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猪卵巢卵泡中抵抗素与过氧化物酶体增殖物激活受体γ的体外相互作用。

In vitro interaction between resistin and peroxisome proliferator-activated receptor γ in porcine ovarian follicles.

作者信息

Rak-Mardyła Agnieszka, Drwal Eliza

机构信息

Department of Physiology and Toxicology of Reproduction, Institute of Zoology, Jagiellonian University in Krakow, Gronostajowa 9, 30-387 Krakow, Poland.

出版信息

Reprod Fertil Dev. 2016 Mar;28(3):357-68. doi: 10.1071/RD14053.

Abstract

In the present study, using real-time polymerase chain reaction and immunoblotting methods, we quantified the expression of peroxisome proliferator-activated receptor (PPAR) γ, PPARα and PPARβ in different sized ovarian follicles (small (SF), medium (MF) and large (LF) follicles) in prepubertal and adult pigs. In prepubertal pigs, PPARγ and PPARα expression was highest in LF; however, PPARβ expression did not differ among SF, MF and LF. In mature pigs, only protein expression of PPARγ and PPARα increased during ovarian follicle development. Following identification of very high levels of PPARγ expression in LF in prepubertal and adult pigs, using in vitro culture of ovarian follicles, we determined the effect of resistin at 0.1, 1 and 10ngmL(-1) on PPARγ mRNA and protein expression and the effect of rosiglitazone at 25 and 50µM (a PPARγ agonist) on resistin mRNA and protein expression. Resistin increased PPARγ expression in ovarian follicles in both prepubertal and adult pigs, whereas rosiglitazone had an inhibitory effect on resistin expression. The role of PPARγ in regulating the effects of resistin on ovarian steroidogenesis was investigated using GW9662 (a PPARγ antagonist at dose of 1μM). In these studies, GW9662 reversed the effect of resistin on steroid hormone secretion. The data suggest that there is local cooperation between resistin and PPARγ expression in the porcine ovary. Resistin significantly increased the expression of PPARγ, whereas PPARγ decreased resistin expression; thus, PPARγ is a new key regulator of resistin expression and function.

摘要

在本研究中,我们使用实时聚合酶链反应和免疫印迹法,对青春期前和成年猪不同大小卵巢卵泡(小卵泡(SF)、中卵泡(MF)和大卵泡(LF))中过氧化物酶体增殖物激活受体(PPAR)γ、PPARα和PPARβ的表达进行了定量分析。在青春期前的猪中,PPARγ和PPARα的表达在大卵泡中最高;然而,PPARβ的表达在小卵泡、中卵泡和大卵泡之间没有差异。在成年猪中,只有PPARγ和PPARα的蛋白表达在卵巢卵泡发育过程中增加。在确定青春期前和成年猪的大卵泡中PPARγ表达水平非常高之后,我们利用卵巢卵泡的体外培养,测定了0.1、1和10 ng mL(-1)的抵抗素对PPARγ mRNA和蛋白表达的影响,以及25和50 μM(一种PPARγ激动剂)的罗格列酮对抵抗素mRNA和蛋白表达的影响。抵抗素增加了青春期前和成年猪卵巢卵泡中PPARγ的表达,而罗格列酮对抵抗素的表达有抑制作用。使用GW9662(一种剂量为1 μM的PPARγ拮抗剂)研究了PPARγ在调节抵抗素对卵巢类固醇生成作用中的作用。在这些研究中,GW9662逆转了抵抗素对类固醇激素分泌的影响。数据表明,猪卵巢中抵抗素和PPARγ的表达之间存在局部协同作用。抵抗素显著增加了PPARγ的表达,而PPARγ降低了抵抗素的表达;因此,PPARγ是抵抗素表达和功能的一个新的关键调节因子。

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