Family Planning Research Institute and Center for Reproductive Medicine, Tongji Medical College, Huazhong University of Science and Technology, Wuhan,, 430030, People's Republic of China.
Reproductive Medicine Center, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, People's Republic of China.
Reprod Biol Endocrinol. 2019 Jul 10;17(1):54. doi: 10.1186/s12958-019-0495-6.
Cell-free mitochondrial DNA (cf-mtDNA) in body fluids has attracted much attention for the purpose of monitoring disease because of the clinical advantages. This study investigated whether the cf-mtDNA content in human follicular fluid samples was associated with oocyte and embryo developmental competence.
We collected 225 individual follicular fluid samples from 92 patients undergoing conventional in vitro fertilization (n = 53) or intracytoplasmic sperm injection (n = 39). cf-mtDNA and cell-free nuclear DNA (cf-nDNA) were measured using real-time quantitative PCR for the ND1 and β-globin genes. Multivariate logistic regression and linear regression were used to analyze data.
The relative cf-mtDNA content (cf-ND1/cf-β-globin ratio) in follicular fluid was significantly lower in the group showing blastocyst development than in the non-blastocyst group (P = 0.030). Additionally, the relative cf-mtDNA content was significantly and positively correlated with the age of the female patient (P = 0.009), while the relative cf-mtDNA content for older women (≥38 years old) with anti-Müllerian hormone (AMH) ≤1.1 ng/ml was significantly higher than in those with AMH > 1.1 ng/ml (P <0.05). The cf-nDNA content was significantly positively correlated with the antral follicle count (P = 0.012), and significantly negatively correlated with both the number of days of stimulation and the total dose of gonadotropin administration (P = 0.039 and P = 0.015, respectively). Neither cf-mtDNA nor cf-nDNA levels in follicular fluid were associated with oocyte maturation, fertilization, or Day 3 embryo morphological scoring.
The relative cf-mtDNA content in human follicular fluid was negatively correlated with blastulation and positively correlated with the patient age, indicating that it is a promising bio-marker to evaluate oocyte developmental competence.
由于具有临床优势，体液中的无细胞线粒体 DNA（cf-mtDNA）已引起人们对监测疾病的关注。本研究旨在探讨人卵泡液样本中的 cf-mtDNA 含量是否与卵母细胞和胚胎发育能力有关。
我们收集了 92 名接受常规体外受精（n=53）或胞浆内精子注射（n=39）的患者的 225 个单个卵泡液样本。使用实时定量 PCR 测量 ND1 和 β-球蛋白基因的 cf-mtDNA 和无细胞核 DNA（cf-nDNA）。采用多元逻辑回归和线性回归分析数据。
与未形成囊胚的组相比，形成囊胚的组卵泡液中的相对 cf-mtDNA 含量（cf-ND1/cf-β-球蛋白比值）显著降低（P=0.030）。此外，相对 cf-mtDNA 含量与女性患者的年龄呈显著正相关（P=0.009），而抗苗勒管激素（AMH）≤1.1ng/ml 的年龄较大的女性（≥38 岁）的相对 cf-mtDNA 含量明显高于 AMH＞1.1ng/ml 的女性（P＜0.05）。cf-nDNA 含量与窦卵泡计数呈显著正相关（P=0.012），与促性腺激素给药天数和总剂量呈显著负相关（P=0.039 和 P=0.015）。卵泡液中的 cf-mtDNA 或 cf-nDNA 水平均与卵母细胞成熟、受精或第 3 天胚胎形态评分无关。
人卵泡液中的相对 cf-mtDNA 含量与囊胚形成呈负相关，与患者年龄呈正相关，表明其是评估卵母细胞发育能力的有前途的生物标志物。
