这项研究的目的是确定妊娠早期水牛母体循环中妊娠相关糖蛋白-1(PAG-1)mRNA表达的存在。同时,干扰素-tau(IFNt)和一些干扰素刺激基因(ISGs)的mRNA表达水平(干扰素刺激基因15泛素样修饰干扰素,ISG15;混合病毒抗性1和2,MX1和MX2;2',对5'-寡腺苷酸合酶1,OAS1)进行了评估,以扩大我们对怀孕早期阶段涉及的分子过程的了解,并鉴定水牛中母胎细胞相互作用的潜在生物标志物。这项研究是在38头同步和人工授精的水牛(d0)上进行的,事后分为3组:怀孕(n=17),非妊娠(n=15)和胚胎死亡率(n=6)。在人工授精(AI)后的第14、19、28和40天收集血液样品以分离外周血单核细胞(PBMC)。PAG-1、IFNt、ISG15.使用RT-qPCR测量MX1、MX2和OAS1。各组间IFNt和PAG基因表达无明显变化,而ISG15、MX1、MX2和OAS1存在显著差异(p<0.001)。配对比较显示,两组之间的差异发生在AI后第19天和第28天。通过ROC分析,ISG15被证明具有最佳的诊断性能,可以区分怀孕动物和经历胚胎死亡的动物。根据单变量分析的结果,第19天被确定为区分组之间的最具指示性的基因,而这种分化的最可靠基因是ISG15,MX1和MX2。MX2被证明是使用判别分析区分怀孕水牛的最佳基因,而MX1是最好的预测胚胎死亡率的基因。我们的结果表明,在PAG-1中,IFNt和ISGs的表达可作为水牛母胎细胞相互作用的诊断和预后标志物,ISGs被证明是预测围植入期妊娠和胚胎死亡率的最佳外周生物标志物。这些对母胎相互作用背后的机制的见解以及早期发现胚胎窘迫的方法的发展可能使我们能够实施有效的策略来支持胚胎存活。
The aim of this study was to determine the presence of Pregnancy-associated glycoprotein -1 (PAG-1) mRNA expression in the maternal circulation of pregnant buffaloes during the early stage of pregnancy. Contemporaneously, the mRNA expression levels of Interferon-tau (
IFNt) and some Interferon stimulated genes (ISGs) (interferon stimulated gene 15 ubiquitin-like modifier interferon, ISG15; Mixoviruses resistance 1 and 2, MX1 and MX2; 2\',5\'-oligoadenylate synthase 1,OAS1) were evaluated in order to expand our knowledge of the molecular processes involved in the early stages of pregnancy and to identify potential biomarkers of maternal-fetal cellular interaction in buffalo. The study was conducted on 38 synchronized and artificially inseminated buffalo cows (d 0), divided ex post into 3 groups: Pregnant (n = 17), Non-pregnant (n = 15) and Embryo mortality (n = 6). Blood samples were collected on d 14, 19, 28 and 40 after artificial insemination (AI) for peripheral blood mononuclear cells (PBMCs) isolation. Expression levels of mRNA of PAG-1,
IFNt, ISG15. MX1, MX2 and OAS1 were measured using RT-qPCR. No significant changes were observed in
IFNt and PAG gene expressions between groups, while significant differences (p < 0.001) were found for ISG15, MX1, MX2, and OAS1. Pairwise comparisons revealed that the differences between groups occurred on days 19 and 28 post-AI. ISG15 proved to have the best diagnostic performance for distinguishing between pregnant animals and animals that experienced embryo mortality with the ROC analysis. According to the results of the univariate analyses, day 19 was identified as the most indicative to discriminate between groups while the most reliable genes for this differentiation were ISG15, MX1 and MX2. MX2 proved to be the best gene for discriminating pregnant buffaloes using the discriminant analysis, while MX1 was the gene that best predicted embryo mortality. Our results showed that among PAG-1,
IFNt and ISGs expression as diagnostic and prognostic markers of maternal-fetal cellular interaction in buffalo cows, ISGs proved to be the best peripheral biomarkers for predicting pregnancy and embryonic mortality during the peri-implantation period. These insights into the mechanisms behind maternal-fetal interaction and the development of a method for the early detection of embryo distress may enable us to implement effective strategies to support embryo survival.