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麦罗塔木属植物叶中的多酚抑制牙龈卟啉单胞菌的体外黏附并对 KB 细胞发挥抗炎细胞保护作用。

Polyphenols from Myrothamnus flabellifolia Welw. inhibit in vitro adhesion of Porphyromonas gingivalis and exert anti-inflammatory cytoprotective effects in KB cells.

机构信息

Institute of Pharmaceutical Biology and Phytochemistry, University of Münster, Münster, Germany.

出版信息

J Clin Periodontol. 2011 May;38(5):457-69. doi: 10.1111/j.1600-051X.2010.01654.x. Epub 2010 Dec 15.

DOI:10.1111/j.1600-051X.2010.01654.x
PMID:21158896
Abstract

AIM

Identification of anti-adhesive plant extracts against cell surface binding of Porphyromonas gingivalis and underlying mechanisms; investigation of potential cytoprotective effects of anti-adhesive extract on KB cells.

MATERIALS AND METHODS

Polyphenol-enriched extract, fully characterized concerning flavan-3-ols and oligomeric proanthocyanidins, from Myrothamnus flabellifolia (MF), traditionally used for periodontitis, was tested for inhibition of P. gingivalis-mediated adhesion to KB cells by flow cytometry, for influence on gingipain activity (protease assay), haemagglutination and by microarray analysis for effects on bacterial transcriptome. The influence of MF on P. gingivalis-induced cytokine gene expression was monitored by RT-PCR and IL-6 titres by ELISA.

RESULTS

MF (100 μg/ml) reduced P. gingivalis adhesion/invasion about 50% by interacting with bacterial OMPs. As shown by RT-PCR, fimbrillin and Arg-gingipain encoding genes were up-regulated by MF. On the protein level, inhibition (70%) of Arg-gingipain activity was observed, while the corresponding Lys-gingipain was hardly influenced. MF also inhibited haemagglutination. While exposure to P. gingivalis resulted in an increased expression of inflammation-related genes in KB cells, pre-treatment of KB cells with MF evoked cytoprotective effects concerning IL-1β, IL-6, IL-8 and TNF-α gene expression as well as IL-6 release rates. Compounds from the plant extract belonging to the class of proanthocyanidins were shown to be responsible for the observed effects and were characterized for their respective structural features.

CONCLUSIONS

While being cytoprotective, MF exerts anti-adhesive effects against P. gingivalis. Thus, MF may be useful for the prevention of P. gingivalis-associated periodontal diseases.

摘要

目的

鉴定具有抗黏附作用的植物提取物对牙龈卟啉单胞菌细胞表面结合的抑制作用及其潜在机制;研究抗黏附提取物对 KB 细胞的潜在细胞保护作用。

材料和方法

从传统用于治疗牙周炎的 Myrothamnus flabellifolia(MF)中提取多酚富集提取物,该提取物富含黄烷-3-醇和低聚原花青素,并用流式细胞术检测其对 KB 细胞中牙龈卟啉单胞菌介导的黏附作用的抑制作用、对牙龈蛋白酶活性(蛋白酶测定)、血凝作用以及通过微阵列分析对细菌转录组的影响。通过 RT-PCR 监测 MF 对牙龈卟啉单胞菌诱导的细胞因子基因表达的影响,通过 ELISA 监测白细胞介素 6(IL-6)浓度。

结果

MF(100μg/ml)通过与细菌外膜蛋白相互作用,减少约 50%的牙龈卟啉单胞菌黏附/侵袭。如 RT-PCR 所示,MF 上调了 fimbrillin 和 Arg-牙龈蛋白酶编码基因。在蛋白质水平上,观察到 Arg-牙龈蛋白酶活性抑制(70%),而相应的 Lys-牙龈蛋白酶几乎不受影响。MF 还抑制了血凝作用。当 KB 细胞暴露于牙龈卟啉单胞菌时,炎症相关基因的表达增加,而 MF 预处理 KB 细胞可减轻与 IL-1β、IL-6、IL-8 和 TNF-α基因表达以及 IL-6 释放率相关的细胞保护作用。来自植物提取物的属于原花青素类的化合物被证明对观察到的作用负责,并对其各自的结构特征进行了表征。

结论

MF 具有细胞保护作用,同时对牙龈卟啉单胞菌具有抗黏附作用。因此,MF 可能有助于预防与牙龈卟啉单胞菌相关的牙周病。

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