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在体内产生的绵羊胚胎玻璃化或解冻过程中补充左旋肉碱不会影响胚胎存活率,但会改变肉碱乙酰转移酶(CrAT)和过氧化物还原酶1(PRDX1)的表达。

l-carnitine supplementation during vitrification or warming of in vivo-produced ovine embryos does not affect embryonic survival rates, but alters CrAT and PRDX1 expression.

作者信息

Saraiva Helena F R A, Batista Ribrio I T P, Alfradique Vivian A P, Pinto Pedro H N, Ribeiro Lilian S, Oliveira Clara S, Souza-Fabjan Joanna M G, Camargo Luiz S A, Fonseca Jeferson F, Brandão Felipe Z

机构信息

Universidade Federal Fluminense, Vital Brazil St., 64, 24230-240 Niterói, RJ, Brazil.

Universidade Federal Fluminense, Vital Brazil St., 64, 24230-240 Niterói, RJ, Brazil.

出版信息

Theriogenology. 2018 Jan 1;105:150-157. doi: 10.1016/j.theriogenology.2017.09.022. Epub 2017 Sep 21.

Abstract

l-carnitine is an antioxidant and β-oxidation stimulator substance commonly used to improve metabolic performance of oocytes and embryos in in vitro systems. However, few studies have evaluated its beneficial effects in embryos produced in vivo. This study aimed to evaluate the effect of l-carnitine supplementation into vitrification or warming solutions on the post-warming character of day 6-7 in vivo-produced ovine embryos. l-carnitine (3.72 mM) was added to vitrification (Experiment 1) or warming solutions (Experiment 2). In experiments 1 and 2, the embryos were vitrified using straw and cryo-tip protocols, respectively. In vitro culture (IVC) of warmed embryos was performed for 72 h in order to evaluate survival rates, reactive oxygen species (ROS) levels, total cell number (TCN), number of apoptotic cells, apoptotic index evaluation, and gene expression analysis of carnitine palmitoyltransferase I and 2 (CPT1 and CPT2), carnitine O-acetyltransferase (CrAT), and peroxiredoxin-1 (PRDX1). In experiment 1, survival rate, ROS levels after 24 h of IVC, total cell number at 24 h and 72 h, apoptotic cells and apoptotic index at 72 h of IVC were similar in embryos vitrified in medium supplemented with LC or not. Gene expression analysis showed no differences in CPT1 and CPT2 mRNA relative abundance in embryos of both experiments compared to fresh embryos (FE); however, CrAT was downregulated (p < 0.05) in C1, and PRDX1 was downregulated (p < 0.05) in both the control (C1) and l-carnitine (LC1) groups, compared to FE. Moreover, CrAT and PRDX1 were upregulated (p < 0.05) in C2, and CrAT was downregulated (p < 0.05) in LC2, in relation to FE. Although the short-term LC supplementation at 3.72 mM did not improve survival, and quality parameters of in vivo-produced ovine embryos, it could affect their quality at a molecular level. In conclusion, further investigations with different concentrations of LC and tools are needed for improvement of the efficiency of these strategies.

摘要

左旋肉碱是一种抗氧化剂和β-氧化刺激物质,常用于体外系统中改善卵母细胞和胚胎的代谢性能。然而,很少有研究评估其对体内产生的胚胎的有益作用。本研究旨在评估在玻璃化或复温溶液中添加左旋肉碱对体内产生的第6 - 7天绵羊胚胎复温后特性的影响。将左旋肉碱(3.72 mM)添加到玻璃化溶液(实验1)或复温溶液(实验2)中。在实验1和2中,胚胎分别使用细管和冷冻头方案进行玻璃化。对复温后的胚胎进行72小时的体外培养(IVC),以评估存活率、活性氧(ROS)水平、总细胞数(TCN)、凋亡细胞数、凋亡指数评估以及肉碱棕榈酰转移酶I和2(CPT1和CPT2)、肉碱O-乙酰转移酶(CrAT)和过氧化物酶还原蛋白-1(PRDX1)的基因表达分析。在实验1中,在添加或不添加LC的培养基中玻璃化的胚胎,其存活率、IVC 24小时后的ROS水平、24小时和72小时的总细胞数、IVC 72小时时的凋亡细胞和凋亡指数相似。基因表达分析表明,与新鲜胚胎(FE)相比,两个实验中胚胎的CPT1和CPT2 mRNA相对丰度没有差异;然而,与FE相比,C1组中CrAT下调(p < 0.05),对照组(C1)和左旋肉碱(LC1)组中PRDX1均下调(p < 0.05)。此外,与FE相比,C2组中CrAT和PRDX1上调(p < 0.05),LC2组中CrAT下调(p < 0.05)。虽然3.72 mM的短期LC补充并没有提高体内产生的绵羊胚胎的存活率和质量参数,但它可能在分子水平上影响其质量。总之,需要用不同浓度的LC和方法进行进一步研究,以提高这些策略的效率。

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