甘氨酸对猪卵母细胞体外成熟质量及卵丘细胞的影响

本实验旨在研究甘氨酸对卵丘细胞及卵母细胞体外成熟质量的影响,优化猪卵母细胞体外成熟(IVM)培养体系。在猪卵母细胞体外成熟培养液中添加6 mmol/L甘氨酸,体外培养至44 h后,通过倒置显微镜测量卵丘细胞扩散直径,结合流式细胞术与qPCR检测卵丘细胞凋亡情况;进一步检测成熟卵母细胞中谷胱甘肽(GSH)、细胞成熟促进因子(MPF)和丝裂原活化蛋白激酶(MAPK)水平及细胞成熟相关基因表达。结果表明:甘氨酸能显著提高卵丘细胞扩散直径,减少卵丘细胞的总凋亡率,并能显著提高卵丘细胞中Bcl-2的mRNA表达;甘氨酸能显著增加成熟卵母细胞中的GSH、MPF和MAPK水平以及成熟相关基因BMP15、GDF9、CyclinB1、CDK1、C-MOS的mRNA表达。综上可知,甘氨酸可提高猪卵母细胞体外成熟质量,降低卵丘细胞凋亡率。

Effect of Glycine on in vitro Maturation Quality of Pig Oocytes and Cumulus Cells

In order to optimize the culture system of porcine oocytes in vitro maturation(IVM), the effects of glycine on cumulus cells and the quality of in vitro maturation oocytes were investigated. A total of 6 mmol/L glycine was added into the IVM medium of porcine oocytes. The cumulus cells diffusing diameter were measured before and after oocytes culture, and the cumulus cells diffusing were compared. The apoptosis of cumulus cells were detected by flow cytometry after Annexin V-FITC staining, and the cumulus cells apoptosis related genes were detected by qPCR. In order to investigate the effect of glycine on the quality of oocytes maturation, the levels of GSH, MPF, MAPK and the expression of genes related to oocytes maturation were detected by qPCR. The results show that glycine group could significantly increase the diffusing diameter of cumulus cells after IVM, reduce the total apoptotic rate of cumulus cells, and significantly increase the expression of Bcl2 in cumulus cells(P<0.05). Glycine could significantly increase the GSH, MPF and MAPK in mature oocyte(P<0.05), and the expression of mature related genes BMP15, GDF9, CyclinB1, CDK1 and C-MOS were significantly up-regulated(P<0.05). In conclusion, glycine can improve the quality of in vitro maturation porcine oocytes and reduce the apoptosis rate of cumulus cells.