Nagata H, Inagaki Y, Yamamoto Y, Maeda K, Kataoka K, Osawa K, Shizukuishi S
Department of Preventive Dentistry, Osaka University Graduate School of Dentistry, Osaka, Japan.
Oral Microbiol Immunol. 2006 Jun;21(3):159-63. doi: 10.1111/j.1399-302X.2006.00269.x.
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.
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 (3)H-labeled P. gingivalis.
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.
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.
背景/目的：大叶桉醇是桉树叶中含有的间苯三酚衍生物，对包括口腔细菌在内的多种细菌具有抗菌活性。本研究考察了大叶桉醇A、B和C对牙周病原菌，尤其是牙龈卟啉单胞菌的作用。
从蓝桉叶60%乙醇提取物中纯化出大叶桉醇A、B和C。为研究抗菌活性，将代表性牙周病原菌在添加或不添加不同量大叶桉醇的培养基中培养；随后，测量660nm处的光密度。通过荧光分光光度法和十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析评估大叶桉醇对牙龈卟啉单胞菌精氨酸和赖氨酸特异性蛋白酶的抑制作用。用³H标记的牙龈卟啉单胞菌考察大叶桉醇对其与唾液包被羟基磷灰石珠结合的影响。
大叶桉醇对牙龈卟啉单胞菌生长的抑制作用比对中间普氏菌、变黑普氏菌和具核梭杆菌更强，然而，伴放线聚集杆菌和具核梭杆菌的耐药性更强。大叶桉醇以剂量依赖方式抑制牙龈卟啉单胞菌精氨酸和赖氨酸特异性蛋白酶。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析证实了大叶桉醇的酶抑制作用，其中大叶桉醇抑制了牙龈卟啉单胞菌蛋白酶对血红蛋白的降解。大叶桉醇也显著减弱了牙龈卟啉单胞菌与唾液包被羟基磷灰石珠的结合。
大叶桉醇A、B和C对牙周病原菌具有抗菌活性。在所测试的细菌中，牙龈卟啉单胞菌对大叶桉醇的敏感性最高；此外，其类胰蛋白酶活性以及与唾液包被羟基磷灰石珠的结合受到大叶桉醇的抑制。这些结果表明桉树叶提取物可能作为预防牙周疾病的有效药物。
