畜牧兽医学报 ›› 2021, Vol. 52 ›› Issue (6): 1582-1593.doi: 10.11843/j.issn.0366-6964.2021.06.013

• 生物技术与繁殖 • 上一篇    下一篇

前列腺素D2与F对绵羊黄体退化的影响及其机理研究

杨恒1*, 邵焱焱2, 赵宗胜2*, 朱梦婷2,3, 南颖2, 王明远2, 方晨辉2, 吴培2, 谢梦婷2, 江白慧2   

  1. 1. 西南大学动物医学院, 重庆 402460;
    2. 石河子大学动物科技学院, 石河子 832000;
    3. 新疆农垦科学院 省部共建绵羊遗传改良与健康养殖国家重点实验室, 石河子 832000
  • 收稿日期:2021-03-26 出版日期:2021-06-23 发布日期:2021-06-22
  • 通讯作者: 杨恒,主要从事动物遗传育种与繁殖研究,E-mail:yh20183007@swu.edu.cn;赵宗胜,要从事动物遗传育种与繁殖研究,E-mail:zhaozongsh@shzu.edu.cn
  • 作者简介:杨恒(1986-),男,重庆荣昌人,博士,讲师,主要从事动物遗传育种与繁殖研究,E-mail:yh20183007@swu.edu.cn;邵焱焱(1998-),女,新疆北屯人,硕士生,主要从事动物遗传育种与繁殖研究,E-mail:2843437982@qq.com。杨恒和邵焱焱为同等贡献作者
  • 基金资助:
    国家自然科学基金(31802065);重庆市自然科学基金项目(cstc2020jcyj-msxmX0427);西南大学博士启动基金(SWU119013);省部共建绵羊遗传改良与健康养殖国家重点实验室(MYSKLKF201902)

Study on the Effect and Mechanism of Prostaglandin D2 and F on the Corpus Luteum Regression in Sheep

YANG Heng1*, SHAO Yanyan2, ZHAO Zongsheng2*, ZHU Mengting2,3, NAN Ying2, WANG Mingyuan2, FANG Chenhui2, WU Pei2, XIE Mengting2, JIANG Baihui2   

  1. 1. College of Veterinary Medicine, Southwest University, Chongqing 402460, China;
    2. College of Animal Science and Technology, Shihezi University, Shihezi 832000, China;
    3. State Key Laboratory of Sheep Genetic Improvement and Healthy Breeding, Xinjiang Academy of Agricultural and Reclamation Sciences, Shihezi 832000, China
  • Received:2021-03-26 Online:2021-06-23 Published:2021-06-22

摘要: 旨在研究前列腺素(prostaglandins,PGs)D2与F对绵羊黄体(corpus luteum,CL)组织形态、生殖激素及其关键基因与受体表达的影响,并解析其在黄体退化中的相互关系及机理,为保证母畜连续性繁育提供新的理论依据。将16只哈萨克绵羊随机分成4组,在发情周期的黄体期分别子宫肌内注射PGD2、PGF、PGD2+PGF及等量生理盐水(对照组),采用HE染色结合物理拍照对比处理前后黄体组织形态变化,ELISA法检测外周血清中P4、E2、PGD2和PGF浓度变化;并利用qRT-PCR和Western blot检测关键合成酶基因HPGDS、PGFS及其受体DP1、CRTH2、FP的mRNA和蛋白表达水平。结果显示,与对照组相比,发现PGD2+PGF组中黄体退化效果最明显,随后依次是PGF组明显大于PGD2组。ELISA结果显示,随着处理后时间的推移,不同试验处理组中,P4浓度均呈显著下降趋势(P<0.05),其中在PGD2+PGF组中该变化趋势最显著(P<0.05);但E2、PGD2和PGF浓度均呈现不同差异性变化,其中,PGD2+PGF组中PGD2和PGF浓度呈显著下降(P<0.05),PGF组中E2浓度呈显著升高(P<0.05)、PGD2浓度呈显著下降(P<0.05),PGD2组中E2浓度呈显著下降趋势(P<0.05)、PGD2浓度呈显著升高趋势(P<0.05)。qRT-PCR和Western blot结果显示,与对照组相比,PGD2+PGF2α组中HPGDS mRNA和蛋白表达量显著下调(P<0.05),PGFS、CRTH2及FP mRNA和蛋白表达量显著上调(P<0.05);PGF2α组中HPGDS mRNA和蛋白表达量呈显著下调(P<0.05),其它基因mRNA和蛋白表达量呈显著上调(P<0.05);PGD2组中HPGDS、DP1、PGFS及FP mRNA和蛋白表达量呈显著上调(P<0.05)。同时,在不同受体基因表达量检测时,发现PGD2组中DP1受体表达量显著高于CRTH2受体(P<0.05),而PGF组中CRTH2表达量则显著高于DP1(P<0.05)。综上,PGD2无论单独使用还是结合PGF使用,均能够促进CL的退化,尤其是二者结合时有明显的协同促溶效应,其作用机制可能与其体内激素水平、关键合成酶及受体类型的表达有关,这为全面认识哺乳动物CL退化的调控机制奠定了基础,也为进一步优化高效繁殖技术(尤其是PGs方案)提供了新的思路。

