Rak-Mardyła Agnieszka, Durak Martyna, Lucja Gregoraszczuk Ewa
Reprod Biol Endocrinol. 2013 May 17;11:45. doi: 10.1186/1477-7827-11-45.
Resistin was first reported to be an adipocyte-specific hormone, but recent studies have indicated a connection between resistin and reproductive function. However, it is not yet known if resistin is expressed by the ovary and if it can affect steroidogenesis in ovarian follicles from prepubertal pigs.
In this study, using real time PCR, immunoblotting, and ELISA, we quantified resistin expression and concentration in maturing ovarian follicles (small, 3-4 mm; medium, 4-5 mm; large, 6-7 mm) collected from prepubertal pigs. In addition, the dose-responsive effects of recombinant human resistin (0.1, 1, 10, and 100 ng/ml) on steroid hormone (i.e., progesterone [P4], androstendione [A4], testosterone [T], and estradiol [E2]) secretion in culture medium and steroidogenic enzyme (i.e., CYP11A1, 3betaHSD, CYP17A1, 17betaHSD, and CYP19A1) expression in ovarian follicles were determined.
We observed that resistin gene and protein expression increased significantly (P < 0.05) during follicular growth, with large follicles expressing the highest level of this adipokine. Recombinant resistin also increased P4, A4, and T secretion by up-regulating the steady state levels of CYP11A1, 3betaHSD, CYP17A1, and 17betaHSD. Recombinant resistin had no effects on E2 secretion and CYP19A1 expression in ovarian follicles.
Our results show resistin expression in ovarian follicles from prepubertal pigs for the first time. We also show that recombinant resistin stimulates steroidogenesis in ovarian follicles by increasing the expression of CYP11A1, 3betaHSD, CYP17A1, and 17betaHSD. The presence of resistin in the porcine ovary and its direct effects on steroidogenesis suggest that resistin is a new regulator of ovary function in prepubertal animals.
抵抗素最初被报道为一种脂肪细胞特异性激素,但最近的研究表明抵抗素与生殖功能之间存在联系。然而,尚不清楚抵抗素是否由卵巢表达,以及它是否会影响青春期前猪卵巢卵泡中的类固醇生成。
在本研究中,我们使用实时PCR、免疫印迹和ELISA技术,对从青春期前猪采集的成熟卵巢卵泡(小卵泡,3 - 4毫米;中卵泡,4 - 5毫米;大卵泡,6 - 7毫米)中的抵抗素表达和浓度进行了定量分析。此外,还测定了重组人抵抗素(0.1、1、10和100纳克/毫升)对培养基中类固醇激素(即孕酮[P4]、雄烯二酮[A4]、睾酮[T]和雌二醇[E2])分泌以及卵巢卵泡中类固醇生成酶(即CYP11A1、3βHSD、CYP17A1、17βHSD和CYP19A1)表达的剂量反应效应。
我们观察到,在卵泡生长过程中,抵抗素基因和蛋白表达显著增加(P < 0.05),大卵泡中这种脂肪因子的表达水平最高。重组抵抗素还通过上调CYP11A1、3βHSD、CYP17A1和17βHSD的稳态水平,增加了P4、A4和T的分泌。重组抵抗素对卵巢卵泡中的E2分泌和CYP19A1表达没有影响。
我们的结果首次显示了青春期前猪卵巢卵泡中抵抗素的表达。我们还表明,重组抵抗素通过增加CYP11A1、3βHSD、CYP17A1和17βHSD的表达来刺激卵巢卵泡中的类固醇生成。猪卵巢中抵抗素的存在及其对类固醇生成的直接影响表明,抵抗素是青春期前动物卵巢功能的一种新调节因子。