Inhibitory effects of azithromycin on the adherence ability of Porphyromonas gingivalis
Inhibitory effects of azithromycin on the adherence ability of Porphyromonas gingivalis
复制标题
DOI：
10.1002/jper.18-0559
复制
发表时间：
2019-08-01
影响因子：
4.3
通讯作者：
Minabe, Masato
中科院分区：
文献类型：
--
作者：
Kan, Powen;Sasaki, Haruka;Minabe, Masato
背景牙龈卟啉单胞菌是牙周病的主要病原菌，检出率很高。菌毛和血凝素是由牙龈假单胞菌表达的，它们在牙周组织的粘附和生物膜的形成中起重要作用。本研究的目的是研究亚最低抑制浓度（亚mic）阿奇霉素对牙龈假单胞菌粘附的影响，重点是对菌毛表达和血凝素活性的抑制。方法将牙龈卟啉单胞菌ATCC 33277与阿奇霉素亚mic在37℃下轻度摇培养18 h。收集细菌细胞，用磷酸盐缓冲盐水（PBS）洗涤两次，用12%十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）和western blotting分析蛋白质。黏附试验和血凝素活性试验用相同的培养。结果SDS-PAGE结果显示，阿奇霉素亚mic抑制41-kDa毛膜蛋白表达和血凝素活性。western blot和透射电镜观察证实，在0.4 μ g/mL、0.2 μ g/mL和0.1 μ g/mL的阿奇霉素中，41-kDa毛毡蛋白表达和长毛毡消失。阿奇霉素亚mic可降低牙龈假单胞菌对人牙龈上皮细胞的粘附程度。结论阿奇霉素的亚mic可能通过抑制菌毛的产生和血凝素活性来降低牙龈假单胞菌对宿主细胞的粘附。因此，阿奇霉素可作为一种生物膜治疗牙龈卟啉卟啉菌引起的牙周病。
Background Porphyromonas gingivalis is a major pathogen and has a high detection rate in periodontal disease. Fimbriae and hemagglutinin are expressed by P. gingivalis, and these play an important role in the adherence of the bacteria to periodontal tissue and biofilm formation. The aim of this study was to investigate the effects of sub-minimal inhibitory concentrations (sub-MICs) of azithromycin on the adherence of P. gingivalis, focusing on the inhibition of fimbriae expression and hemagglutinin activity. Methods P. gingivalis ATCC 33277 were incubated anaerobically with sub-MICs of azithromycin at 37 degrees C by gentle shaking for 18 hours. The bacterial cells were harvested, washed twice with phosphate-buffered saline (PBS), and the proteins analyzed by 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and western blotting. Adherence assay and hemagglutinin activity tests were done with the same culture. Results The results of SDS-PAGE indicated that the sub-MICs of azithromycin inhibited 41-kDa fimbrial protein expression and hemagglutinin activities. The disappearance of 41-kDa fimbrial protein expression and long fimbriae in 0.4 mu g/mL, 0.2 mu g/mL, and 0.1 mu g/mL of azithromycin was confirmed by western blotting and transmission electron microscopy. The adherence of P. gingivalis to human gingival epithelial cells was reduced by sub-MICs of azithromycin compared with the adherence levels without antibiotic. Conclusions These results suggest that sub-MICs of azithromycin may reduce the adherence of P. gingivalis to host cells, by inhibiting production of fimbriae and hemagglutinin activities. Therefore, azithromycin can be used as a biofilm treatment of periodontal disease caused by P. gingivalis.