Dipartimento di Medicina Veterinaria, Università degli Studi di Milano, 20133, Milan, Italy.
Department of Biotechnology and Health Sciences, Molecular Biotechnology Center, Università di Torino, 10126, Turin, Italy.
Sci Rep. 2021 Jul 19;11(1):14712. doi: 10.1038/s41598-021-94113-8.
Post-surgical management is an important issue in veterinary medicine, requiring biomarkers with high sensitivity and specificity for timely and effective treatment. Emerging evidence suggests that miRNAs are promising stress- and pain-related markers. The aims were to profile the circulating miRNA signature in plasma of turtles (Trachemys scripta) and point out potential candidate biomarkers to assess the status of the animal. The plasma of female turtles underwent surgical gonadectomy were collected 24 h pre-surgery, and 2.5 h and 36 h post-surgery. The expression of miRNAs was profiled by Next Generation Sequencing and the dysregulated miRNAs were validated using RT-qPCR. The diagnostic value of miRNAs was calculated by ROC curves. The results showed that 14 miRNAs were differentially expressed over time. RT-qPCR validation highlighted that 2-miR-499-3p and miR-203-5p-out of 8 miRNAs tested were effectively modulated. The Area Under the Curve (AUC) of miR-203-5p was fair (AUC 0.7934) in discriminating pre- and 36 h post-surgery samples and poor for other time points; the AUC of miR-499-3p was excellent (AUC 0.944) in discriminating pre-surgery and 2.5 h post-surgery samples, and fair in discriminating pre-surgery and 36 h post-surgery (AUC 0.7292) and 2.5 h and 36 h post-surgery (AUC 0.7569) samples. In conclusion, we demonstrated for the first time that miRNAs profile changes in plasma of turtles underwent surgical oophorectomy and identified miR-203-5p and miR-499-3p as potential candidate biomarkers to assess animals' status. Further studies are necessary to confirm their diagnostic value and to investigate functional and mechanistic networks to improve our understanding of the biological processes.
手术后管理是兽医医学中的一个重要问题，需要具有高灵敏度和特异性的生物标志物，以便进行及时有效的治疗。新出现的证据表明，miRNA 是有前途的应激和疼痛相关标志物。本研究旨在分析龟（Trachemys scripta）血浆中循环 miRNA 特征，并指出潜在的候选生物标志物，以评估动物的状态。采集了接受手术去势的雌性龟的血浆，分别在术前 24 小时、术后 2.5 小时和 36 小时采集。通过下一代测序对 miRNA 的表达进行了分析，并使用 RT-qPCR 对失调的 miRNA 进行了验证。通过 ROC 曲线计算了 miRNA 的诊断价值。结果表明，有 14 个 miRNA 随时间表现出差异表达。在 8 个测试的 miRNA 中，RT-qPCR 验证显示 2-miR-499-3p 和 miR-203-5p 被有效调节。miR-203-5p 的曲线下面积（AUC）在区分术前和术后 36 小时样本时为中等（AUC 0.7934），在区分其他时间点时较差；miR-499-3p 的 AUC 在区分术前和术后 2.5 小时样本时极好（AUC 0.944），在区分术前和术后 36 小时（AUC 0.7292）和术前和术后 2.5 小时和 36 小时（AUC 0.7569）样本时为中等。总之，本研究首次证明了 miRNA 谱在接受手术去势的龟血浆中发生变化，并确定了 miR-203-5p 和 miR-499-3p 作为评估动物状态的潜在候选生物标志物。需要进一步的研究来确认它们的诊断价值，并研究功能和机制网络，以提高我们对生物学过程的理解。
