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补充白藜芦醇可促进体外生产的牛胚胎玻璃化冷冻及解冻后较低氧化代谢的恢复。

Resveratrol supplementation promotes recovery of lower oxidative metabolism after vitrification and warming of in vitro-produced bovine embryos.

作者信息

Madrid Gaviria Stephania, Morado Sergio A, López Herrera Albeiro, Restrepo Betancur Giovanni, Urrego Álvarez Rodrigo A, Echeverri Zuluaga Julián, Cética Pablo D

机构信息

Grupo de investigación en Biodiversidad y Genética Molecular (BIOGEM), Departamento de Producción Animal, Universidad Nacional de Colombia, Sede Medellín, Carrera 65 No. 59A-110, Código Postal 050034, Colombia.

Universidad de Buenos Aires, Facultad de Ciencias Veterinarias, Instituto de Investigación y Tecnología en Reproducción Animal, Chorroarin 280, Código Postal C1427CWO, Buenos Aires, Argentina.

出版信息

Reprod Fertil Dev. 2019 Mar;31(3):521-528. doi: 10.1071/RD18216.

Abstract

Although vitrification is the current method of choice for oocyte and embryo cryopreservation, it may have detrimental effects on reduction-oxidation status and mitochondrial activity. The aim of this study was to evaluate the effect of supplementing invitro culture (IVC) media and/or vitrification solutions with the antioxidant resveratrol on active mitochondria, mitochondrial superoxide production and lipid peroxidation. Abattoir-derived oocytes were matured and fertilised invitro using standard procedures. Following IVF (21h later), zygotes were cultured in IVC medium supplemented with 0 or 0.5µM resveratrol. On Day 7, blastocysts were vitrified using the Cryotech Vitrification Kit (Cryo Tech Laboratory) with or without 0.5µM resveratrol. After warming, active mitochondria, mitochondrial superoxide production and lipid peroxidation were evaluated using Mito Tracker Green FM, MitoSOX Red and BODIPY581/591 C11 staining respectively. The vitrification-warming process significantly increased active mitochondria and mitochondrial superoxide production in bovine embryos (P<0.05, ANOVA). The addition of 0.5µM resveratrol to the IVC medium or vitrification solutions significantly attenuated the increase in active mitochondria (P<0.05), but not in mitochondrial superoxide production, whereas embryos cultured and vitrified with resveratrol showed the highest values for both parameters (P<0.05). Regarding lipid peroxidation, no significant differences were detected between treatments. In conclusion, resveratrol supplementation of IVC medium or vitrification solutions contributes to recovery of an embryo's 'quieter' state (i.e. lower oxidative metabolism) after vitrification. However, supplementation of both solutions with resveratrol seemed to have a pro-oxidant effect.

摘要

尽管玻璃化是目前卵母细胞和胚胎冷冻保存的首选方法,但它可能对氧化还原状态和线粒体活性产生不利影响。本研究的目的是评估在体外培养(IVC)培养基和/或玻璃化溶液中添加抗氧化剂白藜芦醇对活性线粒体、线粒体超氧化物产生和脂质过氧化的影响。使用标准程序将屠宰场来源的卵母细胞进行体外成熟和受精。体外受精后(21小时后),将受精卵在添加0或0.5μM白藜芦醇的IVC培养基中培养。在第7天,使用Cryotech玻璃化试剂盒(Cryo Tech实验室)对囊胚进行玻璃化,添加或不添加0.5μM白藜芦醇。解冻后,分别使用Mito Tracker Green FM、MitoSOX Red和BODIPY581/591 C11染色评估活性线粒体、线粒体超氧化物产生和脂质过氧化。玻璃化解冻过程显著增加了牛胚胎中的活性线粒体和线粒体超氧化物产生(P<0.05,方差分析)。在IVC培养基或玻璃化溶液中添加0.5μM白藜芦醇可显著减弱活性线粒体的增加(P<0.05),但对线粒体超氧化物产生没有影响,而用白藜芦醇培养和玻璃化的胚胎在这两个参数上的值最高(P<0.05)。关于脂质过氧化,各处理之间未检测到显著差异。总之,在IVC培养基或玻璃化溶液中添加白藜芦醇有助于恢复胚胎在玻璃化后的“更安静”状态(即较低的氧化代谢)。然而,在两种溶液中都添加白藜芦醇似乎具有促氧化作用。

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