Epithelial cell-derived exosomes carry NamiRNA-143-5p and promote ASPN expression in fibroblasts to induce Helicobacter pylori infected gastritis progression

In Helicobacter pylori (H. pylori) associated gastritis, fibroblasts are recruited to inflammatory sites and secrete multiple pro-inflammatory cytokines, but the underlying mechanism remains unclear. Here, immunohistochemical staining revealed that ASPN expression was significantly upregulated in fibroblasts from H. pylori positive gastritis tissues, whereas its level remained unchanged in fibroblasts directly infected with H. pylori. Co-culture assays demonstrated that exosomes released from H. pylori infected epithelial cells induced the upregulation of ASPN and its downstream cytokines (IL-4, IL-6, and TGF-β) in fibroblasts. MicroRNA sequencing and correlation analyses identified miR-143-5p as an exosomal miRNA enriched after H. pylori infection that potentially regulates ASPN. Immunofluorescence confirmed that exosomes derived from epithelial cells carrying miR-143-5p were internalized by fibroblasts. Further immunofluorescence and immunohistochemical analyses showed nuclear accumulation of miR-143-5p in fibroblasts in both the H. pylori infected epithelial cell co-culture system and H. pylori positive gastritis tissues. Mechanistic studies demonstrated that miR-143-5p functions as a nuclear activating microRNA (NamiRNA-143-5p), binding to the super-enhancer region of ASPN, increasing H3K27ac enrichment, and promoting its transcription. In vivo, antagomir-143-5p reduced H. pylori induced ASPN and cytokine overexpression, thereby alleviating gastric inflammation. Thus, we conclude that exosomal NamiRNA-143-5p from H. pylori infected epithelial cells upregulate pro-inflammatory cytokines in fibroblasts by activating ASPN expression through a super-enhancer-dependent pathway, thereby positioning this axis as a potential therapeutic target for H. pylori associated gastric disease.