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从田野勿忘草地上部分提取的提取物和三萜皂苷的抗真菌活性及细胞毒性

Antifungal activity and cytotoxicity of extracts and triterpenoid saponins obtained from the aerial parts of Anagallis arvensis L.

作者信息

Soberón José R, Sgariglia Melina A, Pastoriza Ana C, Soruco Estela M, Jäger Sebastián N, Labadie Guillermo R, Sampietro Diego A, Vattuone Marta A

机构信息

Cátedra de Fitoquímica, Instituto de Estudios Farmacológicos "Dr. A.R. Sampietro", Facultad de Bioquímica, Química y Farmacia, Universidad Nacional de Tucumán, Ayacucho 471, T4000INI San Miguel de Tucumán, Tucumán, Argentina; Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Godoy Cruz 2290, C1425FQB Ciudad Autónoma de Buenos Aires, Argentina.

Cátedra de Fitoquímica, Instituto de Estudios Farmacológicos "Dr. A.R. Sampietro", Facultad de Bioquímica, Química y Farmacia, Universidad Nacional de Tucumán, Ayacucho 471, T4000INI San Miguel de Tucumán, Tucumán, Argentina; Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Godoy Cruz 2290, C1425FQB Ciudad Autónoma de Buenos Aires, Argentina.

出版信息

J Ethnopharmacol. 2017 May 5;203:233-240. doi: 10.1016/j.jep.2017.03.056. Epub 2017 Apr 4.

Abstract

ETHNOPHARMACOLOGICAL RELEVANCE

Anagallis arvensis L. (Primulaceae) is used in argentinean northwestern traditional medicine to treat fungal infections. We are reporting the isolation and identification of compounds with antifungal activity against human pathogenic yeast Candida albicans, and toxicity evaluation.

AIM OF THE STUDY

to study the antifungal activity of extracts and purified compounds obtained form A. arvensis aerial parts, alone and in combinations with fluconazole (FLU), and to study the toxicity of the active compounds.

MATERIALS AND METHODS

Disk diffusion assays were used to perform an activity-guided isolation of antifungal compounds from the aerial parts of A. arvensis. Broth dilution checkerboard and viable cell count assays were employed to determine the effects of samples and combinations of FLU + samples against Candida albicans. The chemical structures of active compounds were elucidated by spectroscopic analysis. Genotoxic and haemolytic effects of the isolated compounds were determined.

RESULTS

Four triterpenoid saponins (1-4) were identified. Anagallisin C (AnC), exerted the highest inhibitory activity among the assayed compounds against C. albicans reference strain (ATCC 10231), with MIC-0 =1µg/mL. The Fractional Inhibitory Concentration Index (FICI=0.129) indicated a synergistic effect between AnC (0.125µg/mL) and FLU (0.031µg/mL) against C. albicans ATCC 10231. AnC inhibited C. albicans 12-99 FLU resistant strain (MIC-0 =1µg/mL), and the FICI=0.188 indicated a synergistic effect between AnC (0.125µg/mL) and fluconazole (16µg/mL). The combination AnC+ FLU exerted fungicidal activity against both C. albicans strains. AnC exerted inhibitory activity against C. albicans ATCC 10231 sessile cells (MIC0=0.5µg/mL and MIC=1µg/mL) and against C. albicans 12-99 sessile cells (MIC0=0.75µg/mL and MIC=1.25µg/mL). AnC exerted haemolytic effect against human red blood cells at 15µg/mL and did not exerted genotoxic effect on Bacillus subtilis rec strains.

CONCLUSIONS

The antifungal activity and lack of genotoxic effects of AnC give support to the traditional use of A. arvensis as antifungal and makes AnC a compound of interest to expand the available antifungal drugs.

摘要

民族药理学相关性

田野水杨梅(报春花科)在阿根廷西北部传统医学中用于治疗真菌感染。我们报告了对具有抗人类致病酵母白色念珠菌活性的化合物的分离与鉴定以及毒性评估。

研究目的

研究从田野水杨梅地上部分获得的提取物和纯化化合物单独以及与氟康唑(FLU)联合使用时的抗真菌活性,并研究活性化合物的毒性。

材料与方法

采用纸片扩散法对田野水杨梅地上部分的抗真菌化合物进行活性导向分离。采用肉汤稀释棋盘法和活菌计数法测定样品以及氟康唑与样品组合对白色念珠菌的作用。通过光谱分析阐明活性化合物的化学结构。测定分离化合物的遗传毒性和溶血作用。

结果

鉴定出四种三萜皂苷(1 - 4)。在测定的化合物中,田野水杨梅苷C(AnC)对白色念珠菌参考菌株(ATCC 10231)的抑制活性最高,MIC - 0 = 1μg/mL。分数抑制浓度指数(FICI = 0.129)表明AnC(0.125μg/mL)与氟康唑(0.031μg/mL)联合对白色念珠菌ATCC 10231有协同作用。AnC对白色念珠菌12 - 99氟康唑耐药菌株有抑制作用(MIC - 0 = 1μg/mL),FICI = 0.188表明AnC(0.125μg/mL)与氟康唑(16μg/mL)联合有协同作用。AnC与氟康唑联合对两种白色念珠菌菌株均有杀菌活性。AnC对白色念珠菌ATCC 10231固着细胞有抑制活性(MIC0 = 0.5μg/mL,MIC = 1μg/mL),对白色念珠菌12 - 99固着细胞也有抑制活性(MIC0 = 0.75μg/mL,MIC = 1.25μg/mL)。AnC在15μg/mL时对人红细胞有溶血作用,对枯草芽孢杆菌rec菌株无遗传毒性作用。

结论

田野水杨梅苷C的抗真菌活性及无遗传毒性作用支持了田野水杨梅作为抗真菌药物的传统用途,并使田野水杨梅苷C成为一种有潜力扩展现有抗真菌药物的化合物。

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