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几种苏丹药用植物对牙龈卟啉单胞菌和基质金属蛋白酶-9的抑制活性以及从哈特曼风车子(Combretum hartmannianum (Schweinf))树皮中分离生物活性化合物

Inhibitory activities of selected Sudanese medicinal plants on Porphyromonas gingivalis and matrix metalloproteinase-9 and isolation of bioactive compounds from Combretum hartmannianum (Schweinf) bark.

作者信息

Mohieldin Ebtihal Abdalla M, Muddathir Ali Mahmoud, Mitsunaga Tohru

机构信息

Faculty of Pharmacy, University of Science and Technology, Omdurman, Sudan.

Department of Applied Biological Science, Faculty of Applied Biological Science, Gifu University, 1-1 Yanagido, Gifu, 501-1193, Japan.

出版信息

BMC Complement Altern Med. 2017 Apr 20;17(1):224. doi: 10.1186/s12906-017-1735-y.

Abstract

BACKGROUND

Periodontal diseases are one of the major health problems and among the most important preventable global infectious diseases. Porphyromonas gingivalis is an anaerobic Gram-negative bacterium which has been strongly implicated in the etiology of periodontitis. Additionally, matrix metalloproteinases-9 (MMP-9) is an important factor contributing to periodontal tissue destruction by a variety of mechanisms. The purpose of this study was to evaluate the selected Sudanese medicinal plants against P. gingivalis bacteria and their inhibitory activities on MMP-9.

METHODS

Sixty two methanolic and 50% ethanolic extracts from 24 plants species were tested for antibacterial activity against P. gingivalis using microplate dilution assay method to determine the minimum inhibitory concentration (MIC). The inhibitory activity of seven methanol extracts selected from the 62 extracts against MMP-9 was determined by Colorimetric Drug Discovery Kit. In search of bioactive lead compounds, Combretum hartmannianum bark which was found to be within the most active plant extracts was subjected to various chromatographic (medium pressure liquid chromatography, column chromatography on a Sephadex LH-20, preparative high performance liquid chromatography) and spectroscopic methods (liquid chromatography-mass spectrometry, Nuclear Magnetic Resonance (NMR)) to isolate and characterize flavogalonic acid dilactone and terchebulin as bioactive compounds.

RESULTS

About 80% of the crude extracts provided a MIC value ≤4 mg/ml against bacteria. The extracts which revealed the highest potency were: methanolic extracts of Terminalia laxiflora (wood; MIC = 0.25 mg/ml) followed by Acacia totrtilis (bark), Ambrosia maritima (aerial part), Argemone mexicana (seed), C. hartmannianum (bark), Terminalia brownii (wood) and 50% ethanolic extract of T. brownii (bark) with MIC values of 0.5 mg/ml. T. laxiflora (wood) and C. hartmannianum (bark) which belong to combretaceae family showed an inhibitory activity over 50% at the concentration of 10 μg/ml against MMP-9. Additionally, MMP-9 was significantly inhibited by terchebulin with IC value of 6.7 μM.

CONCLUSIONS

To the best of our knowledge, flavogalonic acid dilactone and terchebulin were isolated from C. hartmannianium bark for the first time in this study. Because of terchebulin and some crude extracts acting on P. gingivalis bacteria and MMP-9 enzyme that would make them promising natural preference for preventing and treating periodontal diseases.

摘要

背景

牙周疾病是主要的健康问题之一,也是全球最重要的可预防传染病之一。牙龈卟啉单胞菌是一种厌氧革兰氏阴性菌,与牙周炎的病因密切相关。此外,基质金属蛋白酶-9(MMP-9)是通过多种机制导致牙周组织破坏的一个重要因素。本研究的目的是评估所选苏丹药用植物对牙龈卟啉单胞菌的抗菌活性及其对MMP-9的抑制活性。

方法

采用微孔板稀释法测定24种植物的62种甲醇提取物和50%乙醇提取物对牙龈卟啉单胞菌的抗菌活性,以确定最低抑菌浓度(MIC)。使用比色药物发现试剂盒测定从62种提取物中选出的7种甲醇提取物对MMP-9的抑制活性。为寻找生物活性先导化合物,对活性最强的植物提取物之一的哈氏风车子树皮进行各种色谱法(中压液相色谱法、葡聚糖凝胶LH-20柱色谱法、制备型高效液相色谱法)和光谱法(液相色谱-质谱联用、核磁共振(NMR)),以分离和鉴定黄酮加洛内酯和特切布林为生物活性化合物。

结果

约80%的粗提物对细菌的MIC值≤4mg/ml。效力最高的提取物为:疏花榄仁树(木材)的甲醇提取物(MIC = 0.25mg/ml),其次是阿拉伯金合欢(树皮)、滨海豚草(地上部分)、墨西哥蓟罂粟(种子)、哈氏风车子(树皮)、布朗榄仁树(木材)以及布朗榄仁树(树皮)的50%乙醇提取物,MIC值为0.5mg/ml。属于使君子科的疏花榄仁树(木材)和哈氏风车子(树皮)在浓度为10μg/ml时对MMP-9的抑制活性超过50%。此外,特切布林对MMP-9有显著抑制作用,IC值为6.7μM。

结论

据我们所知,本研究首次从哈氏风车子树皮中分离出黄酮加洛内酯和特切布林。由于特切布林和一些粗提物对牙龈卟啉单胞菌和MMP-9酶有作用,这将使它们成为预防和治疗牙周疾病有前景的天然选择。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3288/5399347/b71983d966b7/12906_2017_1735_Fig1_HTML.jpg

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