CAJ | 학술논문

目的：通过检测玻璃化冷冻前后卵母细胞线粒体DNA（mtDNA）缺失和能量代谢变化，以探讨冷冻保存对卵母细胞质量的影响。方法：收集体外受精（IVF）废弃的人卵母细胞和受精卵，包括未受精（0PN）、两原核（2PN）和三原核（3PN）；随机分为2组，A组为经玻璃化冷冻保存，B组为新鲜对照组。采用巢式聚合酶链反应（nest PCR）检测冷冻前后卵母细胞mtDNA缺失差异；并进一步比较冷冻前后卵母细胞mtDNA拷贝数和三磷酸腺苷（ATP）含量变化。结果：卵母细胞mtDNA缺失率和mtDNA拷贝数在冷冻处理前后差异无统计学意义（P＞0.05）；冷冻后卵母细胞内ATP含量低于冷冻前新鲜卵母细胞。结论：冷冻保存对卵母细胞mtDNA缺失及拷贝数影响较小，可较好地实现保存卵母细胞的目的。
목적：통과검측파리화냉동전후란모세포선립체DNA（mtDNA）결실화능량대사변화，이탐토냉동보존대란모세포질량적영향。방법：수집체외수정（IVF）폐기적인란모세포화수정란，포괄미수정（0PN）、량원핵（2PN）화삼원핵（3PN）；수궤분위2조，A조위경파리화냉동보존，B조위신선대조조。채용소식취합매련반응（nest PCR）검측냉동전후란모세포mtDNA결실차이；병진일보비교냉동전후란모세포mtDNA고패수화삼린산선감（ATP）함량변화。결과：란모세포mtDNA결실솔화mtDNA고패수재냉동처리전후차이무통계학의의（P＞0.05）；냉동후란모세포내ATP함량저우냉동전신선란모세포。결론：냉동보존대란모세포mtDNA결실급고패수영향교소，가교호지실현보존란모세포적목적。
Objective: To explore the effect of cryopreservation on the mitochondrial DNA (mtDNA) deletions and energy metabolism of human oocyte, so as to investigate the effect of cryopreservation on oocytes preservation. Methods: The dumped oocytes with 0PN, 2PN and 3PN during routine IVF were collected for this cryopreservation study. Those oocytes were randomly divided into two groups, one for vitrification cryopreservation (Group A) and the other as control (Group B, without cryopreservation). The nested polymerase chain reaction (PCR) was used to detect the mtDNA deletions of oocyte mtDNA before and after cryopreservation. Real-time quantitative PCR was adopted to determine the copy number of oocyte mtDNA. ATP contents were detected using luciferin-luciferase assays. Results: There were not significant differences in the mutation frequency of oocyte mtDNA deletions and the copy number of mtDNA between two groups (P>0.05). However, The ATP content in the group A was significantly lower than that in the group B. Conclusions: The effect of cryopreservation on the oocyte mtDNA deletions and the copy number of mtDNA was limited, suggesting that cryopreservation can be properly used to preserve oocyte.