INRA, UMR85 Physiologie de la Reproduction et des Comportements, F-37380 Nouzilly, France.
Prostaglandins Other Lipid Mediat. 2010 Sep;93(1-2):30-6. doi: 10.1016/j.prostaglandins.2010.06.002. Epub 2010 Jul 1.
The objectives of the present study were to evaluate the effect of conjugated linoleic acid (CLA t10, c12, C18:2), linolenic acid (C18:3) and docosahexaenoic acid (DHA, C22:6) supplementation on in vitro bovine embryo development, embryo survival after cryopreservation, gene expression and AMPKalpha phosphorylation. Control groups with modified synthetic oviduct fluid (mSOF)+/-100microM beta-mercaptoethanol (beta-ME) were performed. The effects of co-culture with bovine oviduct epithelial cell (Boec) monolayers, serum supplementation and embryo development in the ewe oviduct, on gene expression were also examined. Experiments 1 and 2: a lower d 7 embryo survival was found with 100microM C22:6 and 100microM C18:2 supplementation compared to 1microM C22:6 and 100microM beta-ME supplementation (P<0.05). C18:3 supplementation had no effect on d 7 embryo survival, but 100microM C18:3 increased d 8 embryo survival compared to 100microM beta-ME supplementation (P<0.05). Experiments 3 and 4: stearoyl-CoA desaturase 1 (SCD1) and sterol regulatory element-binding transcription factor 1 (SREBP1) mRNA decreased after 10microM C22:6 supplementation compared to all other supplementations (P<0.05). A lower fatty acid desaturase 2 (FADS2) transcript level was found with 100microM C18:2, 10microM C22:6 and 10microM C18:3 supplementations compared to groups without fatty acid supplementation (P<0.05). Acetyl-CoA-carboxylase (ACC), fatty acid synthase (FAS), adipose differentiation-related protein (ADRP), acyl-CoA synthetase long-chain family member 1 (ACSL1), diacylglycerol O-acyltransferase 1 (DGAT1), carnitin palmitoyltransferase-II (CPT-II) mRNAs expression and AMPKalpha phosphorylation were not modified with PUFA supplementation. Experiment 5: SCD1 and FAS mRNA decrease in Boec group compared to serum supplementation, as SCD1 mRNA in ewe oviduct group (P<0.05). In conclusion, this study showed that a PUFA supplementation with C18:2, C18:3 or C22:6 in bovine culture development for 6 days and co-culture with Boec down-regulate mRNA expression of proteins involved in lipid metabolism in d 7-8 embryo (SCD1 and FADS2 desaturases), probably through SREBP1 mRNA regulation after 10microM C22:6 supplementation, indicating a modification of saturated/unsaturated fatty acid balance in bovine blastocyst.
本研究的目的是评估共轭亚油酸（CLA t10、c12、C18:2）、亚麻酸（C18:3）和二十二碳六烯酸（DHA、C22:6）补充对体外牛胚胎发育、冷冻保存后胚胎存活率、基因表达和 AMPKalpha 磷酸化的影响。使用改良的合成输卵管液（mSOF）+/-100μM β-巯基乙醇（β-ME）进行对照。还研究了与牛输卵管上皮细胞（Boec）单层共培养、血清补充和在母羊输卵管中胚胎发育对基因表达的影响。实验 1 和 2：与 1μM C22:6 和 100μM β-ME 补充相比，100μM C22:6 和 100μM C18:2 补充导致第 7 天胚胎存活率降低（P<0.05）。C18:3 补充对第 7 天胚胎存活率没有影响，但与 100μM β-ME 补充相比，100μM C18:3 增加了第 8 天胚胎存活率（P<0.05）。实验 3 和 4：与所有其他补充相比，10μM C22:6 补充后硬脂酰辅酶 A 去饱和酶 1（SCD1）和固醇调节元件结合转录因子 1（SREBP1）mRNA 减少（P<0.05）。与没有脂肪酸补充的组相比，发现 100μM C18:2、10μM C22:6 和 10μM C18:3 补充后脂肪酸去饱和酶 2（FADS2）转录水平降低（P<0.05）。乙酰辅酶 A 羧化酶（ACC）、脂肪酸合酶（FAS）、脂肪分化相关蛋白（ADRP）、长链酰基辅酶 A 合成酶家族成员 1（ACSL1）、二酰基甘油 O-酰基转移酶 1（DGAT1）、肉碱棕榈酰基转移酶-II（CPT-II）mRNA 表达和 AMPKalpha 磷酸化不受 PUFA 补充的影响。实验 5：与血清补充相比，Boec 组 SCD1 和 FAS mRNA 减少，与母羊输卵管组 SCD1 mRNA 减少（P<0.05）。综上所述，本研究表明，在牛培养 6 天过程中补充 C18:2、C18:3 或 C22:6 以及与 Boec 共培养可下调第 7-8 天胚胎中参与脂质代谢的蛋白质的 mRNA 表达（SCD1 和 FADS2 去饱和酶），可能是通过 10μM C22:6 补充后的 SREBP1 mRNA 调节，表明牛囊胚中饱和/不饱和脂肪酸平衡发生了变化。
