Shirasuna Koumei, Matsumoto Haruka, Kobayashi Eiji, Nitta Akane, Haneda Shingo, Matsui Motozumi, Kawashima Chiho, Kida Katsuya, Shimizu Takashi, Miyamoto Akio
Graduate School of Animal and Food Hygiene, Obihiro University of Agriculture and Veterinary Medicine, Obihiro 080-8555, Japan.
J Reprod Dev. 2012;58(1):84-90. doi: 10.1262/jrd.11-094k. Epub 2011 Nov 4.
In cows, interferon-tau (IFNT) regulates maternal recognition around days 15-19 after artificial insemination (AI). The present study hypothesized that if key target genes of IFNT are clearly upregulated in earlier stages of pregnancy, these genes could be use as indices of future pregnancy in cows. Therefore, we determined the expression of these genes in peripheral blood mononuclear leukocytes (PBMCs) and polymorphonuclear granulocytes (PMNs) during the maternal recognition period (MRP). Twenty multiparous Holstein cows were subjected to AI on day 0 and categorized into the following groups: pregnancy (Preg, n = 9), embryonic death (ED, n = 5) and non-pregnancy (NP, n = 6). Progesterone levels in the Preg group were higher than those in the NP group on days 12-21. ISG15 and OAS-1 (IFN-stimulated genes: ISGs) mRNA in PBMCs on day 8 was higher in the Preg group than in the NP group, and these mRNAs in PMNs was higher in the Preg group on day 5 than in the NP and ED groups. Interleukin-10 (IL-10, Th2 cytokine) mRNA expression increased on day 8 in the PBMCs of pregnant cows. Tumor necrosis factor α (TNFα, Th1 cytokine) mRNA expression was stable in all groups. In an in vitro cell culture experiment, IFNT stimulated mRNA expression of ISGs in both PBMCs and PMNs. IFNT stimulated IL-10 mRNA expression in PBMCs, whereas IFNT increased TNFα mRNA levels in PBMCs in vitro. The results suggest that ISGs and IL-10 could be responsive to IFNT before the MRP in peripheral blood immune cells and may be useful target genes for reliable indices of pregnancy before the MRP.
在奶牛中,干扰素τ(IFNT)在人工授精(AI)后第15 - 19天左右调节母体识别。本研究假设,如果IFNT的关键靶基因在妊娠早期明显上调,这些基因可作为奶牛未来妊娠的指标。因此,我们测定了这些基因在母体识别期(MRP)外周血单个核白细胞(PBMC)和多形核粒细胞(PMN)中的表达。20头经产荷斯坦奶牛在第0天接受AI,并分为以下几组:妊娠组(Preg,n = 9)、胚胎死亡组(ED,n = 5)和非妊娠组(NP,n = 6)。妊娠组在第12 - 天的孕酮水平高于非妊娠组。妊娠组第8天PBMC中的ISG15和OAS - 1(干扰素刺激基因:ISGs)mRNA高于非妊娠组,妊娠组第5天PMN中的这些mRNA高于非妊娠组和胚胎死亡组。妊娠奶牛PBMC中白细胞介素10(IL - 10,Th2细胞因子)mRNA表达在第8天增加。肿瘤坏死因子α(TNFα,Th1细胞因子)mRNA表达在所有组中稳定。在体外细胞培养实验中,IFNT刺激PBMC和PMN中ISGs的mRNA表达。IFNT刺激PBMC中IL - 10 mRNA表达,而IFNT在体外增加PBMC中TNFα mRNA水平。结果表明,ISGs和IL - 10可能在MRP之前对外周血免疫细胞中的IFNT有反应,并且可能是MRP之前可靠妊娠指标的有用靶基因。