Pasteurella multocida Toxin Aggravates Ligatured-Induced Periodontal Bone Loss and Inflammation via NOD-Like Receptor Protein 3 Inflammasome

Figure 3

Altered oral microbiota between WT and Nlrp3^-/- mice and PMT induces transcription and expression of NLRP3 and inflammatory reaction in vitro. (a)16S rRNA gene sequencing analysis show the general discrete clustering pattern of bacterial taxa between WT and Nlrp3^-/- mice (/group). (b) qRT-PCR validation of Pasteurella multocida in WT and Nlrp3^-/- mice (/group). (c) mRNA expression of IL-1β, IL-6, and NLRP3 after the stimulation of PMT at a series of time by qRT-PCR (/group). GAPDH was used for normalization relative to control groups. (d) Protein expression of NLRP3 and the internal control β-actin after the stimulation of PMT at a series of time by western blot. The histogram shows the quantification of band intensities. β-Actin was used for normalization relative to the control groups. (e) Protein expression of pp65, p65, IκBα, and the internal control β-actin after stimulation of PMT with or without pretreatment of NF-κB inhibitor Bay 11-7082 by western blot. The histogram shows the quantification of band intensities. β-Actin was used for normalization relative to the control groups. Values are presented as (, , and ). Abbreviations: GAPDH = glyceraldehyde-3-phosphate dehydrogenase; IL = interleukin; mRNA = messenger RNA; NLRP3 = NOD-like receptor family pyrin domain-containing 3; NF-κB = nuclear factor kappa light chain enhancer of B cells; PMT = Pasteurella multocida toxin; qRT-PCR = quantitative reverse-transcription polymerase chain reaction.

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