两种基因型牙龈卟啉单胞菌对上皮细胞的影响

The influence of Porphyromonas gingivalis with two different fima genotypes on oral epithelial cells

目的比较2种fimA基因型牙龈卟啉单胞菌(Porphyromonas gingivalis,P.g)刺激下口腔上皮细胞白介素-6(Interlukin-6,IL-6)的表达。方法以未受P.g刺激的上皮细胞作为对照组,实验组用P.g ATCC 33277(I型菌毛组)和W83、47A-1(IV型菌毛组)分别与口腔上皮细胞孵育24 h,于1、3、6、24 h收集细胞和培养上清液。逆转录-多聚酶联反应检测KB细胞IL-6 mRNA的表达,酶联免疫反应检测培养上清液中IL-6的变化。结果 1～24h实验组IL-6mRNA的表达均高于对照组(P<0.05),6h IV型菌毛组IL-6mRNA表达高于I型菌毛组(P<0.05),IL-6 mRNA和蛋白水平表达不一致,3～24h I型菌毛组IL-6蛋白水平高于IV型菌毛组和对照组(P<0.05),提示IL-6的表达存在转录后水平的调节。结论 P.g菌毛基因型与上皮细胞表达细胞因子的水平相关,IV型菌毛的调节作用强于I型菌毛,提示P.g致病性与其fimA基因型相关。

Objective To investigate the mechanism of cytokine regulations of oral epithelial cells by challenge of P.g with different fimA genotypes. Methods P.g ATCC 33277（type I）,W83（type IV）,47A-1（type IV） were assessed for their inductions of IL-6 expression in the human oral epithelial cells.KB cells without the stimulation of P.g were used as control group.mRNA expression was determined by reverse transcription polymerase chain reaction（RT-PCR） at different time intervals（1h,3h,6h and 24h） following continuous co-culture of bacteria with KB cell line,and cytokine protein levels in culture supernatant were determined by ELISA. Results From 1h to 24h,IL-6 mRNA expression in experimental group was higher than that in control group（P0.05）;At 6h,IL-6 mRNA expression in group with type IV was higher than that in group with type I（P0.05）.IL-6 mRNA expression was not consistent with its protein level,from 3h to 24h,IL-6 protein level in group with type I was higher that in group with type I and control group（P0.05）. Conclusion fimA genotypes of P.g is related to the effect of cytokine inductions,and the regulation effect of IV genotype is stronger than that of I genotype,which indicates that fimA genotype is associated with pathogenesis of P.ginivalis.