目的比较不同fimA基因型牙龈卟啉单胞菌(Porphyromonas gingivalis,P.gingivalis)刺激下口腔上皮细胞表达细胞因子的水平。方法P.gingivalis ATCC33277(IfimA),W83(ⅣfimA),47A-1(ⅣfimA)分别与KB细胞ATCCCCL17共同孵育6h,在3h和6h收集细胞和培养上清液。逆转录-聚合酶链式反应检测KB细胞IL-8,IL-6,IL-1βmRNA的表达,酶联免疫反应检测培养上清液中IL-8,IL-6,IL-1β的变化。结果3h时,ⅣfimA组IL-6蛋白水平低于IfimA组(P〈0.05),6h时,ⅣfimA组IL-6和IL-8蛋白水平均低于ⅠfimA组(P〈0.05);IL-8,IL-6mRNA和蛋白水平表达不一致,提示存在转录后水平的调节。3h,6h时,IL-1β对P.gingvalis刺激不敏感,mRNA和蛋白表达水平都很低。结论P.gingivalis fimA基因型与上皮细胞表达细胞因子的水平相关,提示P.gingivalis致病性与其fimA基因型相关。
Objective To investigate the regulations of cytokines in oral epithelial cells by P. gingivalis with differentfimA genotypes. Methods P. gingivalis ATCC 33277 ( type I ), W83 ( type Ⅳ ), 47A - 1 ( type Ⅳ ) were added to the culture of human oral epithelial cells for 3h or 6h. The expressions of IL-6, IL- 8 and IL- 1β mRNA were determined by reverse transcription polymerase chain reaction ( RT - PCR). The corresponding protein levels in culture supematant was determined by ELISA. Results. After treated for 3h, IL -6 protein level in group with IV genotype was lower than that of I genotype. After treated for 6h, IL - 8 and IL - 6 protein levels in group with IV genotype both were lower than those of I genotype ( P 〈 0.05). However, the expressions of IL -8 and IL -6 mRNA were not consistent with their protein levels, which indicated post -transcriptional regulations ( P 〈0.05). After treated for 3h or 6h, all the IL - 1β mRNA and protein expressions were low. Conclusion fimA genotypes of P. gingivalis showed the different effects on cytokines induction in oral epithelial cells, suggesting thatfimA genotype may be associated with the pathogenesis of P. ginivalis .