Waller Stefanie B, Peter Cristina M, Hoffmann Jéssica F, Picoli Tony, Osório Luiza da G, Chaves Fábio, Zani João L, de Faria Renata O, de Mello João R B, Meireles Mário C A
Centro de Diagnóstico e Pesquisa em Micologia Veterinária, Universidade Federal de Pelotas, Pelotas, RS, Brazil.
Laboratório de Bacteriologia e Saúde Populacional, Universidade Federal de Pelotas, Pelotas, RS, Brazil.
Microb Pathog. 2017 Apr;105:117-121. doi: 10.1016/j.micpath.2017.02.022. Epub 2017 Feb 17.
This study aimed to evaluate the chemical composition and cytotoxic activity of brown Brazilian propolis and its in vitro activity against itraconazole-resistant Sporothrix brasiliensis from animal sporotrichosis. Propolis was acquired commercially and prepared as a hydroalcoholic extract. Chemical analysis was evaluated by liquid chromatography coupled to mass spectrometry of ultra-efficiency. The cell viability was evaluated by MTT test in MDBK cells of 50 to 0.09 μg/mL. For antifungal tests, twenty isolates of Sporothrix brasiliensis from dogs (n = 11) and cats (n = 9) with sporotrichosis were tested to itraconazole (16-0.0313 μg/mL) and to propolis (3.125-0.09 mg/mL) by broth microdilution technique (CLSI M38-A2), adapted to natural products. The results were expressed in minimal inhibitory concentration (MIC) and minimal fungicidal concentration (MFC). Itraconazole showed activity between MIC values of 0.25 to greater than 16 μg/mL, and 88.9% (08/09) and 72.7% (08/11) of S. brasiliensis from cats and dogs, respectively, were considered itraconazole-resistant. All Sporothrix brasiliensis were sensitive to brown propolis between MIC values of 0.19-1.56 mg/mL, including the itraconazole-resistant isolates, whereas the MFC values of propolis were from 0.78 to greater than 3.125 mg/mL. Propolis maintained a medium to high cell viability between concentration of 0.78 to 0.09 μg/mL, and p-coumaric acid was the major compound. Brown Brazilian propolis is a promising antifungal candidate against sporotrichosis and more studies need to be undertaken to evaluate its safe use to understand its efficacy.
本研究旨在评估巴西棕蜂胶的化学成分和细胞毒性活性，以及其对动物孢子丝菌病中耐伊曲康唑的巴西孢子丝菌的体外活性。蜂胶通过商业途径购得，并制备成水醇提取物。通过超高效液相色谱-质谱联用进行化学分析。采用MTT试验在浓度为50至0.09μg/mL的MDBK细胞中评估细胞活力。对于抗真菌试验，采用肉汤微量稀释技术（CLSI M38-A2，适用于天然产物），对从患有孢子丝菌病的犬（n = 11）和猫（n = 9）分离出的20株巴西孢子丝菌进行伊曲康唑（16 - 0.0313μg/mL）和蜂胶（3.125 - 0.09mg/mL）测试。结果以最低抑菌浓度（MIC）和最低杀菌浓度（MFC）表示。伊曲康唑的MIC值在0.25至大于16μg/mL之间，来自猫和犬的巴西孢子丝菌分别有88.9%（08/09）和72.7%（08/11）被认为对伊曲康唑耐药。所有巴西孢子丝菌对棕蜂胶敏感，MIC值在0.19 - 1.56mg/mL之间，包括耐伊曲康唑的分离株，而蜂胶的MFC值在0.78至大于3.125mg/mL之间。蜂胶在浓度为0.78至0.09μg/mL之间保持中等至高细胞活力，对香豆酸是主要成分。巴西棕蜂胶是一种有前景的抗孢子丝菌病的抗真菌候选物，需要进行更多研究以评估其安全使用情况并了解其疗效。
