Outer membrane vesicles of Porphyromonas gingivalis trigger NLRP3 inflammasome and induce neuroinflammation, tau phosphorylation, and memory dysfunction in mice

Abstract

BackgroundPorphyromonas gingivalis (Pg), the keystone pathogen in chronic periodontitis, is reported to initiate Alzheimer’s disease pathologies in preclinical studies. However, the specific mechanisms and signaling pathways acting on the brain still need to be further explored. Outer membrane vesicles are derived from Gram-negative bacteria and contain many virulence factors of bacteria. We hypothesized that outer membrane vesicles are an important weapon of Porphyromonas gingivalis to initiate Alzheimer’s disease pathologies.MethodsThe outer membrane vesicles of Porphyromonas gingivalis (Pg OMVs, 4 mg/kg) or saline were delivered to 14-month-old mice by oral gavage every other day for eight weeks. Behavioral alterations were assessed by the open field test, Morris water maze, and Y-maze test. Blood–brain barrier permeability, neuroinflammation, tau phosphorylation, and NLRP3 inflammasome-related protein were analyzed.ResultsPg OMVs impaired memory and learning ability of mice and decreased tight junction–related gene expression ZO-1, occludin, claudin-5, and occludin protein expression in the hippocampus. Pg OMVs could be detected in the hippocampus and cortex three days after oral gavage. Furthermore, Pg OMVs activated both astrocytes and microglia and elevated IL-1β, tau phosphorylation on the Thr231 site, and NLRP3 inflammasome–related protein expression in the hippocampus. In in vitro studies, Pg OMV (5 µg/ml) stimulation increased the mRNA and immunofluorescence of NLRP3 in BV2 microglia, which were significantly inhibited by the NLRP3 inhibitor MCC950. In contrast, the tau phosphorylation in N2a neurons was enhanced after treatment with conditioned media from Pg OMV-stimulated microglia, which was attenuated after pretreatment with MCC950.ConclusionsThese results indicate that Pg OMVs prompt memory dysfunction, neuroinflammation, and tau phosphorylation and trigger NLRP3 inflammasome in the brain of middle-aged mice. We propose that Pg OMVs play an important role in activating neuroinflammation in the AD-like pathology triggered by Porphyromonas gingivalis, and NLRP3 inflammasome activation is a possible mechanism.

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“…Neuroinflammation is existent throughout the course of AD. Among the innate immune cells in the central nervous system (CNS), microglia are the primary participants in neuroinflammation [ 41 ]. Microglia possess similar functions to peripheral myeloid cells including chemotaxis, phagocytosis, and cytokine expression.…”

The role of periodontitis‐associated bacteria in Alzheimer's disease: A narrative review

“…The outer membrane vesicles of P. gingivalis lower cytoplasmic proteins and occlude gene expression. This helps compounds such as outer membrane vesicles enter the brain and disrupt the BBB [101]. P. gingivalis can directly enter the brain through a damaged BBB, cause neuro-inflammation and release inflammatory cytokines [102].…”

The role of extracellular vesicles in periodontitis: pathogenesis, diagnosis, and therapy

“…The Gram-negative bacteria that are closely related to the progression of periodontitis, including Porphyromonas gingivalis (P. gingivalis), Treponema denticola (T. denticola), Tannerella forsythia (T. forsythia), Actinomyces reticulata (A. reticulata), Fusobacterium nucleatum (F. nucleatum) and Prevotella intermedia (P. intermedia) have been isolated from the periodontal pocket ( 94 – 96 ). P. gingivalis is the primary pathogen responsible for chronic periodontitis ( 97 , 98 ). It forms the “red complex” along with T. forsythia and T. denticola , which are accessory pathogens with complementary or supplementary functions ( 99 , 100 ).…”