Gonçalves Jenifer Pendiuk, Dos Santos Maria Luiza Ferreira, Rossi Gustavo Rodrigues, Costa Gagosian Viviana Stephanie, de Oliveira Carolina Camargo
Laboratory of Inflammatory and Neoplasic Cells, Cell Biology Department of Biological Sciences, Universidade Federal do Paraná, Av Cel Francisco H dos Santos, s/n, CEP 81530-980, Curitiba, PR, Brazil.
Laboratory of Inflammatory and Neoplasic Cells, Cell Biology Department of Biological Sciences, Universidade Federal do Paraná, Av Cel Francisco H dos Santos, s/n, CEP 81530-980, Curitiba, PR, Brazil.
Homeopathy. 2017 Aug;106(3):171-180. doi: 10.1016/j.homp.2017.02.004. Epub 2017 Apr 8.
Zinc is an essential trace element necessary for life. Traditional and complementary medicines use zinc-based formulations to treat different classes of diseases. Basic research on homeopathic preparations of zinc are rare and there are a few published clinical cases describing its effects on patients. The use of cell-based models in drug screening is a reliable source of evidence.
We sought to investigate experimental end-points using cell-based models to determine the effects of dilutions of Zincum metallicum prepared according to the Brazilian Homeopathic Pharmacopoeia. Murine RAW 264.7 macrophages and melanoma B16-F10 cell lines were cultured according to standard procedures. Cells were treated with either 5c, 6c or 30c Zincum metallicum and control cells with its respective vehicle (5c, 6c, or 30c Lactose). Macrophage activation by CD54 immunolabeling and intracellular reactive oxygen species (ROS) using DCFH-DA (2,7-dichlorodihydrofluorescein diacetate) were detected by flow cytometry. Phagocytic capacity (endocytic index) was quantified by light microscopy. Features of melanoma cells were analyzed by colorimetric assays to determine melanin content and cell proliferation rate. All obtained data were submitted to normality test followed by statistical analysis.
Zincum metallicum 6c shifted high ROS-producing macrophages to a low ROS-producing phenotype. Macrophage CD54 expression was increased by Zincum metallicum 5c. No changes in endocytic index were observed. Melanoma cells were not affected by any treatment we tested.
Differing responses and non-linearity were found on macrophages challenged with Zincum metallicum at high dilutions. No changes in melanoma cells were observed. Customised assays using target cells can be useful to investigate high-dilution effects. Other cell types and conditions should be explored.
锌是生命必需的微量元素。传统医学和补充替代医学使用含锌制剂治疗各类疾病。关于锌顺势疗法制剂的基础研究很少，仅有少数已发表的临床病例描述了其对患者的影响。在药物筛选中使用基于细胞的模型是可靠的证据来源。
我们试图使用基于细胞的模型研究实验终点，以确定根据巴西顺势疗法药典制备的金属锌稀释液的效果。按照标准程序培养小鼠RAW 264.7巨噬细胞和黑色素瘤B16-F10细胞系。细胞分别用5c、6c或30c金属锌处理，对照细胞用各自的赋形剂（5c、6c或30c乳糖）处理。通过流式细胞术检测CD54免疫标记激活巨噬细胞以及使用DCFH-DA（2,7-二氯二氢荧光素二乙酸酯）检测细胞内活性氧（ROS）。通过光学显微镜定量吞噬能力（内吞指数）。通过比色法分析黑色素瘤细胞的特征，以确定黑色素含量和细胞增殖率。所有获得的数据均进行正态性检验，然后进行统计分析。
6c金属锌使高ROS产生的巨噬细胞转变为低ROS产生的表型。5c金属锌增加了巨噬细胞CD54的表达。未观察到内吞指数的变化。黑色素瘤细胞未受到我们测试的任何处理的影响。
在高稀释度下用金属锌攻击的巨噬细胞上发现了不同的反应和非线性。未观察到黑色素瘤细胞的变化。使用靶细胞的定制检测方法可能有助于研究高稀释度效应。应探索其他细胞类型和条件。
