Sun Ling, Lin Tao, Lee Jae Eun, Kim So Yeon, Bai Ying, Jin Dong Il
School of Life Sciences and Food Engineering, Hebei University of Engineering, Handan 056038, China.
Division of Animal & Dairy Science, Chungnam National University, Daejeon 34134, Korea.
J Anim Sci Technol. 2024 Jul;66(4):726-739. doi: 10.5187/jast.2024.e68. Epub 2024 Jul 31.
This study was conducted to investigate whether lysophosphatidic acid (LPA) could improve the development of porcine somatic cell nuclear transfer (SCNT) embryos. Porcine SCNT-derived embryos were cultured in chemically defined polyvinyl alcohol (PVA)-based porcine zygote medium (PZM)-4 without or with LPA, and the development, cell proliferation potential, apoptosis, and expression levels of pluripotent markers were evaluated. LPA significantly increased the rates of cleavage and blastocyst formation compared to those seen in the LPA un-treatment (control) group. The expression levels of embryonic development-related genes (IGF2R, PCNA and CDH1) were higher ( < 0.05) in the LPA treatment group than in the control group. LPA significantly increased the numbers of total, inner cell mass and EdU (5-ethynyl-2'-deoxyuridine)-positive cells in porcine SCNT blastocysts compared to those seen in the control group. TUNEL assay showed that LPA significantly reduced the apoptosis rate in porcine SCNT-derived embryos; this was confirmed by decreases ( < 0.05) in the expression levels of pro-apoptotic genes, BAX and CASP3, and an increase (p < 0.05) in the expression level of the anti-apoptotic gene, BCL2L1. In addition, LPA significantly increased Oct4 expression at the gene and protein levels. Together, our data suggest that LPA improves the quality and development of porcine SCNT-derived embryos by reducing apoptosis and enhancing cell proliferation and pluripotency.
本研究旨在探讨溶血磷脂酸(LPA)是否能促进猪体细胞核移植(SCNT)胚胎的发育。将猪SCNT胚胎在不含或含有LPA的化学限定的基于聚乙烯醇(PVA)的猪合子培养基(PZM)-4中培养,并评估其发育、细胞增殖潜能、凋亡情况以及多能性标志物的表达水平。与未处理LPA(对照组)的情况相比,LPA显著提高了卵裂率和囊胚形成率。LPA处理组中胚胎发育相关基因(IGF2R、PCNA和CDH1)的表达水平高于对照组(P<0.05)。与对照组相比,LPA显著增加了猪SCNT囊胚中的总细胞数、内细胞团细胞数以及5-乙炔基-2'-脱氧尿苷(EdU)阳性细胞数。TUNEL检测显示,LPA显著降低了猪SCNT胚胎的凋亡率;促凋亡基因BAX和CASP3表达水平的降低(P<0.05)以及抗凋亡基因BCL2L1表达水平的升高(P<0.05)证实了这一点。此外,LPA在基因和蛋白水平上均显著增加了Oct4的表达。总之,我们的数据表明,LPA通过减少凋亡、增强细胞增殖和多能性来提高猪SCNT胚胎的质量和发育能力。