Screening and functional validation of key genes in Helicobacter Pylori-induced macrophage M1 polarization: role in migraine-associated functional dyspepsia

BackgroundMigraine, a prevalent and debilitating neurovascular disorder, frequently co-occurs with functional dyspepsia. Emerging evidence suggests that Helicobacter pylori (H.

pylori) infection may contribute to both conditions via neuroimmune pathways, though the molecular basis for this association has yet to be fully elucidated.MethodsH. pylori-related datasets from GEO were analyzed to identify differentially expressed genes (DEGs), followed by functional enrichment analyses including GO, KEGG, GSEA, and GSVA.

Immune infiltration patterns were assessed, and CGRP-related hub genes were identified using machine learning approaches. The relationship between these hub genes and immune cell infiltration—particularly M1 macrophage polarization—was explored, and the interaction between CALCA and PNOC was analyzed by correlation analysis and protein-protein interaction networks.

Experimental validation was performed using RT-qPCR in H. pylori-infected THP-1 macrophages and SH-SY5Y neuronal cells, along with loss- and gain-of-function experiments to investigate regulatory relationships.ResultsTranscriptomic analysis of H.

pylori-infected patients identified 683 differentially expressed genes enriched in immune and inflammatory pathways. Immune profiling further revealed prominent M1 macrophage polarization with increased γδT cells and B lymphocytes.