de Mendonça Izabel Cristina Gomes, Porto Isabel Cristina Celerino de Moraes, do Nascimento Ticiano Gomes, de Souza Naiana Soares, Oliveira José Marcos dos Santos, Arruda Rodolfo Elleson dos Santos, Mousinho Kristiana Cerqueira, dos Santos Aldenir Feitosa, Basílio-Júnior Irinaldo Diniz, Parolia Abhishek, Barreto Francisco Stefânio
School of Dentistry, Cesmac University Center, Rua Cônego Machado, 918, Farol, CEP: 57051-160, Maceió, Alagoas, Brazil.
School of Nursing and Pharmacy, Federal University of Alagoas, Campus A. C. Simões, University City, Maceió, 57072-970 , AL, Brazil.
BMC Complement Altern Med. 2015 Oct 14;15:357. doi: 10.1186/s12906-015-0888-9.
The implementation of new public healthcare models that stimulate the use of natural products from traditional medicine, as a so-called integrated medicine, refers to an approach that use best of both conventional medicine and traditional medicine. Propolis is a widely used natural product by different ancient cultures and known to exhibit biological activities beneficial for health. The large number of studies conducted with propolis had shown that its chemical composition differs as a function of the climate, plant diversity and bee species and plays an important role on its therapeutic properties. The aim of this study was to analyse the phytochemical profile of the ethanolic extract of red propolis (EEP) and its fractionation, antioxidant action of EEP and its fractions hexane, cloroform and ethyl acetate and cytotoxic activity of EEP on human tumour cell lines SF-295 (glioblastoma), OVCAR-8 (ovary) and HCT-116 (colon).
EEP was obtained by maceration with absolute ethanol, then it was concentrated in rotaevaporator up to complete evaporation of the solvent. The crude extract was fractionated with hexane, ethyl acetate, chloroform and methanol and they were subjected to phytochemical screening and total phenolic compounds. Antioxidant activity of EEP and fractions was done by means of the 2,2-diphenyl-1-picryhydrazyl (DPPH) method. Biomarkers of red propolis were identified by LC-Orbitrap-FTMS. To assess cytotoxic activity of the extract, cells were exposed to EEP over 72 h. Cell viability was assessed by means of MTT assay. The percentage of cell growth inhibition (IC50) was analysed by means of non-linear regression, and the absorbance values of the various investigated concentrations were subjected to one-factor analysis of variance (ANOVA) followed by Tukey's or Tamhane's tests (α = 0.05).
The results obtained using phytochemical screening and LC-Orbitrap-FTMS indicated the presence of phlobaphene tannins, catechins, chalcones, aurones, flavonones, flavonols, xanthones, pentacyclic triterpenoids and guttiferones in Brazilian red propolis. EEP and its hexane, chloroform and ethyl acetate fractions obtained by liquid-liquid partitioning exhibited satisfactory antioxidant percentages. EEP (IC50 < 34.27 μg/mL) exhibited high levels of cytotoxicity on all human tumour cell lines tested when compared to negative control.
C-Orbitrap-FTMS was useful to establish the chemical profile of the red propolis. Brazilian red propolis has antioxidant properties and decreases substantially the percentage of cell survival of human tumour cells; thus, it has potential to serve as an anticancer drug.
新的公共医疗模式的实施鼓励使用传统医学中的天然产品，即所谓的整合医学，它融合了传统医学和现代医学的优势。蜂胶是一种在不同古代文化中广泛使用的天然产品，具有多种对健康有益的生物活性。大量关于蜂胶的研究表明，其化学成分因气候、植物多样性和蜜蜂种类的不同而有所差异，这对其治疗特性起着重要作用。本研究旨在分析红蜂胶乙醇提取物（EEP）的植物化学特征及其分离情况，研究EEP及其己烷、氯仿和乙酸乙酯馏分的抗氧化作用，以及EEP对人肿瘤细胞系SF - 295（胶质母细胞瘤）、OVCAR - 8（卵巢癌）和HCT - 116（结肠癌）的细胞毒性活性。
通过用无水乙醇浸渍获得EEP，然后在旋转蒸发仪中浓缩直至溶剂完全蒸发。粗提取物用己烷、乙酸乙酯、氯仿和甲醇进行分离，并对它们进行植物化学筛选和总酚类化合物分析。EEP及其馏分的抗氧化活性通过2,2 - 二苯基 - 1 - 苦基肼（DPPH）法测定。通过液相色谱 - 轨道阱 - 傅里叶变换质谱仪（LC - Orbitrap - FTMS）鉴定红蜂胶的生物标志物。为了评估提取物的细胞毒性活性，将细胞暴露于EEP中72小时。通过MTT法评估细胞活力。通过非线性回归分析细胞生长抑制百分比（IC50），对各种研究浓度的吸光度值进行单因素方差分析（ANOVA），然后进行Tukey检验或Tamhane检验（α = 0.05）。
植物化学筛选和LC - Orbitrap - FTMS的结果表明，巴西红蜂胶中存在鞣红单宁、儿茶素、查耳酮、噢哢、黄酮酮、黄酮醇、呫吨酮、五环三萜和藤黄萘醌。通过液 - 液分配获得的EEP及其己烷、氯仿和乙酸乙酯馏分表现出令人满意的抗氧化百分比。与阴性对照相比，EEP（IC50 < 34.27 μg/mL）对所有测试的人肿瘤细胞系均表现出高水平的细胞毒性。
LC - Orbitrap - FTMS有助于确定红蜂胶的化学特征。巴西红蜂胶具有抗氧化特性，能显著降低人肿瘤细胞的细胞存活百分比；因此，它有潜力作为一种抗癌药物。
