12-氧代二十碳四烯酸是胎盘分离的候选信号，可激活基质金属蛋白酶并诱导牛滋养层细胞凋亡。,Animal Bioscience

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12-氧代二十碳四烯酸是胎盘分离的候选信号，可激活基质金属蛋白酶并诱导牛滋养层细胞凋亡。
Animal Bioscience ( IF 2.4 ) Pub Date : 2022-06-30 , DOI: 10.5713/ab.22.0097
Hachiro Kamada _{1,

2}

Affiliation

目的 12-oxo-5Z,8Z,10E,14Z-二十碳四烯酸 (12-KETE) 是花生四烯酸的一种代谢产物，是小牛分娩后胎盘分离的强烈候选信号。在本研究中，研究了 12-KETE 对牛滋养层细胞的影响，以确定其在分娩时在胎盘组中的功能。方法使用源自胚泡的牛滋养层细胞。它们与或不与来自牛胎盘和/或牛子宫上皮细胞的成纤维细胞共培养。将 12-KETE 添加到培养基中。结果牛滋养层细胞含有双核细胞和强烈表达的尾型同源框 2 (CDX-2) 基因。将 12-KETE 添加到没有饲养细胞的滋养层细胞集落或在成纤维细胞单层上添加 12-KETE 可诱导集落的快速脱落。加入 12-KETE 后，滋养层细胞发出由染料淬灭的胶原蛋白降解引起的强烈荧光，表明 12-KETE 激活了滋养层细胞的基质金属蛋白酶。脱落的细胞集落用 YOPRO-1 染色，但未用碘化丙锭 (PI) 染色。此外，在脱落细胞中观察到 DNA 片段化和 Bcl-2 相关 X 蛋白 (Bax) 基因（凋亡刺激因子）上调，表明 12-KETE 诱导滋养层细胞凋亡。这些结果与之前的体内观察结果一致；然而，即使是较低浓度的 12-KETE 也会激活滋养层蛋白酶。同时，源自牛胎盘的成纤维细胞将花生四烯酸转化为 12-KETE。结论这些观察表明 12-KETE 可作为分娩时胎盘分离的信号。

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12-Oxoeicosatetraenoic acid, a candidate signal for placenta separation, activates matrix metalloproteinase and induces apoptosis in bovine trophoblast cells.

OBJECTIVE 12-oxo-5Z,8Z,10E,14Z-eicosatetraenoic acid (12-KETE), a metabolite of arachidonic acid, is a strong candidate signal for placenta separation following calf discharge at delivery. In the present study, the effects of 12-KETE on bovine trophoblast cells were investigated to determine its function in the placentome at delivery. METHODS Bovine trophoblast cells derived from blastocysts were used. They were cocultured with or without fibroblasts derived from bovine placentome and/or bovine uterine epithelial cells. 12-KETE was added to the culture medium. RESULTS Bovine trophoblast cells contained binucleate cells and strongly expressed caudal type homeobox 2 (CDX-2) genes. Addition of 12-KETE to the trophoblast cell colony without feeder cells or that on a fibroblast monolayer induced rapid exfoliation of the colony. After 12-KETE addition, trophoblast cells emitted strong fluorescence caused by the degradation of dye-quenched collagen, indicating that 12-KETE activated matrix metalloproteinase of the trophoblast cells. Exfoliated cell colonies were stained with YOPRO-1, but not propidium iodide (PI). Moreover, DNA fragmentation and Bcl-2 associated X protein (Bax) gene (apoptosis stimulator) upregulation were observed in exfoliated cells, indicating that 12-KETE induced trophoblast cell apoptosis. These results were consistent with previous in vivo observations; however, even a lower concentration of 12-KETE activated trophoblast protease. Meanwhile, fibroblasts derived from the bovine placentome converted arachidonic acid to 12-KETE. CONCLUSION These observations indicate that 12-KETE may serve as a signal for placenta separation at delivery.

更新日期：2022-06-30

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