Poit Diego Angelo Schmidt, Ferraz Priscila Assis, Bruni Gabriela de Andrade, Bruni Giulia de Andrade, Nishimura Thiago Kan, Motta Igor Garcia, Feltrin Isabella Rio, Pugliesi Guilherme
Departamento de Reprodução Animal, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Pirassununga, SP, Brasil.
Departamento de Produção Animal, Faculdade de Medicina Veterinária e Zootecnia, Universidade Estadual Paulista, Botucatu, SP, Brasil.
Anim Reprod. 2024 Jul 15;21(2):e20230144. doi: 10.1590/1984-3143-AR2023-0144. eCollection 2024.
In Experiment 1, PBMC were isolated from cows considered healthy or with SCE (n=6/group) on Days 0 (estrus) and 7 (diestrus) of a synchronized estrous cycle. In Experiment 2, on D21 (D0 was defined as the day of Fixed Timed Artificial Insemination (FTAI), cows were evaluated by ultrasonography to assess luteal blood perfusion and PBMC were isolated. On D32, cows were classified into: healthy pregnant (n=7), pregnant with SCE (n=4), healthy non-pregnant (n=8), and non-pregnant with SCE (n=10). Gene expression of ISGs (ISG15, OAS1, MX1 and IFI6) and proinflammatory cytokines (IL1-β, TNF-α and IFN-γ) were determined. Expression of ISG15, MX1, IFI6, TNF-α and IFN-γ did not differ between SCE and healthy cows and between Days 0 and 7. Expression of OAS1 and IL1-β were higher (P=0.02) on Day 7 than Day 0, regardlees of the SCE presence. In Exp.2, ISG15 abundance was 2.5-fold greater (P=0.0008), TNF-α was 2.2-fold greater (P=0.05), and IL1-β tended (P=0.06) to be 2.4-fold higher in pregnant than non-pregnant cows. Luteal blood perfusion was greater (P=0.01) in pregnant animals. In conclusion, OAS1 and IL1-β are transcripts upregulated in PBMC at diestrus, regardless of SCE occurrence. Proinflammatory cytokines are not affected by SCE occurrence, but IL1-β and TNF-α are upregulated in pregnant animals on D21 of pregnancy. ISG15 abundance is a good pregnancy predictor, regardless SCE presence.
在实验1中，于同步发情周期的第0天（发情期）和第7天（间情期）从被认为健康或患有亚临床子宫内膜炎（SCE）的奶牛（每组n = 6）中分离外周血单核细胞（PBMC）。在实验2中，于第21天（将第0天定义为定时人工授精（FTAI）日），通过超声检查评估奶牛的黄体血流灌注并分离PBMC。在第32天，奶牛被分为：健康怀孕（n = 7）、患有SCE的怀孕（n = 4）、健康未怀孕（n = 8）和患有SCE的未怀孕（n = 10）。测定了干扰素刺激基因（ISG，即ISG15、OAS1、MX1和IFI6）和促炎细胞因子（IL1-β、TNF-α和IFN-γ）的基因表达。SCE奶牛与健康奶牛之间以及第0天和第7天之间，ISG15、MX1、IFI6、TNF-α和IFN-γ的表达没有差异。无论是否存在SCE，OAS1和IL1-β在第7天的表达均高于第0天（P = 0.02）。在实验2中，怀孕奶牛的ISG15丰度高2.5倍（P = 0.0008），TNF-α高2.2倍（P = 0.05），IL1-β有升高趋势（P = 0.06），高2.4倍。怀孕动物的黄体血流灌注更大（P = 0.01）。总之，无论是否发生SCE，OAS1和IL1-β都是间情期PBMC中上调的转录本。促炎细胞因子不受SCE发生的影响，但在怀孕第21天，怀孕动物的IL1-β和TNF-α上调。无论是否存在SCE，ISG15丰度都是良好的怀孕预测指标。
