目的比较不同fimA基因型牙龈卟啉单胞菌(P.gingivalis)刺激下人外周血中性粒细胞(PMNs)产生基质金属蛋白酶8、9(MMP-8、MMP-9)的差异。方法采用密度梯度离心分离法分离纯化培养PMNs,P.gingivalis ATCC33277(ⅠfimA)、WCSP115(ⅡfimA)、WCSP1.5(Ⅲ fimA)、W83(ⅣfimA)、WCSP559(ⅣfimA)菌悬液与新鲜分离的PMNs悬液共孵育2h,于5min、30min、1h、2h收集上清液,用ELISA方法检测MMP-8、MMP-9的表达。结果不同fimA基因型P.gingivalis刺激PMNs产生MMP-8、MMP-9的能力都显著高于未受刺激组;ⅡfimA、ⅣfimA(W83)型P.gingivalis刺激PMNs分泌MMP-8在速度及量上均高于ⅢfimA、ⅣfimA(WCSP559)型Rgingivalis:ⅠfimA、ⅡfimA、ⅣfimA(W83)型P.gingivalis刺激PMNs分泌MMP-9能力高于ⅢfimA、ⅣfimA(WCSP559)型P.gingivalis。结论ⅡfimA、ⅣfimA型P.gingivalis的致病能力相对较强,提示P.gingivalis的毒力和致病性与fimA基因型存在相关性。
Objective To investigate the pathogenicity of matrix metalloproteinase 8, 9 (MMP-8, MMP-9) regulations of polymorphonuclear leukocytes (PMNs) by challenge of Porphyromonas gingivalis (P.gingivalis) with different fimA genotypes. Methods The studies mainly adopt the isopycnic sedimentation separation to separate the PMNs from human peripheral blood. P.gingivalis ATCC 33277 (type Ⅰ ), WCSP 115 (type Ⅱ ), WCSP 1.5 (typeⅢ), W83 (typelV), WCSP 559 (type Ⅳ) were assessed for their inductions of MMP-8, MMP-9 expression in PMNs. MMP-8, MMP-9 protein levels in culture supernatant were determined by ELISA at different time intervals (5 min, 30 min, 1 h, 2 h) following continuous co-culture of bacteria with PMNs. Results MMP-8 and MMP-9 protein levels produced by PMNs co-culture with the Ⅰ fimA--ⅣfimA P.gingivalis were significantly stronger than unsimulated group. The velocity and quantity of MMP-8 produced by PMNs co-culture with the ⅡfimA P.gingivalis and ⅣfimA P.gingivalis were more than ⅢfimA, ⅣfimA P.gingivalis. The MMP-9 protein levels produced by PMNs co-culture with the Ⅰ fimA, Ⅱ fimA, ⅣfimA P.gingivalis was significantly stronger than ⅢfimA and ⅣfimA P.gingivalis. Conclusion Ⅱ fimA and Ⅳ fimA P.gingivalis have stronger pathogenicity relatively, which indicate that fimA genotype is associated with pathogenesis of P.gingivalis.