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瘦素补充对牛体外卵母细胞成熟和胚胎培养中胚胎发育和基因表达模式的影响。

Effect of leptin supplementation during in vitro oocyte maturation and embryo culture on bovine embryo development and gene expression patterns.

机构信息

Departamento de Fisiología (Fisiología Animal), Facultad de Veterinaria, UCM, Madrid 28040, Spain.

出版信息

Theriogenology. 2011 Mar 15;75(5):887-96. doi: 10.1016/j.theriogenology.2010.10.031. Epub 2010 Dec 31.

Abstract

Leptin is a metabolic hormone related to body condition and nutritional status that influences fertility in assisted reproductive technologies modulating oocyte and embryo quality. The aim of the present study was to establish the effect of various leptin concentrations (0, 10, 100 ng/mL) during in vitro oocyte maturation (IVM) and in vitro embryo culture (IVC) on bovine embryo development and quality in terms of gene expression. The relative mRNA abundance of the genes encoding solute carrier family 2 (facilitated glucose transporter) member 1 (SLC2A1), Bcl-2-associated X protein (BAX), placenta-specific 8 (PLAC8), aldo-keto reductase family 1 member B1 (AKR1B1) and leptin receptor (LEPR) were determined on Day 7 blastocysts by qRT-PCR. Cleavage rate (P < 0.005) and blastocyst yield (P = 0.05) was significantly lower when cumulus-oocyte complexes (COCs) were matured with 100 ng/mL leptin compared to 0 or 10 ng/mL leptin. No significant effect of different concentrations of leptin added during IVC on blastocyst yield was observed. The presence of 100 ng/mL leptin in both IVM and IVC further decreased cleavage rate (P < 0.005) and blastocyst yield compared to the control group without leptin (P = 0.05) and those supplemented with 10 ng/mL leptin or FCS (P < 0.005). There was no evidence of any leptin-induced difference in the relative transcript abundance of SLC2A1, BAX and PLAC8 genes in Day 7 blastocysts. Expression of AKR1B1 was significantly lower in blastocysts from COCs matured with 100 ng/mL leptin compared to those matured with 0 or 10 ng/mL leptin (P < 0.005). LEPR expression was up regulated when leptin concentration was increased from 0 ng/mL during IVM to 10 ng/mL during IVC, but it was down-regulated in the opposite situation (P < 0.005). In conclusion, high leptin concentrations possibly related to obesity seem to be more detrimental rather than the absence of this hormone for preimplantation embryo survival; this effect is independent of LEPR gene expression and it does not influence expression of SLC2A1, BAX and PLAC8 genes in Day 7 blastocysts.

摘要

瘦素是一种与身体状况和营养状况有关的代谢激素,它通过调节卵母细胞和胚胎质量来影响辅助生殖技术的生育能力。本研究的目的是确定在体外卵母细胞成熟(IVM)和体外胚胎培养(IVC)过程中,不同浓度(0、10、100ng/ml)的瘦素对牛胚胎发育和基因表达的影响。qRT-PCR 法测定第 7 天囊胚中编码溶质载体家族 2(促进葡萄糖转运体)成员 1(SLC2A1)、Bcl-2 相关 X 蛋白(BAX)、胎盘特异性 8(PLAC8)、醛酮还原酶家族 1 成员 B1(AKR1B1)和瘦素受体(LEPR)的相对 mRNA 丰度。与 0 或 10ng/ml 瘦素相比,当用 100ng/ml 瘦素成熟卵丘-卵母细胞复合物(COCs)时,卵裂率(P<0.005)和囊胚产量(P=0.05)显著降低。在 IVC 过程中添加不同浓度的瘦素对囊胚产量无显著影响。与不含瘦素的对照组(P=0.05)和添加 10ng/ml 瘦素或 FCS 的对照组(P<0.005)相比,IVM 和 IVC 中均添加 100ng/ml 瘦素进一步降低了卵裂率(P<0.005)和囊胚产量。在第 7 天囊胚中,没有证据表明 SLC2A1、BAX 和 PLAC8 基因的相对转录丰度存在任何由瘦素引起的差异。与用 0 或 10ng/ml 瘦素成熟的 COCs 相比,用 100ng/ml 瘦素成熟的 COCs 的 AKR1B1 表达显著降低(P<0.005)。当 IVM 期间瘦素浓度从 0ng/ml 增加到 IVC 期间的 10ng/ml 时,LEPR 表达上调,但在相反情况下下调(P<0.005)。总之,高浓度的瘦素(可能与肥胖有关)似乎对胚胎植入前的存活更为不利,而不是缺乏这种激素;这种影响与 LEPR 基因表达无关,也不影响第 7 天囊胚中 SLC2A1、BAX 和 PLAC8 基因的表达。

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