Effect of periodontitis patient's own tissue nucleic acid on expression of NLRP3 mRNA in murine macrophages and its significance / 吉林大学学报(医学版)

ABSTRACT

Objective:To detect the mRNA expression levels of inflammatory-related factors NLRP3, caspase-1, IL-1β, and IL-18in the murine macrophages infected by periodontitis patient's own tissue nucleic acid, and to discuss the effects of periodontitis patient's own tissue nucleic acid on the inflammation-related factors in the macrophages.Methods:The inflammatory periodontal tissue samples were collected during periodontal flap surgery of the chronic periodontitis patients, and the healthy periodontal tissue samples were collected from the patients without any periodontal diseases undergoing crown lengthening surgery.Then the total RNA from gingival tissue was extracted and reversely transcribed into cDNA.The cultured mouse macrophages RAW264.7were divided into control group and experiment group, then the healthy periodontal tissue cDNA and inflammatory periodontal tissue cDNA (the cDNA at a concentration of 1mg·L-1) were added into the RAW264.7cells, respectively.Real-time PCR was used to detect the mRNA expression levels of NLRP3, Caspase-1, IL-1β, and IL-18in the macrophages in various groups at 4, 6and 8hafter incubation.Results:The microscope observation showed that the mouse macrophages RAW264.7grew well with round and polygon shapes, clear cytoplasm, and full cell body.Compared with control group, the expression levels of NLRP3, Caspase-1, IL-1β, and IL-18mRNA in the RAW264.7cells in experiment group at 4, 6, and 8hwere increased significantly (P＜0.05or P＜0.01) , and the expression levels of NLRP3, Caspase-1, IL-1β, and IL-18 mRNA in RAW264.7 cells at 6 hin experiment group were the highest.Conclusion:The periodontitis patient's own tissue nucleic acids can promote the mRNA expressions of inflammation-related factors in the RAW264.7cells, suggesting that the periodontitis patient's own tissue nucleic acid has an immunomodulatory effect on the activation of RAW264.7cells.