miR-155 promotes macrophage pyroptosis induced by Porphyromonas gingivalis through regulating the NLRP3 inflammasome

Abstract

Objective: The aim of this study is to detect pyroptosis in macrophages stimulated with Porphyromonas gingivalis and elucidate the mechanism by which P. gingivalis induces pyroptosis in macrophages.

Methods: The immortalized human monocyte cell line U937 was stimulated with P. gingivalis W83. Flow cytometry was carried out to detect pyroptosis in macrophages. The expression of miR-155 was detected by real-time PCR and inhibited using RNAi. Suppressor of cytokine signaling (SOCS) 1, cleaved GSDMD, caspase (CAS)-1, caspase-11, apoptosis-associated speck-like protein (ASC), and NOD-like receptor protein 3 (NLRP3) were detected by Western blotting, and IL-1β and IL-18 were detected by ELISA.

Results: The rate of pyroptosis in macrophages and the expression of miR-155 increased upon stimulation with P. gingivalis and pyroptosis rate decreased when miR-155 was silenced. GSDMD-NT, CAS-11, CAS-1, ASC, NLRP3, IL-1β, and IL-18 levels increased, but SOCS1 decreased in U937 cells after stimulated with P. gingivalis. These changes were weakened in P. gingivalis-stimulated U937 macrophages transfected with lentiviruses carrying miR-155 shRNA compared to those transfected with non-targeting control sequence. However, there was no significant difference in ASC expression between P. gingivalis-stimulated shCont and shMiR-155 cells.

Conclusions: Porphyromonas gingivalis promotes pyroptosis in macrophages during early infection. miR-155 is involved in this process through regulating the NLRP3 inflammasome.

Keywords: Porphyromonas gingivalis; NLRP3; macrophages; miR-155; pyroptosis.