Seto Nickie L, Bogan Randy L
School of Animal and Comparative Biomedical Sciences, College of Agriculture and Life Sciences, University of Arizona, Tucson, Arizona.
School of Animal and Comparative Biomedical Sciences, College of Agriculture and Life Sciences, University of Arizona, Tucson, Arizona
Biol Reprod. 2015 May;92(5):128. doi: 10.1095/biolreprod.114.124941. Epub 2015 Apr 16.
In nonprimate species, it has been well established that prostaglandin F2 alpha (PGF2alpha) initiates luteolysis. Changes in intracellular cholesterol concentrations caused by modulation of cholesterol uptake and efflux may mediate PGF2alpha-induced luteolysis. These changes in cholesterol efflux and uptake are controlled, in part, by the liver x receptors (LXR) alpha (NR1H3) and beta (NR1H2), nuclear receptors that increase expression of genes necessary for cholesterol efflux or limiting cholesterol uptake. Therefore, we hypothesized that PGF2alpha reduces expression of cholesterol uptake and increases expression of cholesterol efflux genes, mediated in part by enhanced LXR activity. To test this hypothesis, an induced luteolysis model was used whereby ewes were treated during their midluteal phase with saline or PGF2alpha and corpora lutea (CL) collected 12, 24, or 48 h later for determination of mRNA and protein concentrations by quantitative real-time PCR and Western blot analysis, respectively. As a complementary approach, CL undergoing spontaneous luteolysis were compared to midluteal phase CL. The lipoprotein receptors responsible for cholesterol uptake were significantly decreased in both luteolysis models. Expression of the LXR target gene ATP binding cassette subfamily A1 (ABCA1), an important mediator of cholesterol efflux, was significantly increased in both experimental models. Chromatin immunoprecipitation confirmed that PGF2alpha treatment resulted in enhanced NR1H3 and NR1H2 binding to the ABCA1 promoter. Qualitative changes in lipid droplet distribution were also observed following PGF2alpha treatment. These data support the hypothesis that reduced cholesterol uptake and increased efflux mediate luteolysis in sheep, which is partially controlled by PGF2alpha stimulation of LXR activity.
在非灵长类物种中,前列腺素F2α(PGF2α)引发黄体溶解已得到充分证实。由胆固醇摄取和流出调节引起的细胞内胆固醇浓度变化可能介导PGF2α诱导的黄体溶解。这些胆固醇流出和摄取的变化部分受肝脏X受体(LXR)α(NR1H3)和β(NR1H2)控制,这两种核受体可增加胆固醇流出或限制胆固醇摄取所需基因的表达。因此,我们假设PGF2α会降低胆固醇摄取基因的表达并增加胆固醇流出基因的表达,部分是由增强的LXR活性介导的。为了验证这一假设,我们使用了诱导黄体溶解模型,即在母羊的黄体中期用生理盐水或PGF2α进行处理,然后在12、24或48小时后收集黄体(CL),分别通过定量实时PCR和蛋白质印迹分析来测定mRNA和蛋白质浓度。作为一种补充方法,将经历自发黄体溶解的CL与黄体中期的CL进行比较。在两种黄体溶解模型中,负责胆固醇摄取的脂蛋白受体均显著减少。LXR靶基因ATP结合盒亚家族A1(ABCA1)是胆固醇流出的重要介质,其表达在两种实验模型中均显著增加。染色质免疫沉淀证实,PGF2α处理导致NR1H3和NR1H2与ABCA1启动子的结合增强。PGF2α处理后还观察到脂滴分布的定性变化。这些数据支持了以下假设:胆固醇摄取减少和流出增加介导了绵羊的黄体溶解,这部分受PGF2α对LXR活性的刺激控制。