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目的探讨慢性牙周炎患者牙龈卟啉单胞菌(porphyromonas gingivalis,Pg)感染与NLRP3、L-1β及IL-18在牙周膜细胞中的表达。方法选取诊治慢性牙周炎需进行拔牙治疗的患者50例及50例牙周健康人群,实时荧光PCR技术检测龈下菌斑Pg,慢性牙周炎患者拔牙后取得牙周膜组织进行牙周膜细胞培养,取部分细胞用Pg标准株感染,部分不做感染处理,采用蛋白印迹法检测NLRPS蛋白表达,以免疫酶联吸附法检测IL-1β及IL-18表达。结果慢性牙周炎患者Pg阳性率为84.00%,高于牙周健康者的22.00%,探诊深度5、6、7 mm慢性牙周炎患者Pg浓度分别为4.38±0.81、4.79±0.76、5.47±0.63,高于牙周健康者0.38±0.12、0.46±0.15、0.72±0.20(P<0.05);Pg感染牙周膜细胞中NLRP3及IL-1β、IL-18表达量分别为(1.25±0.12)、(1 820.42±425.57)pg/ml、(311.55±45.70)pg/ml,明显高于未人工Pg感染牙周膜细胞(0.74±0.08)、(1 138.65±350.76)pg/ml、(124.70±42.54)pg/ml(P<0.05)。结论慢性牙周炎患者Pg感染率及感染菌量均较高,Pg感染可引起牙周膜细胞中NLRP3、IL-1β、IL-18表达的上调,这可能是Pg引起慢性牙周炎发生的原因之一。
Objective To investigate the expression of porcine pathogens Porphyromonas gingivalis (Pg), NLRP3, L-1β and IL-18 in periodontal ligament cells in patients with chronic periodontitis. Methods 50 patients with chronic periodontitis requiring tooth extraction and 50 healthy people with periodontal disease were selected. The subgingival plaque Pg was detected by real-time fluorescence PCR and the periodontal ligament tissue was harvested after periodontal extraction for periodontal ligament cell culture , Some cells were infected with Pg standard strain, some were not infected, NLRPS protein expression was detected by Western blotting, and the expression of IL-1β and IL-18 was detected by enzyme-linked immunosorbent assay. Results The positive rate of Pg in patients with chronic periodontitis was 84.00%, which was higher than that of periodontal healthy persons (22.00%). The Pg concentrations of patients with chronic periodontitis with probing depth of 5, 6 and 7 mm were 4.38 ± 0.81, 4.79 ± 0.76 and 5.47 ± 0.63, higher than that of periodontal healthy persons (0.38 ± 0.12,0.46 ± 0.15,0.72 ± 0.20, P <0.05). The expression of NLRP3, IL-1β and IL-18 in Pg-infected periodontal ligament cells were 1.25 ± 0.12 ) (1 820.42 ± 425.57) pg / ml and (311.55 ± 45.70) pg / ml, respectively, which were significantly higher than those of the non-artificial Pg infected periodontal ligament cells (0.74 ± 0.08), (1 138.65 ± 350.76) pg / ml, ± 42.54) pg / ml (P <0.05). Conclusion Pg infection and bacterial infection in patients with chronic periodontitis are high. Pg infection may induce the up-regulation of NLRP3, IL-1β and IL-18 in periodontal ligament cells, which may be caused by Pg-induced chronic periodontitis one of the reasons.