Weissella cibaria CMU exerts an anti‑inflammatory effect by inhibiting Aggregatibacter actinomycetemcomitans‑induced NF‑κB activation in macrophages

Abstract

Periodontitis is a chronic inflammatory disease caused by various periodontal pathogens. Weissella cibaria CMU (oraCMU) is a probiotic that promotes oral health. However, its anti-inflammatory effects against periodontal pathogens have not yet been investigated. The present study evaluated the anti-inflammatory effects of live oraCMU against stimulation with the formalin-inactivated periodontal pathogen Aggregatibacter actinomycetemcomitans in RAW 264.7 macrophages. Cell viability was analyzed by the MTS assay in a dose-dependent manner (at multiplicities of infection of 0.1, 1, 10, 100 and 1,000). Nitric oxide (NO) was monitored using the Griess test. The mRNA expression of proinflammatory cytokines such as interleukin (IL)1β and IL6 was assessed by reverse transcription-quantitative PCR. Western blotting was used to examine the effects of oraCMU on the phosphorylation of NF-κB inhibitor α (iκBα) and iκBα kinase (IKK), the nuclear translocation of the NF-κB subunit p65 and the expression of inducible NO synthase (iNOS). Live oraCMU had no cytotoxic effects on RAW 264.7 macrophages. In A. actinomycetemcomitans-stimulated RAW 264.7 macrophages, oraCMU reduced NO production by suppressing iNOS expression and downregulating the mRNA expression of proinflammatory cytokines in a dose-dependent manner. IKK phosphorylation and IκBα degradation were dose-dependently inhibited by oraCMU and the nuclear translocation of p65 via the canonical NF-κB pathway was simultaneously reduced. The results indicated that oraCMU possessed anti-inflammatory activity associated with the inhibition of NF-κB signal activation in response to periodontal pathogens. This suggests that oraCMU is a beneficial anti-inflammatory probiotic that can aid in the maintenance of oral health.

Assessment of safety and in situ antibacterial activity of Weissella cibaria strains isolated from Campos das Vertentes, Minas Gerais, Brazil for their food application

“…It is noteworthy that no cytotoxicity was found for these Weissella strains against human cell lines tested during our study, matching with other studies that underpinned the safety of W. cibaria strains on the RAW 264.7 macrophage cell line and human mouth epithelial cell line KCLB 10017 (Kim et al, 2020;Kang et al, 2011). We also assessed the in ammatory and anti-in ammatory capacity of Weissella strains against the human U937 cells and these strains showed neither in ammatory nor anti-in ammatory responses.…”

Longitudinal Analysis of the Impacts of Urogenital Schistosomiasis on the Gut microbiota of Adolescents in Nigeria

“…Weissella species have been isolated from raw milk, fermented cereals and vegetables 50 . Some species, such as W. cibaria, have been identi ed to possess anticancer, antiin ammatory, anti-fungal, anti-bacterial and immune-boosting properties 51,52 . W. cibaria is a lactic acid bacterium and has been associated with regulating the integrity of the intestinal epithelial barrier, through dampening in ammatory responses that reduce the synthesis of TNFa, IL-6 and Il-8 51,52 .…”

“…Some species, such as W. cibaria, have been identi ed to possess anticancer, antiin ammatory, anti-fungal, anti-bacterial and immune-boosting properties 51,52 . W. cibaria is a lactic acid bacterium and has been associated with regulating the integrity of the intestinal epithelial barrier, through dampening in ammatory responses that reduce the synthesis of TNFa, IL-6 and Il-8 51,52 . The signi cant reduction in the presence of these bacteria in the S. haematobium infected group might point to increased in ammation in the kids.…”

Probiotic Species in the Management of Periodontal Diseases: An Overview

“…In RAW 264.7 macrophages stimulated by formalin-inactivated A. actinomycetemcomitans ATCC 3338, W. cibaria CMU downregulated the expression of inducible NO synthase (iNOS) and the mRNA of IL-1b and IL-6 to reduce NO production. The inhibition of NF-kB inhibitor a (IkBa) kinase (IKK) phosphorylation, IkBa degradation, and the nuclear translocation of p65 were also observed in W. cibaria CMU-treated RAW 264.7 macrophages (Kim et al, 2020b). In animal models, W. cibaria CMU decreased the level of both proinflammatory (TNF-a, IL-1b, and IL-6) and anti-inflammatory (IL-10) cytokines (Kim et al, 2020a).…”