Freires I A, Queiroz V C P P, Furletti V F, Ikegaki M, de Alencar S M, Duarte M C T, Rosalen P L
Department of Physiological Sciences, Piracicaba Dental School, University of Campinas (UNICAMP), 13414-018 Piracicaba, SP, Brazil.
Research Center for Chemistry, Biology and Agriculture, University of Campinas, Campinas (UNICAMP), Box 6171, 13081-970 Campinas, SP, Brazil.
J Mycol Med. 2016 Jun;26(2):122-132. doi: 10.1016/j.mycmed.2016.01.003. Epub 2016 Feb 23.
Propolis is known to have biological properties against numerous microorganisms of clinical interest. This study aimed to determine the chemical composition and antifungal activity of Brazilian propolis (types 3 and 13) against Candida spp. and their effects on the morphology of preformed and mature Candida biofilms. Samples of propolis (3 and 13) collected by Apis mellifera honeybees were obtained from different regions in Brazil. Ethanolic extracts of propolis (EEP) were prepared, fractionated and submitted to chemical analysis by GC/MS. The extracts and their hexane, dichloromethane and ethyl acetate fractions were tested for their ability to inhibit Candida spp. (C. albicans, C. dubliniensis, C. glabrata, C. kruzei, C. tropicalis and C. parapsilosis) by determination of the minimum inhibitory and fungicidal concentrations (MIC/MFC). Additionally, their effects on morphology of preformed and mature biofilms were observed by scanning electron microscopy. The phenolic compounds p-coumaric acid, caffeic acid phenethyl ester (CAPE), kaempferol and quercetin were identified in the EEP-3 and its bioactive dichloromethane fraction; and isoflavonoids such as medicarpin, vestitol and formononetin were found in the EEP-13, and triterpenes in its bioactive hexane fraction. The EEP-3 and EEP-13 and their bioactive fractions showed MIC values ranging from 0.2 to 125μg/mL and MFC values between 125 and 500μg/mL. The EEP and fractions were predominantly fungistatic agents. All extracts and fractions disrupted biofilm structures at 500μg/mL and amorphous areas with cell damage were clearly observed in preformed and mature biofilms. Propolis types 3 and 13 have strong anti-Candida activity and should be considered as promising candidates to treat oral and systemic candidiasis.
已知蜂胶具有针对众多具有临床意义的微生物的生物学特性。本研究旨在确定巴西蜂胶（3型和13型）对念珠菌属的化学成分和抗真菌活性，以及它们对预先形成的和成熟的念珠菌生物膜形态的影响。由意大利蜜蜂采集的蜂胶（3型和13型）样本取自巴西不同地区。制备蜂胶乙醇提取物（EEP），进行分馏并通过气相色谱/质谱联用仪进行化学分析。通过测定最低抑菌浓度和杀菌浓度（MIC/MFC），测试提取物及其己烷、二氯甲烷和乙酸乙酯馏分抑制念珠菌属（白色念珠菌、都柏林念珠菌、光滑念珠菌、克鲁斯念珠菌、热带念珠菌和近平滑念珠菌）的能力。此外，通过扫描电子显微镜观察它们对预先形成的和成熟生物膜形态的影响。在EEP - 3及其生物活性二氯甲烷馏分中鉴定出酚类化合物对香豆酸、咖啡酸苯乙酯（CAPE）、山柰酚和槲皮素；在EEP - 13中发现了异黄酮类化合物如紫穗槐二氢黄酮、黄檀醇和芒柄花素，在其生物活性己烷馏分中发现了三萜类化合物。EEP - 3和EEP - 13及其生物活性馏分的MIC值范围为0.2至125μg/mL，MFC值在125至500μg/mL之间。EEP及其馏分主要是抑菌剂。所有提取物和馏分在500μg/mL时破坏生物膜结构，并且在预先形成的和成熟的生物膜中均清晰观察到有细胞损伤的无定形区域。3型和13型蜂胶具有很强的抗念珠菌活性，应被视为治疗口腔和全身性念珠菌病的有前景的候选药物。
