U.S. Food and Drug Administration Center for Veterinary Medicine, Laurel, MD, 20708, United States.
U.S. Food and Drug Administration Center for Drug Evaluation and Research, Silver Spring, MD, 20903, United States.
Vet Immunol Immunopathol. 2021 Jul;237:110267. doi: 10.1016/j.vetimm.2021.110267. Epub 2021 May 10.
In veterinary medicine, inflammation in swine is evaluated principally by clinical signs. This method is often unreliable when assessing large animal populations because of inconsistent interpretations of clinical observations. This study examined whether changes in miRNA expression can predict the severity of the inflammatory response in swine after administration of Escherichia coli lipopolysaccharide (LPS). Whole blood from swine challenged with LPS at 0.125 μg/kg to 2.0 μg/kg body weight was collected at 0, 1, 3, and 8 h post LPS-challenge. Mature miRNAs were extracted from plasma and quantitative real-time-PCR (qRT-PCR) was used to evaluate the 84 most abundant swine miRNAs found in plasma. The miRNA changes in expression were assessed using the comparative CT Method (ΔΔCT method) for normalization with an exogenous control. The results revealed that expression of ssc-let-7e-5p, ssc-mir-22-3p, and ssc-miR-146a-5p were the most significantly changed miRNA over the time course. At 1 h post-LPS, ssc-let-7e-5p decreased as the LPS dosage levels increased from 0.125 to 1.0 μg/kg. Similarly, as the LPS doses increased from 0.125 to 0.5 μg/kg, ssc-miR-22-3p levels significantly decreased at 1 h post-LPS. In the 2.0 μg/kg LPS, ssc-miR-146a-5p levels increased between 0 and 3 h post-LPS; however, expression was downregulated with a 145 % decrease from 3 to 8 h. The three miRNA biomarkers suggest potentially useful surrogate endpoints for the evaluation of inflammatory and endotoxemia responses in swine.
在兽医领域，猪只的炎症主要通过临床症状进行评估。然而，当评估大型动物群体时，这种方法往往不可靠，因为临床观察结果的解释不一致。本研究探讨了在给予大肠杆菌脂多糖（LPS）后，miRNA 表达的变化是否可以预测猪只炎症反应的严重程度。用 0.125μg/kg 至 2.0μg/kg 体重 LPS 对猪进行攻毒，在 LPS 攻毒后 0、1、3 和 8 小时采集全血。从血浆中提取成熟 miRNA，并用定量实时 PCR（qRT-PCR）评估血浆中发现的 84 种最丰富的猪 miRNA。使用比较 CT 法（ΔΔCT 法）对 miRNA 表达的变化进行评估，并用外源性对照进行归一化。结果表明，在整个时间过程中，ssc-let-7e-5p、ssc-mir-22-3p 和 ssc-miR-146a-5p 的表达变化最为显著。在 LPS 攻毒后 1 小时，随着 LPS 剂量从 0.125μg/kg 增加到 1.0μg/kg，ssc-let-7e-5p 的表达减少。同样，随着 LPS 剂量从 0.125μg/kg 增加到 0.5μg/kg，ssc-miR-22-3p 的水平在 LPS 攻毒后 1 小时显著下降。在 2.0μg/kg LPS 中，ssc-miR-146a-5p 的水平在 LPS 攻毒后 0 至 3 小时之间增加；然而，在 3 至 8 小时之间，其表达被下调，下降了 145%。这三个 miRNA 生物标志物提示可能是评估猪炎症和内毒素血症反应的有用替代终点。
