Optimizing Fluorescein Isothiocyanate Dextran Measurement As a Biomarker in a 24-h Feed Restriction Model to Induce Gut Permeability in Broiler Chickens.
Optimizing Fluorescein Isothiocyanate Dextran Measurement As a Biomarker in a 24-h Feed Restriction Model to Induce Gut Permeability in Broiler Chickens.
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DOI：
10.3389/fvets.2017.00056
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发表时间：
2017
影响因子：
3.2
通讯作者：
Tellez G
中科院分区：
文献类型：
--
作者：
Baxter MFA;Merino-Guzman R;Latorre JD;Mahaffey BD;Yang Y;Teague KD;Graham LE;Wolfenden AD;Hernandez-Velasco X;Bielke LR;Hargis BM;Tellez G
异硫氰酸黄绿素葡聚糖（FITC-d）是用于测量紧密连接渗透性的3-5 kDa标记物。我们以前已经表明，肠道屏障功能可以受到不利影响的压力，消化不良的饮食，或饲料限制（FR），导致增加肠道炎症相关的渗透性。然而，目前的FITC-d方法的进一步优化调整是可能的，以提高未来结果的精度和有效性。本研究的目的是优化我们目前的模型，以获得对照组和处理组之间的更大差异，通过优化FITC-d测量作为24小时FR模型中的生物标志物，以诱导肉鸡的肠道通透性。进行了一项体外和四项体内独立实验。本研究的结果表明，通过增加FITC-d的剂量，（8.32 vs 4.16 mg/kg）;缩短血样采集时间（1 vs 2.5 h）;使用非FITC-d血清池作为空白，与先前使用的PBS进行比较;通过添加标准曲线设置检测限和修改软件的最佳灵敏度值，可以获得更加一致和可靠的结果。
Fluorescein isothiocyanate dextran (FITC-d) is a 3–5 kDa marker used to measure tight junction permeability. We have previously shown that intestinal barrier function can be adversely affected by stress, poorly digested diets, or feed restriction (FR), resulting in increased intestinal inflammation-associated permeability. However, further optimization adjustments of the current FITC-d methodology are possible to enhance precision and efficacy of results in future. The objective of the present study was to optimize our current model to obtain a larger difference between control and treated groups, by optimizing the FITC-d measurement as a biomarker in a 24-h FR model to induce gut permeability in broiler chickens. One in vitro and four in vivo independent experiments were conducted. The results of the present study suggest that by increasing the dose of FITC-d (8.32 versus 4.16 mg/kg); shortening the collection time of blood samples (1 versus 2.5 h); using a pool of non-FITC-d serum as a blank, compared to previously used PBS; adding a standard curve to set a limit of detection and modifying the software’s optimal sensitivity value, it was possible to obtain more consistent and reliable results.