Lactobacillus johnsonii N5 Supernatant Protects RPMI-2650 Cells from HDM-Induced Injury in Association with Suppression of the NR4A1–IRE1–XBP1-Mediated Unfolded Protein Response

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Background/Objectives: House dust mites (HDMs) are significant allergens that cause damage to nasal epithelial cells and have a role in the pathophysiology of allergic rhinitis. Lactobacillus johnsonii N5 supernatant (N5sup), a probiotic strain, exhibits possible protective properties; nevertheless, the molecular processes involved remain unclear. Methods: RPMI-2650 human nasal epithelial carcinoma cells were subjected to treatment with HDM, N5sup, or a combination of both. Results: The HDM stimulation induced a certain degree of transcriptional alterations. Compared with the control group, a total of 221 differentially expressed genes were identified in HDM-treated cells, which were mainly enriched in the IRE1–XBP1 branch of the unfolded protein response (UPR) pathway. Comprehensive investigation demonstrated that HDM increased the NR4A1–IRE1–XBP1 signaling pathway, characterized by elevated expression levels of NR4A1, ERN1 (IRE1), and XBP1. Both qRT-PCR and Western blotting verified that N5sup strongly inhibited the HDM-induced increase in NR4A1, IRE1, and XBP1. Analysis of the protein–protein interaction network further confirmed a central hub comprising NR4A1, IRE1, and XBP1. Conclusions: N5sup supernatant protects RPMI-2650 cells from HDM-induced injury, and this protective effect is associated with the NR4A1–IRE1–XBP1 pathway. These findings suggest that this signaling axis may serve as a potential therapeutic target for probiotic intervention in allergic airway diseases.

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