关键词: 绵羊, 黄体退化, PGD2, PGF

Abstract: The objective of this study is to investigate the effects of prostaglandins (PGs) D2 and F on the corpus luteum tissue morphology, reproductive hormones and the expression of key genes and receptors in female sheep. Meanwhile, the interrelationship and mechanism of the above factors in the degradation of the corpus luteum are analyzed, which can provide a new theoretical basis for ensuring the continuous breeding of female animals. Sixteen Kazakh sheep were randomly divided into 4 groups, and were injected intramuscularly in uterus with PGD2, PGF, PGD2+PGF and normal saline (control group) during the luteal phase of the estrus cycle, respectively. HE staining combined with physical photographs were used to compare the morphological changes of the corpus luteum before and after the treatment. ELISA method was used to detect the changes in the concentration of P4, E2, PGD2 and PGFin the peripheral serum; qRT-PCR and Western blot were used to detect the key synthase genes HPGDS, PGFS and their receptors (DP1, CRTH2, FP) mRNA and protein expression levels. Results showed that, compared with the control group, the corpus luteum degeneration effect was the most obvious in the PGD2+PGF group, followed by the PGF group and PGD2 group. ELISA results showed that with the passage of time after treatment, P4 concentration in different experimental treatment groups showed a significant downward trend (P<0.05), and the change trend was the most significant in the PGD2+PGF group (P<0.05). However, the concentrations of E2, PGD2 and PGF showed different differences in different groups. Among them, the PGD2 and PGF concentrations in the PGD2+PGF group were significantly decreased (P<0.05); the E2 concentration was significantly increased (P<0.05), and the PGD2 concentration was significantly decreased (P<0.05) in the PGF group; the E2 concentration was significantly decreased (P<0.05), and the PGD2 concentration was significantly increased (P<0.05) in the PGD2 group. Results of qRT-PCR and Western blot showed that, compared with the control group, HPGDS mRNA and protein expression were significantly down-regulated (P<0.05), but PGFS, CRTH2 and FP mRNA and protein expression were significantly up-regulated (P<0.05) in PGD2+PGF group; In the PGFgroup, HPGDS mRNA and protein expression were significantly down-regulated (P<0.05), and the mRNA and protein expression levels of other genes were significantly up-regulated (P<0.05); In the PGD2 group, the expressions of HPGDS, DP1, PGFS and FP mRNA and protein were significantly up-regulated (P<0.05). Meanwhile, in the detection of gene expression of different receptors, it was found that the expression of DP1 receptor was significantly higher than that of the CRTH2 receptor in the PGD2 group (P<0.05), while the expression of CRTH2 was significantly higher than that of DP1 in the PGF group (P<0.05). In summary, the above results showed that whether PGD2 was used alone or in combination with PGF, it could promote the degradation of CL, especially when the two were combined, which had an obvious synergistic effect to promote dissolution of CL. This mechanism may be related to the hormones level, the expression of key synthetases and receptor types in vivo, which lays a foundation for a comprehensive understanding of the regulatory mechanism of CL degradation in mammals, and provides a new idea for further optimization of efficient reproductive technology (especially PGs scheme).

Key words: sheep, corpus luteum regression, PGD2, PGF

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