Laboratory for Veterinary Physiology, Department of Veterinary Sciences, University of Antwerp, Universiteitsplein 1, B-2610 Wilrijk, Belgium.
Hum Reprod. 2010 Mar;25(3):768-78. doi: 10.1093/humrep/dep420. Epub 2009 Dec 10.
Obesity is associated with female reproductive abnormalities. Hyperlipidaemia might alter the embryonic micro-environment and potentially result in reduced fertility. We aimed to induce hyperlipidaemia nutritionally and investigate the consequences of hyperlipidaemic culture conditions on bovine in vitro embryo development, embryo quality and gene expression patterns.
Bovine zygotes (n = 1545) were cultured in synthetic oviductal fluid medium supplemented with serum from heifers (n = 3), each fed three successive dietary treatments: (i) control serum, following a hay-based diet, (ii) hyperlipidaemic serum, following a carbohydrate and protected palm-oil-rich diet (FatCh) or (iii) hyperlipidaemic serum, following a protected palm-oil-rich diet (Fat). Blastocysts were evaluated for development, cell count, picnotic and mitotic indexes and cryotolerance. Selected mRNA transcripts were measured by quantitative RT-PCR.
FatCh and Fat diets approximately doubled the total cholesterol concentrations, compared with controls (167.1 +/- 11.9, 150.0 +/- 12.8 versus 83.4 +/- 13.7 mg/dl, respectively, P < 0.05), and fatty acid concentrations (8146.60 +/- 214.61, 6935.56 +/- 1081.04 versus 3944.0 +/- 425.07 micromol/l, respectively, P < 0.05). Supplementation of culture media with FatCh and Fat serum significantly reduced blastocyst rates, compared with controls (27.8, 23.4% versus 36.2%, respectively, P < 0.01), total cell number (103.3 +/- 30.1, 95.6 +/- 28.2 versus 146.9 +/- 34.2, respectively, P < 0.01), mitotic index (1.3 +/- 1.1, 1.7 +/- 2.4 versus 3.6 +/- 2.2%, respectively, P < 0.01) and hatching rates after vitrification (20.4, 13.8 versus 35.7%, respectively, P = 0.03). Embryos in FatCh and Fat groups exhibited significantly higher mRNA levels for genes related to apoptosis and metabolism, compared with controls.
This combined in vivo and in vitro model indicates that the exposure of preimplantation embryos to hyperlipidaemic conditions may result in reduced embryo quality and developmental potential, possibly resulting in poorer fertility.
肥胖与女性生殖异常有关。高脂血症可能改变胚胎的微环境，并可能导致生育能力下降。我们的目的是通过营养诱导高脂血症，并研究高脂血症培养条件对牛体外胚胎发育、胚胎质量和基因表达模式的影响。
用含有小牛血清的合成输卵管液培养基培养牛受精卵（n=1545），小牛分别连续接受三种不同的饮食处理：（i）基础饮食的对照血清，（ii）富含碳水化合物和保护型棕榈油的高脂血症血清（FatCh），或（iii）富含保护型棕榈油的高脂血症血清（Fat）。对囊胚进行发育、细胞计数、固缩和有丝分裂指数以及冷冻耐受性评估。通过定量 RT-PCR 测量选定的 mRNA 转录本。
与对照组相比（167.1±11.9、150.0±12.8 与 83.4±13.7mg/dl，分别，P<0.05），FatCh 和 Fat 饮食使总胆固醇浓度增加了近两倍，FatCh 和 Fat 血清分别使脂肪酸浓度增加了 77.7%（8146.60±214.61、6935.56±1081.04 与 3944.0±425.07μmol/l，分别，P<0.05）。与对照组相比，培养基中添加 FatCh 和 Fat 血清显著降低了囊胚率（分别为 27.8%、23.4%与 36.2%，P<0.01）、总细胞数（分别为 103.3±30.1、95.6±28.2 与 146.9±34.2，分别，P<0.01）、有丝分裂指数（分别为 1.3±1.1、1.7±2.4 与 3.6±2.2%，分别，P<0.01）和玻璃化后孵化率（分别为 20.4%、13.8%与 35.7%，分别，P=0.03）。与对照组相比，FatCh 和 Fat 组胚胎中与凋亡和代谢相关的基因的 mRNA 水平显著升高。
本体内外联合模型表明，胚胎暴露于高脂血症条件下可能导致胚胎质量和发育潜力下降，从而导致生育力下降。
