榄仁树提取物及其馏分的抗菌、抗生物膜和细胞毒性活性。
Antibacterial, antibiofilm and cytotoxic activities of Terminalia fagifolia Mart. extract and fractions.
作者信息
de Araujo Alyne Rodrigues, Quelemes Patrick Veras, Perfeito Márcia Luana Gomes, de Lima Luíza Ianny, Sá Melka Coêlho, Nunes Paulo Humberto Moreira, Joanitti Graziella Anselmo, Eaton Peter, Soares Maria José Dos Santos, de Souza de Almeida Leite José Roberto
机构信息
Center for Research on Medicinal Plants, Federal University of Piauí, Piauí, Brazil.
Biodiversity and Biotechnology Research Center, Biotec, Federal University of Piauí, Parnaíba, Piauí, Brazil.
出版信息
Ann Clin Microbiol Antimicrob. 2015 Apr 19;14:25. doi: 10.1186/s12941-015-0084-2.
BACKGROUND
The methicillin resistance of bacteria from the genus Staphylococcus and its ability to form biofilms are important factors in pathogenesis of these microorganisms. Thus, the search for new antimicrobials agents, especially from plants, has been intensified. In this context, Terminalia species have been the subject of research for many pharmacological activities. In this study we evaluated the antibacterial, antibiofilm and cytotoxic activities of the ethanol extract (EtE) from Terminalia fagifolia stem bark as well as that of three fractions of the extract (AqF, HaF and WSF).
METHODS
We determined the minimum inhibitory concentration (MIC) by microdilution in 96-well plates, where the strains were exposed to serial dilutions of the ethanol extract and fractions, ranging from 12.5 to 400 μg/mL. We then determined the minimum bactericidal concentration (MBC), seeding the inoculum (10 μL) with concentrations equal to or greater than the MIC in Mueller-Hinton agar. To test the antibiofilm activity biofilm formation was induced in the presence of concentrations equivalent to 1/2, 1/4 and 1/8 of the MIC extract or fraction tested. In addition, the effect of the EtE and the fractions on cell viability was tested by the MTT assay on human MCF-7 breast cancer and mouse fibroblast NIH/3T3. To obtain high-resolution images of the effect of the aqueous fraction on the bacterial morphology, atomic force microscopy (AFM) imaging of treated S. aureus cells was performed.
RESULTS
We observed antibacterial activity of EtE and fractions with MICs ranging from 25-200 μg/mL and MBCs ranging from 200-400 μg/mL. Regarding antibiofilm activity, both the EtE as the AqF, HaF and WSF fractions showed significant inhibition of the biofilm formation, with inhibition of biofilms formation of over 80% for some strains. The EtE and fractions showed a moderate cytotoxicity in cell line NIH/3T3 viability and potential antitumoral activity on human breast cancer cell line MCF-7. The microscopic images obtained revealed morphological changes to the S. aureus ATCC 29213 surface caused by AqF, as well as significant size alterations.
CONCLUSIONS
The results show potential antibacterial, antibiofilm and antitumoral activities of the ethanol extract and fractions of T. fagifolia.
背景
葡萄球菌属细菌的耐甲氧西林特性及其形成生物膜的能力是这些微生物致病的重要因素。因此，对新型抗菌剂的寻找，尤其是来自植物的抗菌剂，已得到加强。在此背景下，榄仁属植物已成为许多药理活性研究的对象。在本研究中，我们评估了榄仁树茎皮乙醇提取物（EtE）以及该提取物的三个馏分（水相馏分AqF、己烷相馏分HaF和水饱和正丁醇相馏分WSF）的抗菌、抗生物膜和细胞毒性活性。
方法
我们通过在96孔板中进行微量稀释来测定最低抑菌浓度（MIC），将菌株暴露于乙醇提取物和馏分的系列稀释液中，浓度范围为12.5至400μg/mL。然后测定最低杀菌浓度（MBC），将等于或大于MIC浓度的接种物（10μL）接种于 Mueller-Hinton琼脂中。为了测试抗生物膜活性，在相当于所测试提取物或馏分MIC的1/2、1/4和1/8浓度存在下诱导生物膜形成。此外，通过MTT法在人MCF-7乳腺癌细胞和小鼠成纤维细胞NIH/3T3上测试了EtE和馏分对细胞活力的影响。为了获得水相馏分对细菌形态影响的高分辨率图像，对经处理的金黄色葡萄球菌细胞进行了原子力显微镜（AFM）成像。
结果
我们观察到EtE和馏分具有抗菌活性，MIC范围为25 - 200μg/mL，MBC范围为200 - 400μg/mL。关于抗生物膜活性，EtE以及AqF、HaF和WSF馏分均显示出对生物膜形成的显著抑制，对某些菌株的生物膜形成抑制率超过80%。EtE和馏分在细胞系NIH/3T3活力方面表现出中等细胞毒性，对人乳腺癌细胞系MCF-7具有潜在的抗肿瘤活性。获得的显微镜图像显示，AqF导致金黄色葡萄球菌ATCC 29213表面出现形态变化以及显著的大小改变。
结论
结果表明榄仁树乙醇提取物及其馏分具有潜在的抗菌、抗生物膜和抗肿瘤活性。
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