印记基因SNRPN与不明原因复发性流产的相关性研究

Correlation Study of Imprinted Gene SNRPN and Unexplained Recurrent Miscarriage

摘要: 目的：探讨印记基因SNRPN的表达、单核苷酸多态性(single nucleotide polymorphism, SNP)及其启动子区甲基化状态与不明原因复发性流产(unexplained recurrent spontaneous abortion ,URSA)的相关性。方法：选择2021年7月至2023年2月本院31例不明原因复发性流产患者绒毛组织(URSA组)和34例正常妊娠自愿流产患者的绒毛组织(对照组)为研究对象。采用实时荧光定量PCR (qRT-PCR)检测两组绒毛组织中SNRPN的mRNA表达水平；通过一代(Sanger)测序分析SNRPN多个SNP位点的差异；利用甲基化转化试剂盒结合PyroMark Q96实时定量焦磷酸系列分析仪检测SNRPN基因启动子区的甲基化状态。结果：1) URSA组绒毛组织中SNRPN的相对表达量显著高于对照组(0.55 ± 0.81 vs. 0.17 ± 0.16，P < 0.05)。2) URSA组和对照组的SNRPN基因rs220030位点基因型、等位基因分布差异有统计学意义(P < 0.05)，而rs7164989、rs12905653、rs705位点差异无统计学意义(P > 0.05)。3) 甲基化检测结果显示，URSA组SNRPN基因启动子区的甲基化水平显著低于对照组(P < 0.05)，低甲基化状态可能通过增加SNRPN基因的表达水平，进而促进URSA的发生。结论：URSA患者绒毛组织中SNRPN基因的高表达可能与其启动子区低甲基化状态有关，而rs220030位点的单核苷酸多态性可能与URSA的发生密切相关。这些发现为URSA的发病机制提供了新的视角，并为临床诊断和治疗提供了潜在的生物标志物。

Abstract: Purpose: To explore the expression of the imprinted gene SNRPN and single nucleotide polymorphism the correlation between SNP and the methylation status of its promoter region and unexplained recurrent spontaneous abortion (URSA). Methods: The chorionic villus tissues of 31 patients with unexplained recurrent miscarriage (URSA group) and 34 patients with normal pregnancy and voluntary miscarriage (control group) in our hospital from July 2021 to February 2023 were selected as the research subjects. The mRNA expression levels of SNRPN in the villus tissues of the two groups were detected by real-time fluorescence quantitative PCR (qRT-PCR). The differences of multiple SNP loci of SNRPN were analyzed through first-generation (Sanger) sequencing; The methylation status of the promoter region of the SNRPN gene was detected by using the methylation conversion kit combined with the PyroMark Q96 real-time quantitative pyrophosphate series analyzer. Results: 1) The relative expression level of SNRPN in the villous tissue of the URSA group was significantly higher than that of the control group (0.55 ± 0.81 vs. 0.17 ± 0.16, P < 0.05). 2) There was a statistically significant difference in the genotype and allele distribution of the SNRPN gene at the rs220030 locus between the URSA group and the control group (P < 0.05), while there was no statistically significant difference at the rs7164989, rs12905653, and rs705 loci (P > 0.05). 3) The methylation detection results showed that the methylation level in the promoter region of the SNRPN gene in the URSA group was significantly lower than that in the control group (P < 0.05). The hypomethylation state might promote the occurrence of URSA by increasing the expression level of the SNRPN gene. Conclusion: The high expression of the SNRPN gene in the villous tissues of patients with URSA may be related to the hypomethylation status of its promoter region, while the single nucleotide polymorphism at the rs220030 locus may be closely related to the occurrence of URSA. These findings provide a new perspective on the pathogenesis of URSA and offer potential biomarkers for clinical diagnosis and treatment.