Inhibitory effects of macrocarpals on the biological activity of Porphyromonas gingivalis and other periodontopathic bacteria
Inhibitory effects of macrocarpals on the biological activity of Porphyromonas gingivalis and other periodontopathic bacteria
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DOI：
10.1111/j.1399-302x.2006.00269.x
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发表时间：
2006-06-01
影响因子：
--
通讯作者：
Shizukuishi, S
中科院分区：
文献类型：
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作者：
Nagata, H;Inagaki, Y;Shizukuishi, S
背景/目的：巨腕骨是桉树叶中所含的间苯三酚衍生物，对包括口腔细菌在内的多种细菌具有抗菌活性。本研究考察了大腕骨A、B和C对牙周病细菌，特别是牙龈卟啉单胞菌的影响。方法：从桉叶60%乙醇提取物中纯化大腕骨A、B和C。为了研究抗菌活性，将代表性牙周病细菌培养在有或没有不同数量大腕骨的培养基中；随后，测量了660 nm处的光密度。采用荧光分光光度法和十二烷基硫酸钠-聚丙烯酰胺凝胶电泳法测定牙龈假单胞菌精氨酸和赖氨酸特异性蛋白酶的大腕抑制作用。用h -3标记的牙龈卟啉单胞菌研究了大腕骨对牙龈卟啉单胞菌与唾液包被羟基磷灰石珠结合的影响。结果：大腕骨对牙龈假单胞菌生长的抑制作用强于对中间普雷沃菌、黑变普雷沃菌和密螺旋体的抑制作用，而对放线菌和核梭杆菌的抑制作用更强。大腕果对牙龈假单胞菌精氨酸和赖氨酸特异性蛋白酶的抑制呈剂量依赖性。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析，证实了大腕膜的酶抑制作用，大腕膜抑制了牙龈卟啉卟啉蛋白酶对血红蛋白的降解。牙龈卟啉卟啉与唾液包被羟基磷灰石珠的结合也被大腕骨强烈减弱。结论：大腕骨A、B和C对牙周病细菌具有抗菌活性。在被试细菌中，牙龈假单胞菌对大腕骨最敏感；此外，其胰蛋白酶样蛋白酶活性和与唾液包被羟基磷灰石珠的结合受到大腕骨的抑制。这些结果表明，桉树叶提取物可能是有效的预防牙周病。
Background/aims: Macrocarpals, which are phloroglucinol derivatives contained in eucalyptus leaves, exhibit antimicrobial activity against a variety of bacteria including oral bacteria. This study examined effects of macrocarpals A, B, and C on periodontopathic bacteria, especially Porphyromonas gingivalis.Methods:Macrocarpals A, B, and C were purified from a 60% ethanol-extract of Eucalyptus globules leaves. To investigate antibacterial activity, representative periodontopathic bacteria were cultured in media with or without various amounts of macrocarpals; subsequently, the optical density at 660 nm was measured. Macrocarpal inhibition of P. gingivalis Arg- and Lys-specific proteinases was assessed by spectrofluorophotometric assay and sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis. The effect of macrocarpals on P. gingivalis binding to saliva-coated hydroxyapatite beads was examined with H-3-labeled P. gingivalis.Results: Growth of P. gingivalis was inhibited more strongly than growth of Prevotella intermedia or Prevotella nigrescens and Treponema denticola by macrocarpals, however, Actinobacillus actinomycetemcomitans and Fusobacterium nucleatum were much more resistant. Macrocarpals inhibited P. gingivalis Arg- and Lys-specific proteinases in a dose-dependent manner. The enzyme-inhibitory effect of macrocarpals was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis in which hemoglobin degradation by P. gingivalis proteinase was inhibited by macrocarpals. P. gingivalis binding to saliva-coated hydroxyapatite beads was also strongly attenuated by macrocarpals.Conclusions: Macrocarpals A, B and C demonstrated antibacterial activity against periodontopathic bacteria. Among tested bacteria, P. gingivalis displayed the greatest sensitivity to macrocarpals; additionally, its trypsin-like proteinase activity and binding to saliva-coated hydroxyapatite beads were inhibited by macrocarpals. These results indicate that eucalyptus leaf extracts may be useful as a potent preventative of periodontal disease.