Hydrogen sulfide exposure induces pyroptosis in the trachea of broilers via the regulatory effect of circRNA-17828/miR-6631-5p/DUSP6 crosstalk on ROS production

Hydrogen sulfide (H2S) 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 H2S regulates pyroptosis via the ceRNA axis. Therefore, we established a broilers model of H2S exposure for 42 days to assess pyroptosis and obtain a ceRNA network by immunohistochemistry and RNA sequencing. We detected pyroptosis induced by H2S 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 H2S. Overexpression of miR-6631-5p or DUSP6 knockdown stimulated ROS generation and upregulated pyroptotic indicators. N-acetyl-L-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 H2S-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 H2S, providing important reference for environmental toxicology.