Elahi F, Lee H, Lee J, Lee S T, Park C K, Hyun S-H, Lee E
Laboratory of Theriogenology, College of Veterinary Medicine, Kangwon National University, Chuncheon, Korea.
Institute of Veterinary Science, Kangwon National University, Chuncheon, Korea.
Reprod Domest Anim. 2017 Oct;52(5):741-748. doi: 10.1111/rda.12974. Epub 2017 Apr 10.
This study investigated the effects of early induction of autophagy on embryonic development in pigs. For this, oocytes or embryos were treated with an autophagy inducer, rapamycin (RP), during post-activation (Pa), in vitro fertilization (IVF) and/or in vitro culture (IVC). When parthenogenesis (PA) embryos were untreated (control) or treated with various concentrations of RP for 4 hr during Pa, 100 nm RP showed a higher blastocyst formation (48.8 ± 2.7%) than the control (34.6 ± 3.0%). When PA embryos were treated during the first 24 hr of IVC, blastocyst formation was increased (p < .05) by 1 and 10 nm RP (61.9 ± 3.0 and 59.6 ± 3.0%, respectively) compared to the control (43.2 ± 1.8%) and 100 nm RP (47.8 ± 3.2%), with a higher embryo cleavage in response to 10 nm RP (87.3 ± 2.4%) than the control (74.1 ± 3.2%). RP treatment during IVC and Pa + IVC showed increased blastocyst formation (44.7 ± 2.5 and 44.1 ± 2.0%, respectively) compared to the control (33.2 ± 2.0%). In addition, RP treatment during Pa and/or IVC increased glutathione content and inversely reduced reactive oxygen species. In IVF, RP treatment for 6 hr during IVF significantly increased embryonic development (34.0 ± 2.6%) compared to the control (24.8 ± 1.6%), but treatment during IVC for 24 hr with RP did not (23.0 ± 3.8%). Autophagy was significantly increased in PA oocytes by the RP treatment during Pa but not altered by the treatment during the first 24 hr of IVC. Overall, RP treatment positively regulated the pre-implantation development of pig embryos, probably by regulating cellular redox state and stimulating autophagy.
本研究调查了早期诱导自噬对猪胚胎发育的影响。为此,在激活后(Pa)、体外受精(IVF)和/或体外培养(IVC)期间,用自噬诱导剂雷帕霉素(RP)处理卵母细胞或胚胎。当孤雌生殖(PA)胚胎在Pa期间未处理(对照)或用不同浓度的RP处理4小时时,100纳米的RP显示出比对照(34.6±3.0%)更高的囊胚形成率(48.8±2.7%)。当PA胚胎在IVC的前24小时接受处理时,与对照(43.2±1.8%)和100纳米的RP(47.8±3.2%)相比,1纳米和10纳米的RP使囊胚形成率增加(p<0.05)(分别为61.9±3.0%和59.6±3.0%),且10纳米的RP诱导的胚胎分裂率(87.3±2.4%)高于对照(74.1±3.2%)。在IVC以及Pa+IVC期间进行RP处理,与对照(33.2±2.0%)相比,囊胚形成率增加(分别为44.7±2.5%和44.1±2.0%)。此外,在Pa和/或IVC期间进行RP处理可增加谷胱甘肽含量,并反向降低活性氧水平。在IVF中,与对照(24.8±1.6%)相比,在IVF期间用RP处理6小时可显著提高胚胎发育率(34.0±2.6%),但在IVC期间用RP处理24小时则不然(23.0±3.8%)。在Pa期间用RP处理可使PA卵母细胞中的自噬显著增加,但在IVC的前24小时进行处理则不会改变自噬水平。总体而言,RP处理可能通过调节细胞氧化还原状态和刺激自噬,对猪胚胎植入前发育起到正向调节作用。