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目的探讨体外受精-胚胎移植周期卵丘细胞乙酰辅酶A酰基转移酶2(acetyl-coenzyme A acyltransferase 2protein,ACAA2)表达与卵丘细胞内活性氧水平的关系及其对胚胎发育的影响。方法 49例体外授精-胚胎移植患者,收集其卵丘细胞,采用荧光探针二氯荧光黄双乙酸盐检测卵丘细胞内活性氧水平,采用实时荧光PCR法检测卵丘细胞内ACAA2mRNA水平。取卵后第3天选择良好胚胎进行移植,胚胎移植后14d测定血β-人绒毛膜促性腺激素(human chorionic gonadotrophin,HCG)水平,血β-HCG≥50u/L者为妊娠组,血β-HCG<50u/L者为未妊娠组,比较2组ACAA2基因相对表达量,分析卵丘细胞ACAA2 mRNA表达水平与细胞内活性氧水平及优质胚胎形成率的关系。结果卵丘细胞活性氧水平为187.091 2±71.131 1,ACAA2mRNA水平为1.211 9±0.987 3;49例优质胚胎形成率为(31.458 2±5.871 2)%;妊娠组27例,未妊娠组22例,妊娠组ACAA2基因相对表达量(1.548 6±0.993 9)与未妊娠组(0.849 2±0.339 5)比较差异无统计学意义(P>0.05);卵丘细胞ACAA2mRNA表达水平与卵丘细胞内活性氧水平呈负相关(r=-0.712,P=0.009),与优质胚胎形成率呈正相关(r=0.471,P=0.036)。结论卵丘细胞内ACAA2可能参与卵泡内脂肪代谢,并抑制细胞内活性氧水平过高,进而控制卵泡液活性氧水平,保护卵母细胞免受氧化应激损伤。
Objective To investigate the relationship between the expression of acetyl-coenzyme A acyltransferase 2 protein (ACAA2) and the level of reactive oxygen species (ROS) in cumulus cells during the in vitro fertilization-embryo transfer and its effect on embryonic development. Methods 49 cases of in vitro fertilization and embryo transfer were collected from patients with cumulus cells. The level of reactive oxygen species (ROS) in cumulus cells was detected by fluorescent probe dichloro-fluorescein diacetate. The level of ACAA2 mRNA in cumulus cells was detected by real-time fluorescence PCR. On the 3rd day after ovulation, good embryos were selected for transplantation. The level of serum human chorionic gonadotrophin (HCG) was measured 14 days after embryo transfer, and the levels of serum β-HCG≥50 u / L in pregnancy group and blood β -HCG <50u / L for the non-pregnant group, the relative expression of ACAA2 gene in two groups were compared, and the relationship between ACAA2 mRNA expression level and intracellular reactive oxygen species and high-quality embryogenesis rate was analyzed. Results The levels of reactive oxygen species in cumulus cells were 187.091 2 ± 71.131 1 and ACAA2 mRNA levels were 1.211 9 ± 0.987 3. The rate of high-quality embryo formation in 49 cases was (31.458 2 ± 5.871 2)%. In pregnancy group (n = 27) and non-pregnancy group (n = 22) The relative expression level of ACAA2 gene in pregnancy group (1.548 6 ± 0.993 9) and non-pregnancy group (0.849 2 ± 0.339 5) showed no significant difference (P> 0.05). The expression level of ACAA2 mRNA in cumulus cells was similar to that of reactive oxygen species (R = -0.712, P = 0.009), which was positively correlated with the rate of high quality embryo formation (r = 0.471, P = 0.036). Conclusion ACAA2 in the cumulus cells may be involved in the metabolism of fat in the follicles and inhibit the excessive level of reactive oxygen species in the follicles, thereby controlling the level of reactive oxygen species in follicular fluid and protecting the oocytes from oxidative stress.