School of Basic Medicine, Shandong First Medical University & Shandong Academy of Medical Sciences, No. 18877 Jingshi Road, Jinan, 250062, Shandong, China.
Shandong Academy of Chinese Medicine, No. 7 West Yanzishan Road, Jinan, 250014, Shandong, China.
J Mol Med (Berl). 2022 Jul;100(7):1043-1056. doi: 10.1007/s00109-022-02178-3. Epub 2022 Jun 10.
Unexplained recurrent spontaneous abortion (URSA) is commonly observed, and seriously affects women's reproductive health. Excessive interleukin-6 (IL-6) production has been shown to frequently occur and relate to URSA pathogenesis. In this study, the miRNA expression profile in peripheral blood mononuclear cells (PBMCs) from URSA patients and normal pregnant (NP) women was assessed by miRNA microarray and real-time quantitative reverse-transcription polymerase chain reaction (qPCR). MiRNA target prediction tools and luciferase reporter assay were used to detect direct binding between miRNAs and IL6. Functional study of administering anti-IL-6 neutralizing antibody and miR-374c-5p mimics to an URSA animal model was performed to evaluate embryo resorption rates. In the results, compared with NP women, the expression of IL-6 increased markedly in PBMCs and decidual tissues at both mRNA and protein levels, while miR-374c-5p expression decreased significantly. Prediction software and luciferase reporter assay showed that miR-374c-5p binds with IL6 3'UTR via the complementary bases. Transfection of miR-374c-5p mimics into an in vitro HeLa cell line significantly downregulated the expression of IL-6, while transfection of the miR-374c-5p inhibitor induced an opposite result. In the URSA mouse model, miR-374c-5p overexpression reduced the embryo resorption rate significantly, accompanied with decreased expression of IL-6 in the decidua. To sum up, downregulated miR-374c-5p was involved in the pathogenesis of URSA by enhancing IL-6 expression. Modulation of miR-374c-5p expression may be used to regulate IL-6 production for the treatment of URSA.
不明原因复发性自然流产(URSA)较为常见,严重影响了女性的生殖健康。大量研究表明,白细胞介素 6(IL-6)过度产生与 URSA 的发病机制有关。本研究通过 miRNA 微阵列和实时定量逆转录聚合酶链反应(qPCR)评估了 URSA 患者和正常妊娠(NP)妇女外周血单个核细胞(PBMC)中的 miRNA 表达谱。利用 miRNA 靶标预测工具和荧光素酶报告基因检测,检测了 miRNA 与 IL6 之间的直接结合。通过向 URSA 动物模型中给予抗 IL-6 中和抗体和 miR-374c-5p 模拟物,进行功能研究,以评估胚胎吸收率。结果表明,与 NP 妇女相比,URSA 患者 PBMC 和蜕膜组织中 IL-6 的表达在 mRNA 和蛋白水平均显著升高,而 miR-374c-5p 的表达则显著降低。预测软件和荧光素酶报告基因检测显示,miR-374c-5p 通过互补碱基与 IL6 3'UTR 结合。将 miR-374c-5p 模拟物转染到体外 HeLa 细胞系中,显著下调了 IL-6 的表达,而转染 miR-374c-5p 抑制剂则诱导了相反的结果。在 URSA 小鼠模型中,miR-374c-5p 的过表达显著降低了胚胎吸收率,同时蜕膜中 IL-6 的表达也降低。总之,下调的 miR-374c-5p 通过增强 IL-6 的表达参与了 URSA 的发病机制。调节 miR-374c-5p 的表达可能用于调节 IL-6 的产生,以治疗 URSA。