Agarotetrol alleviates reflux esophagitis by regulating autophagy through the METTL14/FOXO3a pathway

Agarotetrol exhibits significant anti-inflammatory properties for treating reflux esophagitis (RE). Recent research has revealed that autophagy plays a protective role against esophageal inflammation through pathways involving FOXO3a and METTL14 expression.

To examine the autophagy-mediated effects of agarotetrol on RE.

We established a RE rat model through pyloric clip combined with gastric fundus ligation. We assessed agarotetrol's anti-inflammatory effects using HE staining, pH measurements, and ELISA. The impact of agarotetrol on the FOXO3a-induced autophagy pathway was investigated using qRT-PCR, Western blot and immunohistochemistry. We examined how agarotetrol regulates METTL14-mediated m6A modification of FOXO3a using qRT-PCR, Western blot, immunohistochemistry, and MeRIP-qPCR. Molecular docking and MST experiment confirmed direct agarotetrol-METTL14 binding. METTL14 silencing via AAV-sh-METTL14 injection verified agarotetrol's regulatory mechanism on autophagy in RE through the METTL14/FOXO3a signaling pathway.

Agarotetrol treatment significantly reduced esophageal mucosal damage and inflammatory factor levels in RE rats while increasing esophageal pH. The treatment enhanced the FOXO3a-induced autophagy pathway, evidenced by upregulation of the LC3B-II expression and downregulation of p62 expression. METTL14-mediated m6A modification of FOXO3a mRNA increased following agarotetrol treatment, explaining the mechanism of elevated FOXO3a expression. Molecular docking and MST experiment confirmed direct binding between agarotetrol and METTL14. In METTL14 knockdown rats, agarotetrol failed to promote m6A modification of FOXO3a mRNA, preventing activation of the FOXO3a-mediated autophagy pathway.

Agarotetrol enhances the m6A modification of FOXO3a mRNA by binding to METTL14, thereby upregulating FOXO3a expression and activating the autophagy pathway, ultimately exerting a protective effect against RE.