Mesalam Ayman, Khan Imran, Lee Kyeong-Lim, Song Seok-Hwan, Chowdhury M M R, Uddin Zia, Park Ki Hun, Kong Il-Keun
Department of Animal Science, Division of Applied Life Science (BK21 Plus), Gyeongsang National University, Jinju, 660-701, Gyeongnam Province, Republic of Korea; Department of Theriogenology, Faculty of Veterinary Medicine, Zagazig University, 44519, Egypt.
Department of Animal Science, Division of Applied Life Science (BK21 Plus), Gyeongsang National University, Jinju, 660-701, Gyeongnam Province, Republic of Korea.
Theriogenology. 2017 Sep 1;99:10-20. doi: 10.1016/j.theriogenology.2017.05.012. Epub 2017 May 19.
2-Methoxystypandrone (2-MS), a naphthoquinone from Polygonum cuspidatum, has been reported to exhibit antioxidant activities. However, its pharmacological properties have not been fully studied in vitro culture system of oocytes and embryos development. In this study, we examined the effects of 2-MS on bovine blastocyst developmental competency during preimplantation using TUNEL assay, qRT-PCR and immunocytochemistry. Oocytes were treated with 1 μM 2-MS during maturation, followed by in vitro fertilized and the zygotes were cultured. We observed that blastocyst development was significantly higher in the 2-MS treated oocytes (45.29%) compared to control groups (32.21%). We also found that 2-MS treatment significantly increased total cell number and decreased the apoptotic cells in Day-8 blastocyst. Finally, we examined the signal pathway affecting the embryo development after the 2-MS treatment. We detected a significant decrease in the mRNA levels of NFKB1, IKBKB, COX2, NOS2, BAX, CASP3, and JAK2 after 2-MS treatment, however, the mRNA level of the anti-apoptotic gene BCL2 was significantly higher than that in the control. Moreover, protein expression levels of NFKB1, IKBKB, and COX2, and 8-oxoG DNA lesion decreased significantly in the 2-MS treatment group, compared to the control. In summary, our findings suggested that treatment of oocytes with a potential anti-oxidant, 2-MS during IVM improved in vitro developmental competence of bovine embryos.
2-甲氧基斯台潘酮（2-MS）是一种来自虎杖的萘醌，据报道具有抗氧化活性。然而，其药理特性在卵母细胞和胚胎发育的体外培养系统中尚未得到充分研究。在本研究中，我们使用TUNEL检测、qRT-PCR和免疫细胞化学方法，研究了2-MS对牛植入前胚泡发育能力的影响。卵母细胞在成熟过程中用1μM 2-MS处理，随后进行体外受精并培养受精卵。我们观察到，与对照组（32.21%）相比，经2-MS处理的卵母细胞的胚泡发育率显著更高（45.29%）。我们还发现，2-MS处理显著增加了第8天胚泡的总细胞数并减少了凋亡细胞。最后，我们研究了2-MS处理后影响胚胎发育的信号通路。2-MS处理后，我们检测到NFKB1、IKBKB、COX2、NOS2、BAX、CASP3和JAK2的mRNA水平显著降低，然而，抗凋亡基因BCL2的mRNA水平显著高于对照组。此外，与对照组相比，2-MS处理组中NFKB1、IKBKB和COX2的蛋白表达水平以及8-氧代鸟嘌呤DNA损伤显著降低。总之，我们的研究结果表明，在体外成熟过程中用潜在的抗氧化剂2-MS处理卵母细胞可提高牛胚胎的体外发育能力。
