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体外成熟培养期间补充白藜芦醇可改善通过灿烂甲酚蓝染色筛选的青春期前山羊卵母细胞的胚胎发育。

Resveratrol supplementation during in vitro maturation improves embryo development of prepubertal goat oocytes selected by brilliant cresyl blue staining.

作者信息

Piras Anna-Rita, Menéndez-Blanco Irene, Soto-Heras Sandra, Catalá Maria-Gracia, Izquierdo Dolors, Bogliolo Luisa, Paramio Maria-Teresa

机构信息

Universitat Autònoma de Barcelona, Departament de Ciència Animal i dels Aliments, 08193 Bellaterra, Barcelona, Spain.

Università degli Studi di Sassari, Dipartimento di Medicina Veterinaria, 07100 Sassari, Italy.

出版信息

J Reprod Dev. 2019 Apr 12;65(2):113-120. doi: 10.1262/jrd.2018-077. Epub 2018 Dec 29.

Abstract

This study aimed to investigate the effect of resveratrol supplementation in maturation medium on the developmental ability and bioenergetic\oxidative status of prepubertal goat oocytes selected by brilliant cresyl blue (BCB). Oocytes collected from slaughterhouse-derived ovaries were selected by 13 µM BCB staining and classified as grown BCB+ and growing BCB- oocytes. All oocytes were matured in vitro in our conventional maturation medium and supplemented with 1 µM (BCB+R and BCB-R) and without (Control groups: BCB+C and BCB-C) resveratrol. After 24 h, IVM-oocytes were fertilized with fresh semen and presumptive zygotes were in vitro cultured for 8 days. Oocytes were assessed for blastocyst development and quality, mitochondrial activity and distribution, and levels of GSH, ROS, and ATP. BCB+R (28.3%) oocytes matured with resveratrol presented significantly higher blastocyst development than BCB+C (13.0%) and BCB- groups (BCB-R: 8.3% and BCB-C: 4.7%). Resveratrol improved blastocyst development of BCB-R oocytes at the same rate as BCB+C oocytes. No differences were observed in blastocyst quality among groups. GSH levels were significantly higher in resveratrol groups (BCB+R: 36554.6; BCB-R: 34946.7 pixels/oocyte) than in control groups (BCB+C: 27624.0; BCB-C: 27655.4 pixels/oocyte). No differences were found in mitochondrial activity, ROS level, and ATP content among the groups. Resveratrol-treated oocytes had a higher proportion of clustered active mitochondria in both BCB groups (BCB+R: 73.07%; BCB-R: 79.16%) than control groups (BCB+C: 19.35%; BCB-C: 40%). In conclusion, resveratrol increased blastocyst production from oocytes of prepubertal goats, particularly in better quality oocytes (BCB+).

摘要

本研究旨在探讨在成熟培养液中添加白藜芦醇对经灿烂甲酚蓝(BCB)筛选的青春期前山羊卵母细胞发育能力及生物能量/氧化状态的影响。从屠宰场采集的卵巢中收集的卵母细胞,通过13µM BCB染色进行筛选,并分为成熟的BCB+和生长中的BCB-卵母细胞。所有卵母细胞在我们常规的成熟培养液中进行体外成熟培养,添加1µM白藜芦醇(BCB+R和BCB-R组)或不添加(对照组:BCB+C和BCB-C组)。24小时后,体外成熟的卵母细胞用新鲜精液受精,假定的受精卵进行8天的体外培养。评估卵母细胞的囊胚发育及质量、线粒体活性和分布以及谷胱甘肽(GSH)、活性氧(ROS)和三磷酸腺苷(ATP)水平。添加白藜芦醇成熟的BCB+R(28.3%)卵母细胞的囊胚发育率显著高于BCB+C(13.0%)组和BCB-组(BCB-R:8.3%,BCB-C:4.7%)。白藜芦醇使BCB-R卵母细胞的囊胚发育改善程度与BCB+C卵母细胞相同。各组间囊胚质量无差异。白藜芦醇组(BCB+R:36554.6;BCB-R:34946.7像素/卵母细胞)的GSH水平显著高于对照组(BCB+C:27624.0;BCB-C:27655.4像素/卵母细胞)。各组间线粒体活性、ROS水平和ATP含量无差异。在两个BCB组中,经白藜芦醇处理的卵母细胞中聚集的活性线粒体比例均高于对照组(BCB+R:73.07%;BCB-R:79.16%)(BCB+C:19.35%;BCB-C:40%)。总之,白藜芦醇提高了青春期前山羊卵母细胞的囊胚产量,尤其是质量较好的卵母细胞(BCB+)。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/512e/6473110/9d2b07bcbe08/jrd-65-113-g001.jpg

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