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多花萝芙木中吲哚生物碱的抗菌活性

Antibacterial activity of indole alkaloids from Aspidosperma ramiflorum.

作者信息

Tanaka J C A, Silva C C da, Oliveira A J B de, Nakamura C V, Dias Filho B P

机构信息

Departamento de Química, Universidade Estadual de Maringá, Maringá, PR, Brazil.

出版信息

Braz J Med Biol Res. 2006 Mar;39(3):387-91. doi: 10.1590/s0100-879x2006000300009. Epub 2006 Feb 22.

Abstract

We evaluated the antibacterial activities of the crude methanol extract, fractions (I-V) obtained after acid-base extraction and pure compounds from the stem bark of Aspidosperma ramiflorum. The minimum inhibitory concentration (MIC) was determined by the microdilution technique in Mueller-Hinton broth. Inoculates were prepared in this medium from 24-h broth cultures of bacteria (10(7) CFU/mL). Microtiter plates were incubated at 37 masculineC and the MICs were recorded after 24 h of incubation. Two susceptibility endpoints were recorded for each isolate. The crude methanol extract presented moderate activity against the Gram-positive bacteria B. subtilis (MIC = 250 microg/mL) and S. aureus (MIC = 500 microg/mL), and was inactive against the Gram-negative bacteria E. coli and P. aeruginosa (MIC > 1000 microg/mL). Fractions I and II were inactive against standard strains at concentrations of < or =1000 microg/mL and fraction III displayed moderate antibacterial activity against B. subtilis (MIC = 500 microg/mL) and S. aureus (MIC = 250 microg/mL). Fraction IV showed high activity against B. subtilis and S. aureus (MIC = 15.6 microg/mL) and moderate activity against E. coli and P. aeruginosa (MIC = 250 microg/mL). Fraction V presented high activity against B. subtilis (MIC = 15.6 microg/mL) and S. aureus (MIC = 31.3 microg/mL) and was inactive against Gram-negative bacteria (MIC > 1000 microg/mL). Fractions III, IV and V were then submitted to bioassay-guided fractionation by silica gel column chromatography, yielding individual purified ramiflorines A and B. Both ramiflorines showed significant activity against S. aureus (MIC = 25 microg/mL) and E. faecalis (MIC = 50 microg/mL), with EC50 of 8 and 2.5 microg/mL for ramiflorines A and B, respectively, against S. aureus. These results are promising, showing that these compounds are biologically active against Gram-positive bacteria.

摘要

我们评估了粗甲醇提取物、酸碱萃取后得到的馏分(I - V)以及来自多花萝芙木茎皮的纯化合物的抗菌活性。最低抑菌浓度(MIC)通过在 Mueller - Hinton 肉汤中的微量稀释技术测定。接种物由细菌的 24 小时肉汤培养物(10⁷ CFU/mL)在该培养基中制备。微量滴定板在 37℃孵育,孵育 24 小时后记录 MIC。每个分离株记录两个敏感性终点。粗甲醇提取物对革兰氏阳性菌枯草芽孢杆菌(MIC = 250 μg/mL)和金黄色葡萄球菌(MIC = 500 μg/mL)呈现中等活性,对革兰氏阴性菌大肠杆菌和铜绿假单胞菌无活性(MIC > 1000 μg/mL)。馏分 I 和 II 在浓度≤1000 μg/mL 时对标准菌株无活性,馏分 III 对枯草芽孢杆菌(MIC = 500 μg/mL)和金黄色葡萄球菌(MIC = 250 μg/mL)呈现中等抗菌活性。馏分 IV 对枯草芽孢杆菌和金黄色葡萄球菌显示高活性(MIC = 15.6 μg/mL),对大肠杆菌和铜绿假单胞菌呈现中等活性(MIC = 250 μg/mL)。馏分 V 对枯草芽孢杆菌(MIC = 15.6 μg/mL)和金黄色葡萄球菌(MIC = 31.3 μg/mL)呈现高活性,对革兰氏阴性菌无活性(MIC > 1000 μg/mL)。然后馏分 III、IV 和 V 通过硅胶柱色谱进行生物测定导向的分级分离,得到单独纯化的多花萝芙木碱 A 和 B。两种多花萝芙木碱对金黄色葡萄球菌(MIC = 25 μg/mL)和粪肠球菌(MIC = 50 μg/mL)均显示出显著活性,多花萝芙木碱 A 和 B 对金黄色葡萄球菌的 EC₅₀分别为 8 和 2.5 μg/mL。这些结果很有前景,表明这些化合物对革兰氏阳性菌具有生物活性。

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