Conclusion
Coculturing M1/M2 macrophages and GECs treated with AGEs can affect the mRNA expression in GECs. M2 macrophages can inhibit the Braf, Fgfr2, Map3k1, Mras, and p-ERK expression in the MAPK signaling pathway, which may be related to M2 macrophages protecting GECs from damage by AGEs.
Methods
We cocultured M1/M2 macrophages and GECs treated with AGEs. The mRNA expression profile was detected by transcriptome sequencing. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis was performed using the online tool Database for Annotation, Visualization, and Integrated Discovery (DAVID). The expression of differential genes was confirmed using droplet digital PCR and Western blot.
Results
The results showed that 1,337 genes were significantly changed in M2 cocultured with AGE-treated GECs compared to those of the M1 cocultured with AGE-treated GECs. The KEGG analysis results show that the differentially expressed genes are primarily involved in specific pathways, including the rat sarcoma signaling pathway and the mitogen-activated protein kinase (MAPK) signaling pathway. The expression of Braf, Fgfr2, Map3k1, and Mras, which are involved in the MAPK signaling pathway, was downregulated in cocultured M2 with AGE-treated GECs and was consistent with sequencing. Therefore, the cocultured M1/M2 macrophages with AGE-treated GECs can affect the expression of mRNA in GECs, and M2 macrophages via the MAPK signaling pathway may protect GECs from damage by AGEs.