Department of Obstetrics and Gynecology, Center for Reproductive Medicine, Qilu Hospital, Cheeloo College of Medicine, Shandong University, Jinan, Shandong 250012, P.R. China.
Department of Obstetrics and Gynecology, Third Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325200, P.R. China.
Mol Med Rep. 2022 Feb;25(2). doi: 10.3892/mmr.2021.12559. Epub 2021 Dec 8.
Recurrent pregnancy loss (RPL) is often associated with dysregulated Annexin A5 (ANXA5) expression. Moreover, the variants of Anxa5, a protein that is enriched in the placenta to prevent coagulation, have been reported to affect RPL risks. The haplotypes M1 [including single nucleotide polymorphisms (SNPs) 1A/C and 27T/C] and M2 (including SNPs 19G/A, 1A/C, 27T/C and 76G/A) of were also reported to affect RPL risks. The present study aimed to investigate the association between the haplotype located in the promoter region of and the risk of RPL. Patients with RPL (n=235) or intrauterine fetus death (IUFD; n=154), as well as healthy control subjects (n=375) were enrolled in the current research. Their haplotypes of were determined using genotyping, and the association between haplotypes and the risk of RPL was accordingly analyzed. A luciferase assay was conducted to investigate the haplotype responsible for activity. Reverse transcription‑quantitative PCR, western blot analysis, immunohistochemistry and ELISA were performed to assess the expression level and activity of in patients with RPL. Consequently, the majority (n=214) of patients with RPL had a history of early RPL, whereas 31 patients with RPL had a history of both early and late RPL episodes. A significant difference was found between cases and controls in terms of gravidity and parity, whereas no significant differences were found in terms of age. The percentage of patients with RPL carrying the M2 haplotype of was significantly higher compared with that in control subjects, indicating that the M2 haplotype of was an independent risk of RPL as it influenced the transcription efficiency of promoter. In patients with RPL, ANXA5 activity was suppressed and the mRNA and protein expression levels of Anxa5 were decreased. Thus, the M2 haplotype may be an independent risk factor of RPL by affecting Anxa5 activity.
复发性妊娠丢失（RPL）常与调节异常的膜联蛋白 A5（ANXA5）表达有关。此外，富含胎盘以防止凝血的 Anxa5 蛋白的变体已被报道会影响 RPL 风险。[包括单核苷酸多态性（SNP）1A/C 和 27T/C]和 M2（包括 SNPs 19G/A、1A/C、27T/C 和 76G/A）的单倍型 M1 也被报道会影响 RPL 风险。本研究旨在探讨位于 启动子区域的单倍型与 RPL 风险之间的关系。招募了 235 例 RPL 患者和 154 例宫内胎儿死亡（IUFD）患者以及 375 例健康对照者参与本研究。使用基因分型确定他们的 单倍型，并相应分析 单倍型与 RPL 风险之间的关系。进行荧光素酶测定以研究负责 活性的单倍型。进行逆转录-定量 PCR、western blot 分析、免疫组织化学和 ELISA 以评估 RPL 患者中 ANXA5 的表达水平和活性。结果，大多数（n=214）RPL 患者有早期 RPL 病史，而 31 例 RPL 患者有早期和晚期 RPL 发作史。病例与对照组在孕次和产次方面存在显著差异，而在年龄方面无显著差异。携带 单倍型 M2 的 RPL 患者的百分比明显高于对照组，表明 单倍型 M2 是 RPL 的独立危险因素，因为它影响了 启动子的转录效率。在 RPL 患者中，ANXA5 活性受到抑制，Anxa5 的 mRNA 和蛋白表达水平降低。因此， 单倍型 M2 可能通过影响 Anxa5 活性成为 RPL 的独立危险因素。
