Supplementary Material for: Melatonin regulates lncRNA NEAT1/miR-138-5p/HIF-1α axis through MOV10 to affect acid-related esophageal epithelial cell pyroptosis

Introduction: Acid-related inflammatory damage to the esophageal epithelium is a key component in the development of gastroesophageal reflux disease (GERD). Melatonin (MT) is considered as a potential therapeutic agent, but its molecular mechanism is unknown. Methods: The expression of HIF-1α and pyroptosis-related genes (NLRP3, Caspase-1, IL-1β, and IL-18) was analyzed using bioinformatics methods in GSE63401 and validated using qRT-PCR and WB in a HEEC inflammation model induced by deoxycholic acid (DCA). Hoechst 33342/PI double staining was used to assess the level of pyroptosis, and the effect of MT treatment was observed. The miRDB, TarBase, miRcode, miRNet, and ENCORI databases were used to predict the lncRNA targeting HIF-1α and the RNA-binding protein (RBP) interacting with the lncRNA. Results: The expressions of MOV10, lncRNA NEAT1, HIF-1α, and pyroptosis-related genes were up-regulated, while the expression of miR-138-5p was down-regulated in acidic DCA-induced HEEC inflammation. MOV10 may bind to lncRNA NEAT1 and stabilize its expression, while lncRNA NEAT1 up-regulates the expression of HIF-1α by adsorbing miR-138-5p to activate the NLRP3 inflammasome. However, MT pretreatment can significantly inhibit these processes. Conclusions: MOV10-lncRNA NEAT1/ miR-138-5p/ HIF-1α/ NLRP3 axis plays a crucial role in acid-related esophageal epithelial inflammatory injury, and MT may exert an esophageal protective effect by inhibiting the pathway.