Hwang Seon-Ung, Kim Kyu-Jun, Kim Eunhye, Yoon Junchul David, Park Kyu Mi, Jin Minghui, Han Yongquan, Kim Mirae, Lee Gabsang, Hyun Sang-Hwan
Laboratory of Veterinary Embryology and Biotechnology, Veterinary Medical Center and College of Veterinary Medicine, Chungbuk National University, Cheongju, Republic of Korea; Institute of Stem Cell & Regenerative Medicine, Chungbuk National University, Cheongju, Republic of Korea.
Institute for Cell Engineering, Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
Theriogenology. 2018 Jun;113:197-207. doi: 10.1016/j.theriogenology.2018.02.020. Epub 2018 Mar 2.
Lysophosphatidic acid (LPA) is a phospholipid-derived signaling molecule with biological activities, such as stimulating cell proliferation, differentiation and migration. In the present study, we examined the effect of LPA on porcine oocytes during in vitro maturation (IVM) and subsequent embryonic development following parthenogenetic activation (PA) and in vitro fertilization (IVF). During IVM, the maturation medium was supplemented with various concentrations of LPA (0, 10, 30, and 60 μM). After 42 h of IVM, the 30 μM LPA-treated group showed a significant (P <0.05) increase in nuclear maturation and intracellular glutathione (GSH) levels compared with the other groups. The 30 μM LPA-treated group exhibited a significant decrease in intracellular reactive oxygen species (ROS) levels compared with the other groups. In PA, the 30 μM LPA-treated group had significantly higher cleavage (CL) and blastocyst (BL) rates compared with those of the other LPA-treated groups. In IVF, the 30 μM LPA-treated group had significantly higher CL and BL rates than the other LPA-treated groups. The expression of the developmental competence gene (proliferating cell nuclear antigen, PCNA) in the oocytes and cumulus cells of the individuals in the 30 μM LPA-treated group was significantly increased compared with the control group. In addition, the specific expression of urokinase Plasminogen Activator (uPA) and uPA Receptor (uPAR) in cumulus cells was significantly increased in the 30 μM LPA-treated group. The western blotting results revealed that LPA improves the activities of p38 mitogen-activated protein kinase (MAPK) and epidermal growth factor (EGF) by enhanced phosphorylation. In conclusion, treatment with 30 μM LPA during IVM promotes enhances the EGF-EGFR signaling pathway, resulting in cumulus cell expansion. And then, this treatment improves the developmental potential of PA and IVF porcine embryos by enhancing nuclear and cytoplasmic maturation and reducing ROS.
溶血磷脂酸（LPA）是一种具有生物活性的磷脂衍生信号分子，具有刺激细胞增殖、分化和迁移等生物活性。在本研究中，我们检测了LPA对猪卵母细胞体外成熟（IVM）以及孤雌激活（PA）和体外受精（IVF）后后续胚胎发育的影响。在IVM期间，成熟培养基中添加了不同浓度的LPA（0、10、30和60μM）。IVM 42小时后，与其他组相比，30μM LPA处理组的核成熟和细胞内谷胱甘肽（GSH）水平显著（P<0.05）升高。与其他组相比，30μM LPA处理组的细胞内活性氧（ROS）水平显著降低。在PA中，与其他LPA处理组相比，30μM LPA处理组的卵裂（CL）率和囊胚（BL）率显著更高。在IVF中，30μM LPA处理组的CL和BL率显著高于其他LPA处理组。与对照组相比，30μM LPA处理组个体的卵母细胞和卵丘细胞中发育能力基因（增殖细胞核抗原，PCNA）的表达显著增加。此外，30μM LPA处理组卵丘细胞中尿激酶型纤溶酶原激活剂（uPA）和uPA受体（uPAR）的特异性表达显著增加。蛋白质印迹结果显示，LPA通过增强磷酸化提高了p38丝裂原活化蛋白激酶（MAPK）和表皮生长因子（EGF）的活性。总之，IVM期间用30μM LPA处理可促进增强EGF-EGFR信号通路，导致卵丘细胞扩展。然后，这种处理通过增强核成熟和细胞质成熟以及降低ROS来提高PA和IVF猪胚胎的发育潜力。
