Solomando Juan Carlos, Antequera Teresa, Estaras Matías, González Antonio, Perez-Palacios Trinidad
Research Institute of Meat and Meat Products (IProCar) University of Extremadura Cáceres Spain.
Department of Physiology, Institute of Biomarkers and Molecular Pathologies University of Extremadura Cáceres Spain.
Food Sci Nutr. 2024 Jun 17;12(9):6338-6352. doi: 10.1002/fsn3.4241. eCollection 2024 Sep.
This work aimed to evaluate the Caco-2 cells as a model to study the epithelial transport of intestinal lipid extracts subjected to in vitro digestion, to establish a standard protocol for the determination of bioaccessibility and bioavailability of fatty acids in meat matrix, especially in those enriched with ω-3 (eicosapentaenoic (EPA) and docosahexaenoic acids (DHA)). Samples were first subjected to in vitro digestion, and then, the intestinal extract was incubated with Caco-2 cells. A first trial was conducted to select the most influencing variables on the fatty acid transport during Caco-2 cell incubation: fat quantity on the intestinal extract, incubation time, and shaking. Then, a second experiment was carried out to determine the influence of these variables, being the fat quantity and the incubation time the most influencing factors on the transport and bioavailability of fatty acids. The effect of shaking was not so marked but seemed to improve the bioavailability of saturated fatty acids. This study also allows establishing the most suitable conditions: intestinal extracts with 30 mg of fat, longer incubation times (8 h), and shaking, achieving active and passive fatty acid transport without compromising the integrity of the Caco-2 cell monolayer. The accurate results obtained for major and minor fatty acids, especially EPA and DHA are remarkable, due to the interest in these bioactive compounds. Thus, this study provides a combined protocol based on static in vitro digestion followed by Caco-2 cell incubation to assess the bioaccessibility and bioavailability of fatty acids in meat samples.
本研究旨在评估将Caco-2细胞作为模型，用于研究体外消化后的肠道脂质提取物的上皮转运，建立一种用于测定肉类基质中脂肪酸生物可及性和生物利用度的标准方案，特别是在富含ω-3（二十碳五烯酸（EPA）和二十二碳六烯酸（DHA））的肉类基质中。样品首先进行体外消化，然后将肠道提取物与Caco-2细胞孵育。进行了第一项试验，以选择在Caco-2细胞孵育过程中对脂肪酸转运影响最大的变量：肠道提取物中的脂肪量、孵育时间和振荡。然后，进行了第二项实验，以确定这些变量的影响，其中脂肪量和孵育时间是对脂肪酸转运和生物利用度影响最大的因素。振荡的影响不太明显，但似乎提高了饱和脂肪酸的生物利用度。本研究还确定了最合适的条件：含有30毫克脂肪的肠道提取物、更长的孵育时间（8小时）和振荡，在不损害Caco-2细胞单层完整性的情况下实现脂肪酸的主动和被动转运。由于对这些生物活性化合物的关注，对于主要和次要脂肪酸，特别是EPA和DHA所获得的准确结果非常显著。因此，本研究提供了一种基于静态体外消化然后进行Caco-2细胞孵育的联合方案，以评估肉类样品中脂肪酸的生物可及性和生物利用度。
