目的:研究不同fimA基因型牙龈卟啉单胞菌(Porphyromonas gingivalis,Pg)对人牙龈成纤维细胞表达IL-8的影响,探讨fimA基因型与Pg致病力差异之间的关系。方法:PgATCC 33277(Ⅰ型),WCSP115(Ⅱ型),WCSP1.5(Ⅲ型),W83(Ⅳ型)分别与牙龈成纤维细胞在标准条件下共同孵育,于孵育后1、3、6、12 h收集细胞和上清,应用实时荧光定量RT-PCR和ELISA法分别检测牙龈成纤维细胞IL-8 mRNA和蛋白的动态表达。结果:与对照组比较,各fimA型Pg对牙龈成纤维细胞IL-8mRNA和蛋白的表达均有明显的促进作用(P<0.01);其中ⅡfimA型组IL-8 mRNA和蛋白的表达量明显高于其它fimA型组,不同时间点IL-8 mRNA相对表达量分别为20.42±2.21~103.01±12.50,蛋白分泌水平为(137.46±4.97~323.24±13.74)pg/ml;而ⅢfimA型Pg组的表达水平弱于其它fimA型组,IL-8 mRNA相对表达量为3.61±0.39~12.88±1.56,蛋白分泌水平为(44.83±3.68~189.19±87.34)pg/ml,差异有统计学意义(P〈0.05)。结论:Pg可促进牙龈成纤维细胞IL-8mRNA和蛋白的表达,且各fimA型Pg的促进作用有所差异。提示:fimA基因型的不同可能是pg的致病力差异的基因基础。
Objective:To investigate the mechanism of IL-8 expressions in human gingival fibroblasts(HGFs) regulated by Porphyromonas gingivalis(Pg) with different fimA genotypes.Methods:Pg ATCC 33277(typeⅠ),WCSP115(type Ⅱ),WCSP1.5(type Ⅲ),W83(type Ⅳ) were assessed for their inductions of IL-8 expression in HGFs.IL-8 mRNA levels of HGFs were determined by Real-time RT-PCR and protein levels in culture supernatant were determined by ELISA at different time intervals(1,3,6 and 12 h) following continuous co-culture of bacteria with HGFs. Results: The IL-8 mRNA and protein expressions of HGFs co-cultured with Pgwere significantly increased compared with the negative control group (P 〈 0. 01 ). The group of type Ⅱ showed greater up-regulation than otherfimA genotypes on the mRNA and protein expressions of IL-8, mRNA 20.42 ±2.21 ~ 103.01 ± 12.50 and protein ( 137.46 ±4.97 ~ 323.24 ±13.74) pg/ml for different time periods, while the group of type 11I was weaker than otherfimA genotypes, the level of IL-8 mRNA 3.61 ± 0.39 ~12.88 ± 1.56 and protein (44.83 ± 3.68 ~ 189.19 ±87.34 ) pg/ml (P 〈 0.05 ). Conclusion: Pg could induce HGF to over-express IL-8, fimA genotypes of Pg may be related with this pathogenicity, which might indicatefimA genotype is associated with pathogenesis of Pg.