评估金枪鱼中异雌激素的生物测定试验和质谱法

Nader Akbari , Shahrbano Rostami , Mahmoud Ghazi-Khansari , Gholamreza Jahed-Khaniki , Nabi Shariatifar , Parisa Sadighara
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摘要

通过食物摄入异雌激素是一个重大问题。这些化合物具有类似雌激素的活性,会导致细胞增殖。本研究利用 MCF-7 细胞对罐头金枪鱼提取物进行生物测定和细胞增殖评估。首先,使用 MTT 试验对金枪鱼罐头的固体和液体部分进行了预筛选。结果发现,金枪鱼罐头固体部分的细胞增殖实际上发生在金枪鱼肉中。因此,我们选择了这部分金枪鱼罐头进行进一步研究。通过 RT-qPCR 确定了与 ERα 和 ERβ 受体的结合亲和力,并通过质谱分析确定了提取物中的雌激素成分。研究显示,使用金枪鱼罐头提取物处理后,细胞增殖率增加。17β-雌二醇被视为阳性对照。与 17β-estradiol 相比,细胞增殖率从 27% 到 36.6% 不等。在样品提取物组中,MCF-7 细胞的 ERα 和 ERβ 受体受到的刺激几乎相同。质谱分析确定了双酚 A(BPA)和其他雌激素化合物,如植物雌激素、邻苯二甲酸盐、多氯联苯、天然和合成雌激素。我们的研究结果表明,有必要结合使用生物测定试验和分析方法,以仔细管理和评估食品样本中各种成分对雌激素的综合影响。
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The bioassay tests and mass spectrometry for evaluation of xenoestrogens in tuna fish
Exposure to xenoestrogens through food is a significant concern. These compounds have estrogen-like activity and lead to cell proliferation. In this study, MCF-7 cells were utilized for bioassay tests and evaluation of cell proliferation of extracts prepared from canned tuna fish. First, a pre-screening using the MTT test was done with the solid and liquid parts of the canned tuna fish. It was observed that the proliferation of cells in the solid part of canned tuna cans actually occurs in tuna meat. Therefore, this portion of canned tuna fish was selected for further studies. The binding affinity with ERα and ERβ receptors was determined through RT-qPCR, and the extract was analyzed via mass spectrometry to identify estrogenic components. The study revealed an increased cell proliferation rate upon treatment with the canned tuna fish extract. 17β-estradiol was considered as positive control. The cell proliferation rate ranged from 27 % to 36.6 % compared to 17β-estradiol. Both ERα and ERβ receptors of MCF-7 cells were stimulated almost equally in the sample extract group. Mass spectrometry analysis identified bisphenol A(BPA) and other estrogenic compounds such as phytoestrogens, phthalates, polychlorinated biphenyls, natural and synthetic estrogen hormones. Our findings emphasize the necessity of using a combination of bioassay tests and analytical methods to carefully manage and evaluate the combined estrogenic effects of components in food samples.
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