摘要
目的研究多囊卵巢综合征(PCOS)不孕患者全血N6-甲基腺嘌呤(m6A)及相关蛋白mRNA表达与体外受精-胚胎移植(IVF-ET)妊娠结局的关系。方法选取2021年1—12月在宁波大学附属妇女儿童医院生殖中心接受IVF-ET的80例PCOS不孕患者作为辅助生育组,选取同期体检的80例健康女性作为对照组。采用比色法检测两组全血m6A水平,采用qRT-PCR检测全血FTO、ALKBH5、YTHDF2 mRNA相对水平,检测两组血清睾酮(T)、黄体生成素(LH)、卵泡刺激素(FSH)、雌二醇(E_(2))水平。根据移植后6~8周超声结果分为妊娠成功组与妊娠失败组,收集两组临床资料,采用多因素logistic回归模型分析妊娠失败的危险因素。采用受试者工作特征(ROC)曲线分析m6A及相关蛋白mRNA表达对妊娠失败的预测价值。结果辅助生育组全血m6A表达量[(0.07±0.02)ng]低于对照组[(0.11±0.02)ng],差异有统计学意义(t=11.157,P<0.05),FTO mRNA[1.26(0.82,2.27)]、ALKBH5 mRNA[1.08(0.53,1.85)]相对表达量均高于对照组[0.85(0.45,1.52),0.81(0.41,1.42)],差异均有统计学意义(Z=-3.744,-2.061,均P<0.05)。80例患者经首次IVF-ET辅助生育治疗后,妊娠成功43例,妊娠失败37例。妊娠成功组与妊娠失败组促性腺激素(Gn)用量[(1852.55±454.85)IU vs.(1946.28±403.85)IU,t=2.071,P<0.05]、获卵总数[(12.40±5.01)个vs.(10.84±6.80)个,t=2.502,P<0.05]、种植率[80.30%(53/66)vs.11.11%(6/54),χ^(2)=56.890,P<0.05]、m6A表达量[(0.08±0.02)ng vs.(0.06±0.01)ng,t=-5.329,P<0.05]、FTO mRNA[1.08(0.70,1.70)vs.1.77(0.90,3.04),Z=-2.716,P<0.05]、ALKBH5 mRNA[0.77(0.53,1.51)vs.1.59(0.57,2.98),Z=-2.244,P<0.05]相对表达量比较,差异均有统计学意义。多因素logistic回归分析结果显示:m6A(OR=1.135,95%CI:1.045~1.307,P<0.05)、FTO mRNA(OR=1.738,95%CI:1.225~2.274,P<0.05)、ALKBH5 mRNA(OR=1.979,95%CI:1.383~2.701,P<0.05)相对表达量为PCOS不孕患者妊娠失败的危险因素。ROC曲线显示:m6A、FTO mRNA、ALKBH5mRNA联合预测的曲线下面积(AUC)为0.806(95%CI:0.710~0.903),灵敏度与特异度分别为93.0%、64.9%。结论PCOS不孕患者全血样本中m6A表达下调,FTO mRNA、ALKBH5 mRNA表达上调,是患者妊娠结局的影响因素,可望作为早期评估妊娠结局的有效标志物。
Objective To study the correlations between whole blood N6-methyladenine(m6A)and related protein mRNA expression with pregnancy outcome after in vitro fertilization-embryo transfer(IVF-ET)in infertile patients with polycystic ovary syndrome(PCOS).Methods Eighty infertile PCOS patients who underwent IVF-ET in Reproductive Center of Women and Children's Hospital of Ningbo University were selected as assisted reproductive group,and 80 healthy women who underwent physical examination in the hospital during the same period were selected as control group.The levels of m6A in whole blood of the two groups were detected by colorimetry,and the relative levels of FTO mRNA,ALKBH5 mRNA,YTHDF2 mRNA in whole blood samples were detected by qRT-PCR.The serum levels of testosterone(T),luteinizing hormone(LH),follicle stimulating hormone(FSH),and estradiol(E_2)in the two groups were detected.According to ultrasound results at 6-8 weeks after transplantation,the patients were divided into pregnancy success group and pregnancy failure group.The clinical data of the two groups were collected.The risk factors of pregnancy failure were analyzed by multivariate logistic regression model.The predictive value of m6A and related protein mRNA expression for pregnancy failure was analyzed by receiver operating characteristic(ROC)curve.Results The expression level of m6A in whole blood of assisted reproductive group[(0.07±0.02)ng]was lower than that of normal control group[(0.11±0.02)ng],there was statistically significant difference(t=11.157,P<0.05),the relative expression levels of FTO mRNA[1.26(0.82,2.27)]and ALKBH5 mRNA[1.08(0.53,1.85)]were higher than those of normal control group[0.85(0.45,1.52),0.81(0.41,1.42)],there were statistically significant differences(Z=-3.744,-2.061,P<0.05).After the first IVF-ET assisted fertility treatment in 80 patients,43 patients succeeded,and 37 patients failed.There were statistically significant differences in gonadotropin(Gn)dose[(1852.55±454.85)IU vs.(1946.28±403.85)IU,t=2.071,P<0.05],total number of retrieved eggs[(12.40±5.01)vs.(10.84±6.80),t=2.502,P<0.05],implantation rate[80.30%(53/66)vs.11.11%(6/54),χ~2=56.890,P<0.05],expression of m6A[(0.08±0.02)ng vs.(0.06±0.01)ng,t=-5.329,P<0.05],relative expression of FTO mRNA[1.08(0.70,1.70)vs.1.77(0.90,3.04),Z=-2.716,P<0.05],and relative expression of ALKBH5 mRNA[0.77(0.53,1.51)vs.1.59(0.57,2.98),Z=-2.244,P<0.05]between pregnancy success group and pregnancy failure group.Multivariate logistic regression analysis showed that expression of m6A(OR=1.135,95%CI:1.045-1.307,P<0.05),relative expression of FTO mRNA(OR=1.738,95%CI:1.225-2.274,P<0.05),and relative expression of ALKBH5 mRNA(OR=1.979,95%CI:1.383-2.701,P<0.05)were the risk factors of pregnancy failure in PCOS infertile patients.ROC curve showed that the area under the curve(AUC)predicted by m6A,FTO mRNA,and ALKBH5 mRNA was 0.806(95%CI:0.710-0.903),the sensitivity and specificity were 93.0%and 64.9%,respectively.Conclusion The down-regulation of m6A and up-regulation of FTO mRNA and ALKBH5 mRNA expressions are influencing factors of pregnancy outcome in patients with PCOS-related infertility,so above indicators can be used as effective markers for early assessment of pregnancy outcome.
作者
李卓玲
卢琳
蔡婕
郑乐朋
周黎明
LI Zhuo-ling;LU Lin;CAI Jie;ZHENG Le-peng;ZHOU Li-ming(Department of Laboratory,Women and Children's Hospital of Ningbo University,Ningbo,Zhejiang 315012,China)
出处
《中国妇幼保健》
CAS
2024年第24期4898-4901,共4页
Maternal and Child Health Care of China
基金
浙江省医药卫生科技计划项目(2021KY1052)。
作者简介
通信作者:周黎明,E-mail:zhou.li.ming@163.com。
