 Meishan Road, Hefei, Anhui 230032, China.
Department of Environmental Health, Center for Disease Control and Prevention, Nanjing, China.
出版信息
Ecotoxicol Environ Saf. 2019 Jun 15;174:690-698. doi: 10.1016/j.ecoenv.2019.03.028. Epub 2019 Mar 14.
BACKGROUND
Cooking oil fumes (COFs), a main pollutant in kitchen air, is a major risk to human health. In our previous research, exposure to COFs-derived PM could cause umbilical vascular endothelial dysfunction, leading to decreased fetal weight. Here, to test the role of ROS-mediated NLRP3 inflammasome pathway in blood vessel formation of human umbilical vein endothelial cells (HUVECs) caused by COFs-derived PM, the cells were exposed to COFs-derived PM at different concentrations with and without N-acetyl-L-cysteine (NAC).
METHODS
MTT assay was used to determine HUVECs viability. Intracellular ROS and mitochondrial ROS levels were assessed with DCFH-DA and MitoSOX™ assay. The levels of proteins and mRNA involved in NLRP3 inflammasome signaling pathway and VEGF were measured by western blot and real-time PCR (RT-PCR). Tube formation in HUVECs was detected by tube formation assay.
RESULTS
The results revealed that COFs-derived PM exposure reduced HUVECs viability, increased the intracellular and mitochondrial ROS levels in cells, and up-regulated the levels of proteins and mRNA involved in NLRP3 inflammasome signaling pathway. However, the protein and mRNA expression of VEGF were reduced with the increasing exposure concentrations. In addition, COFs-derived PM also affected the tube formation. However, co-incubation with NAC effectively rescued the damages caused by COFs-derived PM exposure.
CONCLUSIONS
This study proved that COFs-derived PM could significantly reduce HUVECs viability, induce the overproduction of ROS, lead to inflammation and inhibit VEGF expression, thus affect angiogenesis of HUVECs in vitro. It was revealed that the impact caused by COFs-derived PM on blood vessel formation through a ROS-mediated NLRP3 inflammasome pathway.
背景
食用油烟雾（COFs）是厨房空气中的主要污染物，也是人类健康的主要风险因素。在我们之前的研究中，暴露于 COFs 衍生的 PM 可导致脐血管内皮功能障碍，导致胎儿体重下降。在这里，为了测试 COFs 衍生的 PM 引起的人脐静脉内皮细胞（HUVEC）血管形成中 ROS 介导的 NLRP3 炎性小体途径的作用，用不同浓度的 COFs 衍生的 PM 以及 N-乙酰-L-半胱氨酸（NAC）处理细胞。
方法
MTT 法测定 HUVECs 活力。用 DCFH-DA 和 MitoSOXTM 测定细胞内 ROS 和线粒体 ROS 水平。用 Western blot 和实时 PCR（RT-PCR）测定 NLRP3 炎性小体信号通路和 VEGF 相关蛋白和 mRNA 的水平。用管形成试验检测 HUVECs 的管形成。
结果
结果表明，COFs 衍生的 PM 暴露降低了 HUVECs 的活力，增加了细胞内和线粒体 ROS 水平，并上调了 NLRP3 炎性小体信号通路相关蛋白和 mRNA 的水平。然而，随着暴露浓度的增加，VEGF 的蛋白和 mRNA 表达减少。此外，COFs 衍生的 PM 还影响管形成。然而，用 NAC 共孵育可有效挽救 COFs 衍生的 PM 暴露引起的损伤。
结论
本研究证明 COFs 衍生的 PM 可显著降低 HUVECs 的活力，诱导 ROS 过度产生，导致炎症和抑制 VEGF 表达，从而影响 HUVECs 的体外血管生成。揭示了 COFs 衍生的 PM 通过 ROS 介导的 NLRP3 炎性小体途径对血管形成的影响。
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