College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, China.
College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, China.
J Hazard Mater. 2021 Sep 15;418:126172. doi: 10.1016/j.jhazmat.2021.126172. Epub 2021 May 21.
Hydrogen sulfide (HS) is an air pollutant to cause tracheal injury. Pyroptosis is responsible for tissue injury through reactive oxygen species (ROS) production. Competitive endogenous RNAs (ceRNAs) chelate microRNAs and reduce their inhibitory effect on other transcripts, thus affecting ROS levels and pyroptosis. However, it is not clear how HS regulates pyroptosis via the ceRNA axis. Therefore, we established a broilers model of HS exposure for 42 days to assess pyroptosis and obtain a ceRNA network by immunohistochemistry and RNA sequencing. We detected pyroptosis induced by HS and verified circRNA-IGLL1-17828/miR-6631-5p/DUSP6 axis by a double luciferase reporter assay. We also measured ROS levels and the expression of pyroptotic indicators such as (Caspase1) Casp-1, Interleukin 1β (IL-1β) and Interleukin 1β (IL-18). miR-6631-5p knockdown decreased pyroptotic indicators induced by HS. Overexpression of miR-6631-5p or DUSP6 knockdown stimulated ROS generation and upregulated pyroptotic indicators. N-acetyl--cysteine (NAC) decreased pyroptotic indicators and ROS levels both induced by miR-6631-5p. Moreover, circRNA-IGLL1-17828, participated in intermolecular competition as a ceRNA of DUSP6. In conclusion, circRNA-IGLL1-17828/miR-6631-5p/DUSP6 crosstalk regulated HS-induced pyroptosis in broilers trachea via ROS generation. This is the first study to reveal regulation mechanism of circRNA-related CeRNAs on pyroptosis induced by HS, providing important reference for environmental toxicology.
硫化氢（HS）是一种引起气管损伤的空气污染物。细胞焦亡通过活性氧（ROS）的产生导致组织损伤。竞争性内源性 RNA（ceRNA）螯合 microRNA，并减少其对其他转录物的抑制作用，从而影响 ROS 水平和细胞焦亡。然而，HS 如何通过 ceRNA 轴调节细胞焦亡尚不清楚。因此，我们建立了一个暴露于 HS 的肉鸡模型 42 天，以评估细胞焦亡，并通过免疫组织化学和 RNA 测序获得 ceRNA 网络。我们检测了 HS 诱导的细胞焦亡，并通过双荧光素酶报告基因实验验证了 circRNA-IGLL1-17828/miR-6631-5p/DUSP6 轴。我们还测量了 ROS 水平和细胞焦亡标志物（Caspase1） Casp-1、白细胞介素 1β（IL-1β）和白细胞介素 18（IL-18）的表达。miR-6631-5p 的敲低降低了 HS 诱导的细胞焦亡标志物。miR-6631-5p 的过表达或 DUSP6 的敲低刺激了 ROS 的产生并上调了细胞焦亡标志物。N-乙酰-L-半胱氨酸（NAC）降低了 miR-6631-5p 诱导的细胞焦亡标志物和 ROS 水平。此外，circRNA-IGLL1-17828 作为 DUSP6 的 ceRNA 参与了分子间竞争。总之，circRNA-IGLL1-17828/miR-6631-5p/DUSP6 串扰通过 ROS 的产生调节了肉鸡气管中 HS 诱导的细胞焦亡。这是首次揭示 circRNA 相关 CeRNA 对 HS 诱导的细胞焦亡的调节机制的研究，为环境毒理学提供了重要参考。
