Department of Cardiothoracic Surgery, Children's Hospital of Chongqing Medical University, Chongqing, China; Pediatric Research Institute, Chongqing Key Laboratory of Pediatrics, Ministry of Education Key Laboratory of Child Development and Disorders, National Clinical Research Center for Child Health and Disorders, Children's Hospital of Chongqing Medical University, China.
The Second Affiliated Hospital of Chongqing Medical University, Chongqing, China.
J Adv Res. 2024 Feb;56:31-41. doi: 10.1016/j.jare.2023.03.003. Epub 2023 Mar 17.
It has been shown that polystyrenenanoplastic (PS-NP) exposure induces toxicity in the lungs.
This study aims to provide foundational evidence to corroborate that ferroptosis and abnormal HIF-1α activity are the main factors contributing to pulmonary dysfunction induced by PS-NP exposure.
Fifty male and female C57BL/6 mice were exposed to distilled water or 100 nm or 200 nm PS-NPs via intratracheal instillation for 7 consecutive days. Hematoxylin and eosin (H&E) and Masson trichrome staining were performed to observe the histomorphological changes in the lungs. To clarify the mechanisms of PS-NP-induced lung injury, we used 100 μg/ml, 200 μg/ml and 400 μg/ml 100 or 200 nm PS-NPs to treat the human lung bronchial epithelial cell line BEAS-2B for 24 h. RNA sequencing (RNA-seq) of BEAS-2B cells was performed following exposure. The levels of glutathione, malondialdehyde, ferrous iron (Fe), and reactive oxygen species (ROS) were measured. The expression levels of ferroptotic proteins were detected in BEAS-2B cells and lung tissues by Western blotting. Western blotting, immunohistochemistry, and immunofluorescence were used to evaluate the HIF-1α/HO-1 signaling pathway activity.
H&E staining revealed substantial perivascular lymphocytic inflammation in a bronchiolocentric pattern, and Masson trichrome staining demonstrated critical collagen deposits in the lungs after PS-NP exposure. RNA-seq revealed that the differentially expressed genes in PS-NP-exposed BEAS-2B cells were enriched in lipid metabolism and iron ion binding processes. After PS-NP exposure, the levels of malondialdehyde, Fe, and ROS were increased, but glutathione level was decreased. The expression levels of ferroptotic proteins were altered significantly. These results verified that PS-NP exposure led to pulmonary injury through ferroptosis. Finally, we discovered that the HIF-1α/HO-1 signaling pathway played an important role in regulating ferroptosis in the PS-NP-exposed lung injury.
PS-NP exposure caused ferroptosis in bronchial epithelial cells by activating the HIF-1α/HO-1 signaling pathway, and eventually led to lung injury.
已有研究表明，聚苯乙烯纳米塑料（PS-NP）暴露会导致肺部毒性。
本研究旨在提供基础证据，以证实铁死亡和异常 HIF-1α 活性是 PS-NP 暴露引起肺功能障碍的主要因素。
50 只雄性和雌性 C57BL/6 小鼠通过气管内滴注分别暴露于去离子水或 100nm 或 200nm PS-NP 中，连续 7 天。进行苏木精和伊红（H&E）和 Masson 三色染色以观察肺部的组织形态学变化。为了阐明 PS-NP 诱导的肺损伤机制，我们使用 100μg/ml、200μg/ml 和 400μg/ml 的 100nm 或 200nm PS-NP 处理人肺支气管上皮细胞系 BEAS-2B 24 小时。暴露后对 BEAS-2B 细胞进行 RNA 测序（RNA-seq）。测量谷胱甘肽、丙二醛、二价铁（Fe）和活性氧（ROS）的水平。通过 Western blot 检测 BEAS-2B 细胞和肺组织中铁死亡蛋白的表达水平。Western blot、免疫组织化学和免疫荧光用于评估 HIF-1α/HO-1 信号通路活性。
H&E 染色显示 PS-NP 暴露后肺部出现以细支气管为中心的血管周围淋巴细胞炎症，Masson 三色染色显示肺部有重要的胶原沉积。RNA-seq 显示 PS-NP 暴露的 BEAS-2B 细胞中差异表达的基因富集在脂质代谢和铁离子结合过程中。PS-NP 暴露后，丙二醛、Fe 和 ROS 的水平增加，而谷胱甘肽水平降低。铁死亡蛋白的表达水平发生显著改变。这些结果证实 PS-NP 暴露通过铁死亡导致肺损伤。最后，我们发现 HIF-1α/HO-1 信号通路在调节 PS-NP 暴露引起的肺损伤中铁死亡中起重要作用。
PS-NP 暴露通过激活 HIF-1α/HO-1 信号通路导致支气管上皮细胞中铁死亡，最终导致肺损伤。
