湖羊黄体期注射PGF2α对黄体组织形态、PGF2α受体和程序性坏死相关基因表达的影响

doi:10.11843/j.issn.0366-6964.2023.04.014

摘要/Abstract

摘要： 旨在研究不同黄体期注射前列腺激素（PGF2α）对湖羊育成母羊黄体组织形态、PGF2α受体及程序性坏死通路基因表达的影响。本研究选择健康、体况良好、体重相近经同期发情处理的母羊48只，随机均分为6组，分别为黄体前期试验组和对照组、中期试验组和对照组和末期试验组和对照组母羊，试验组在发情周期第6天（黄体前期）、11天（黄体中期）、16天（黄体末期）注射1 mL PGF2α（0.1 mg），对照组在相同时间注射1 mL生理盐水，每次注射后3 h采集黄体组织，用于基因表达检测及HE组织染色检测黄体组织形态变化。结果表明，注射PGF2α后试验羊黄体组织形态发生了变化，出现炎症细胞浸润与细胞凋亡，其中黄体中期退化程度最高。未注射PGF2α的母羊，黄体前期、中期FPA mRNA表达水平显著高于黄体末期（P<0.05）；黄体前期FPB表达水平显著高于黄体末期（P<0.05），与黄体中期无显著差异（P>0.05）；黄体中期TNFR1表达水平显著高于黄体前期和末期（P<0.05）；黄体中期RIPK3表达水平显著高于黄体末期（P<0.05），黄体中、末期与黄体前期差异均不显著（P>0.05）；TNF-α、RIPK1和MLKL表达水平在整个黄体期没有显著变化（P>0.05）。黄体前期注射PGF2α显著降低FPA表达水平（P<0.05），显著提高FPB、RIPK1、MLKL表达量（P<0.05），对TNF-α、TNFR1、RIPK3基因表达无显著影响（P>0.05）；黄体中期注射PGF2α可以显著上调FPB、TNF-α、RIPK1与MLKL表达量（P<0.05），对FPA、TNFR1和RIPK3表达没有显著影响（P>0.05）；黄体末期注射PGF2α可以显著上调RIPK1表达（P<0.05），对FPA、FPB、TNF-α、RIPK3和MLKL表达没有显著影响（P>0.05）。FPA与RIPK3和TNFR1 mRNA表达呈正相关（P<0.05），FPB与MLKL和TNF-α mRNA表达呈正相关（P<0.05）。结果表明，PGF2α与其受体FPA、FPB结合后可能通过TNF-α/TNFR1通路启动程序性坏死，促进黄体组织退化，FPB在介导PGF2α诱导黄体退化和程序性坏死中发挥主要作用，PGF2α在黄体中期诱导黄体退化效果较好。

关键词: 母羊, 前列腺素F2α, 黄体期, 程序性坏死, 前列腺素受体

Abstract: The purpose of this experiment was to study the effects of PGF2α injection at different luteal stages on luteal tissue morphology, expression of PGF2α receptors and programmed necroptosis-associated genes in Hu sheep. Forty eight healthy ewes with similar body weight were used in this study, which were in good condition and have undergone estrus synchronization. They were randomly divided into 6 groups, namely the early-luteal phase experiment group and control group, the mid-luteal phase experiment group and control group, and the late-luteal phase experiment group and control group. The ewes in 3 experimental groups were injected with 1 mL PGF2α (0.1 mg) on day 6 (early-luteal phase), day 11 (mid-luteal phase) and day 16 (late-luteal phase), respectively. The ewes in 3 control groups were injected with 1 mL normal saline at the same time. The corpus luteum tissue was collected at 3 h after each injection for gene expression detection and HE staining for the detection of morphological changes of corpus luteum. The results showed that, after the injection of PGF2α, the morphology of corpus luteum changed, with inflammatory cell infiltration and apoptosis, especially in the mid-luteal phase, with a higher degree of degeneration. For ewes that were not injected with PGF2α, the mRNA expression level of FPA in the corpus luteum in the early-luteal phase was significantly higher in the early-luteal and mid-luteal phases compared to the late-luteal phase (P<0.05); the FPB expression level in the corpus luteum in the early-luteal phase was significantly higher than that in the late-luteal phase (P<0.05), and there was no significant difference from the mid-luteal phase (P>0.05); the expression level of TNFR1 in the mid-luteal phase was significantly higher than that in the early-luteal and late-luteal phases (P<0.05); The expression level of RIPK3 in the mid-luteal phase was significantly higher than that in the late-luteal phase (P<0.05), and both of them was no significant difference to the early-luteal phase (P>0.05); TNF-α, RIPK1 and MLKL expression levels did not significantly change throughout the luteal phase (P>0.05). In the early-luteal phase, injection of PGF2α significantly reduced the expression of FPA (P<0.05), and significantly increased the expression of FPB, RIPK1, MLKL (P<0.05), and had no significant effect on the expression of TNF-α, TNFR1, and RIPK3 (P>0.05).In the mid-luteal phase, injection of PGF2α could significantly up-regulate the expression of FPB, TNF-α, RIPK1 and MLKL (P<0.05), but had no significant effect on the expression of FPA, TNFR1 and RIPK3 (P>0.05); In the late-luteal phase, injection of PGF2α could significantly up-regulate the expression of RIPK1 (P<0.05)), had no effect on the expression of FPA, FPB, TNF-α, RIPK3 and MLKL (P>0.05). The mRNA expression level of FPA was positively correlated with RIPK3 and TNFR1 (P<0.05), and the mRNA expression level of FPB was positively correlated with MLKL and TNF-α (P<0.05). The results show that PGF2α may initiate necroptosis through the TNF-α/TNFR1 pathway after binding to its receptors FPA and FPB, and promote the degeneration of the corpus luteum. FPB plays a major role in mediating PGF2α-induced luteal degeneration and necroptosis, and PGF2α has a better effect on inducing luteal degeneration in the mid-luteal phase.

Key words: ewe, prostaglandin F2α, luteal phase, necroptosis, prostaglandin F receptor

中图分类号:

S826.3

李宇, 段春辉, 宋志攀, 岳思聪, 王媛, 张英杰, 刘月琴. 湖羊黄体期注射PGF2α对黄体组织形态、PGF2α受体和程序性坏死相关基因表达的影响[J]. 畜牧兽医学报, 2023, 54(4): 1500-1510.

LI Yu, DUAN Chunhui, SONG Zhipan, YUE Sicong, WANG Yuan, ZHANG Yingjie, LIU Yueqin. Effects of PGF2α on Luteal Tissue Morphology, Expression of PGF2α Receptors and Necroptosis-associated Genes During Luteal Phase in Hu Sheep[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(4): 1500-1510.

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