Grzesiak Malgorzata, Knapczyk-Stwora Katarzyna, Ciereszko Renata E, Wieciech Iwona, Slomczynska Maria
Department of Endocrinology, Institute of Zoology, Jagiellonian University in Krakow, Krakow, Poland.
Department of Endocrinology, Institute of Zoology, Jagiellonian University in Krakow, Krakow, Poland.
Theriogenology. 2014 Sep 15;82(5):720-33. doi: 10.1016/j.theriogenology.2014.06.005. Epub 2014 Jun 11.
Recently, we have found that flutamide-induced androgen deficiency altered progesterone production in the porcine corpus luteum (CL) during mid- and late pregnancy. Herein, we tested whether flutamide administration subsequently influences androgen and estrogen metabolism in the CL of pregnancy. Pregnant gilts were treated with flutamide between Days 43 and 49 (GD50F), 83 and 89 (GD90F), or 101 and 107 (GD108F) of gestation. Corpora lutea (CLs) were collected from treated and nontreated (control) pigs. The concentrations of androstenedione (A4), testosterone (T), estrone (E1), and estradiol (E2) together with the levels of expression of mRNAs and proteins for cytochrome P450 17α-hydroxylase/c17-20 lyase (CYP17A1), 17β-hydroxysteroid dehydrogenase type 1 (17β-HSD1), cytochrome P450 aromatase (CYP19A1), and 17β-hydroxysteroid dehydrogenase type 7 (17β-HSD7) were measured in the CL of control and flutamide-treated animals. Steroidogenic enzymes were also immunolocalized in luteal tissues. The luteal concentrations of A4 and T were higher in the GD50F (P = 0.006, P = 0.03) and GD108F (P = 0.005, P = 0.035) groups, but lower in the GD90F (P = 0.004, P = 0.014) group. The E1 level was greater only in the GD90F (P = 0.03) and GD108F (P = 0.035) groups, whereas E2 concentration was not affected by flutamide treatment. Increased luteal CYP17A1 mRNA and protein expression was found in the GD50F (P = 0.002, P = 0.03) and GD108F (P = 0.0026, P = 0.03) groups, but reduced in the GD90F (P = 0.002, P = 0.03) group. mRNA of 17β-HSD1 was upregulated in the GD50F (P = 0.0005) group, but downregulated in the GD90F (P = 0.002) and GD108F (P = 0.0005) groups. In contrast, 17β-HSD1 protein expression was higher in the GD50F and GD108F (P = 0.03) groups, but lower in the GD90F (P = 0.03) group. Both CYP19A1 mRNA and protein levels were greater in the GD90F (P = 0.001, P = 0.028) and GD108F (P = 0.005, P = 0.03) groups. Neither 17β-HSD7 mRNA nor protein level were affected by flutamide exposure. Both CYP17A1 and 17β-HSD1 were immunolocalized exclusively in small luteal cells, whereas CYP19A1 and 17β-HSD7 were found in large luteal cells of control and flutamide-treated CLs. Overall, flutamide administration led to the alterations in A4, T, and E1, but not in E2, production in the CL of pregnancy in pigs, probably because of disrupted steroidogenic enzymes expression. These changes suggest that androgens are important modulators of luteal function during pregnancy in pigs.
最近，我们发现氟他胺诱导的雄激素缺乏会改变妊娠中后期猪黄体（CL）中孕酮的产生。在此，我们测试了氟他胺给药是否随后会影响妊娠黄体中雄激素和雌激素的代谢。在妊娠第43至49天（GD50F）、83至89天（GD90F）或101至107天（GD108F）对妊娠母猪进行氟他胺处理。从处理过和未处理（对照）的猪中收集黄体（CLs）。测量对照和氟他胺处理动物的黄体中雄烯二酮（A4）、睾酮（T）、雌酮（E1）和雌二醇（E2）的浓度，以及细胞色素P450 17α-羟化酶/c17-20裂解酶（CYP17A1）、17β-羟类固醇脱氢酶1型（17β-HSD1）、细胞色素P450芳香化酶（CYP19A1）和17β-羟类固醇脱氢酶7型（17β-HSD7）的mRNA和蛋白质表达水平。类固醇生成酶也在黄体组织中进行免疫定位。GD50F组（P = 0.006，P = 0.03）和GD108F组（P = 0.005，P = 0.035）的黄体中A4和T浓度较高，但GD90F组（P = 0.004，P = 0.014）较低。E1水平仅在GD90F组（P = 0.03）和GD108F组（P = 0.035）中较高，而E2浓度不受氟他胺处理的影响。在GD50F组（P = 0.002，P = 0.03）和GD108F组（P = 0.0026，P = 0.03）中发现黄体CYP17A1 mRNA和蛋白质表达增加，但在GD90F组（P = 0.002，P = 0.03）中降低。17β-HSD1的mRNA在GD50F组（P = 0.0005）中上调，但在GD90F组（P = 0.002）和GD108F组（P = 0.0005）中下调。相比之下，17β-HSD1蛋白质表达在GD50F和GD108F组（P = 0.03）中较高，但在GD90F组（P = 0.03）中较低。GD90F组（P = 0.001，P = 0.028）和GD108F组（P = 0.005，P = 0.03）中CYP19A1的mRNA和蛋白质水平均较高。氟他胺暴露对17β-HSD7的mRNA和蛋白质水平均无影响。CYP17A1和17β-HSD1仅在小黄体细胞中进行免疫定位，而CYP19A1和17β-HSD7在对照和氟他胺处理的CLs的大黄体细胞中发现。总体而言，氟他胺给药导致猪妊娠黄体中A4、T和E1的产生发生改变，但E2未受影响，这可能是由于类固醇生成酶表达受到干扰。这些变化表明雄激素是猪妊娠期间黄体功能的重要调节因子。
