Guzeloglu A, Michel F, Thatcher W W
Department of Animal Sciences, University of Florida, Gainesville 32611, USA.
J Dairy Sci. 2004 Jul;87(7):2032-41. doi: 10.3168/jds.S0022-0302(04)70021-1.
Rationale for these experiments was to evaluate the dose effects of bovine interferon-tau (IFN-tau) on the prostaglandin secretory pathway of immortalized bovine endometrial (BEND) cells in response to phorbol 12, 13-dibutyrate (PdBu) and to characterize similar responses in primary bovine uterine epithelial cells as a biomonitor of embryo-induced antiluteolytic effects on the endometrium. The BEND cells were treated with PdBu (0 or 100 ng/mL) and IFN-tau (0 or 50 ng/mL) for 6 h. The PdBu stimulated secretions of PGF2alpha and prostaglandin E2 (PGE2). Co-treatment of cells with IFN-tau blocked PdBu-induced secretion of both PGF2alpha and PGE2. Treatment with PdBu for 6 h induced expression of prostaglandin H synthase-2 mRNA, prostaglandin H synthase-2 protein, and prostaglandin E synthase mRNA, which were blocked with concurrent addition of IFN-tau. Doses of IFN-tau (0.05, 0.5, 1, 5, 10, and 20 microg/mL) were used with PdBu (0 and 100 ng/mL). The IFN-tau alone failed to stimulate secretion of PGF2alpha and PGE2, whereas IFN-tau doses <5 microg/mL suppressed PdBu-stimulated secretions of PGF2alpha and PGE2. Uterine epithelial cells were isolated from cows at d 17 after estrus and were cultured to confluence in serum-free medium. Cells were treated with IFN-tau (0, 50, or 500 ng/mL) and PdBu (0 or 100 ng/mL) before media were collected after 24 h for PGF2alpha and PGE2 analyses. Treatment of primary uterine epithelial cells with PdBu induced PGF2alpha secretion, and IFN-tau (50 and 500 ng/mL) caused a reduction in PGF2alpha secretion induced by PdBu. In the absence of PdBu, IFN-tau increased basal secretion of PGF2alpha. Concentrations of PGE2 increased in response to PdBu, and the 50-ng/mL dose of IFN-tau had a stimulatory effect on PGE2 concentrations compared with the 500-ng/mL dose in the absence of PdBu. Phorbol ester-induced gene transcription as related to prostaglandin synthesis is regulated by IFN-tau in vitro.
这些实验的目的是评估牛干扰素 - τ（IFN - τ）对永生化牛子宫内膜（BEND）细胞前列腺素分泌途径的剂量效应，该效应是对佛波酯12,13 - 二丁酸酯（PdBu）的反应，并将原代牛子宫上皮细胞中的类似反应作为胚胎诱导的子宫内膜抗溶解作用的生物监测指标进行表征。将BEND细胞用PdBu（0或100 ng/mL）和IFN - τ（0或50 ng/mL）处理6小时。PdBu刺激了前列腺素F2α（PGF2α）和前列腺素E2（PGE2）的分泌。IFN - τ与细胞共同处理可阻断PdBu诱导的PGF2α和PGE2的分泌。用PdBu处理6小时可诱导前列腺素H合酶 - 2 mRNA、前列腺素H合酶 - 2蛋白和前列腺素E合酶mRNA的表达，同时添加IFN - τ可阻断这些表达。使用IFN - τ剂量（0.05、0.5、1、5、10和20 μg/mL）与PdBu（0和100 ng/mL）一起使用。单独的IFN - τ未能刺激PGF2α和PGE2的分泌，而IFN - τ剂量<5 μg/mL可抑制PdBu刺激的PGF2α和PGE2的分泌。在发情后第17天从奶牛分离子宫上皮细胞，并在无血清培养基中培养至汇合。在收集培养基24小时后进行PGF2α和PGE2分析之前，将细胞用IFN - τ（0、50或500 ng/mL）和PdBu（0或100 ng/mL）处理。用PdBu处理原代子宫上皮细胞可诱导PGF2α分泌，而IFN - τ（50和500 ng/mL）可导致PdBu诱导的PGF2α分泌减少。在没有PdBu的情况下，IFN - τ增加了PGF2α的基础分泌。PGE2的浓度因PdBu而增加，与在没有PdBu的情况下500 ng/mL剂量相比，50 ng/mL剂量的IFN - τ对PGE2浓度有刺激作用。在体外，佛波酯诱导的与前列腺素合成相关的基因转录受IFN - τ调节。
