Department of Endocrinology and Metabolism, Institute of Endocrinology, Liaoning, Provincial Key Laboratory of Endocrine Diseases, The First Affiliated Hospital of China Medical University, Shenyang, Liaoning, China (mainland).
Shijiazhuang First Hospital, Shijiazhuang, Hebei, China (mainland).
Med Sci Monit. 2018 Jul 20;24:5041-5049. doi: 10.12659/MSM.907510.
BACKGROUND A deficiency of maternal thyroid hormones (THs) during pregnancy has severe impacts on fetal brain development. Neural stem cells (NSCs) are major targets of THs and provided a powerful model to explore the underlying mechanism of THs during brain development. Although miRNA-125 might be associated with the NSCs differentiation, the relationship between miR-125 and hypothyroidism (HypoT) development remains unclear. MATERIAL AND METHODS In our study, we screened a differentially expressed gene miR-125b-5p from brain between euthyroid (EuT) and HypoT rats. In vitro, we employed anion exchange resin to remove THs to stimulate HypoT. QRT-PCR and Western blot were used to examine the expression of signal transducer and activator of transcription 3 (Stat3). The relationship between miR-125b-5p and Stat3 was detected via a dual-luciferase assay. RESULTS QRT-PCR results showed that the level of miR-125b-5p in HypoT rat brains was significantly suppressed, suggesting some relationship between miR-125b-5p and HypoT. In C17.2, miR-125b-5p promoted cell differentiation into neurons by regulating the expression of tubulin beta chain 3 (TUBB3) and glial fibrillary acid protein (GFAP). QRT-PCR and Western blot results revealed that miR-125b-5p mimic modulated the contents of total Stat3 and p-Stat3. A dual-luciferase assay showed that miR-125b-5p negatively regulated the expression of Stat3 by binding with the first site in 3' UTR of Stat3. CONCLUSIONS These results revealed Stat3 is a new target of miR-125b-5p and revealed the mechanism of miR-125b-5p suppressing HypoT development. These findings provide a new target for HypoT therapy.
妊娠期间母体甲状腺激素(THs)缺乏对胎儿大脑发育有严重影响。神经干细胞(NSCs)是 THs 的主要靶标,并为探索 THs 在大脑发育过程中的潜在机制提供了一个强大的模型。虽然 miRNA-125 可能与 NSCs 分化有关,但 miR-125 与甲状腺功能减退症(HypoT)发展之间的关系尚不清楚。
在我们的研究中,我们从正常甲状腺功能(EuT)和 HypoT 大鼠的大脑中筛选出差异表达的基因 miR-125b-5p。在体外,我们采用阴离子交换树脂去除 THs 来刺激 HypoT。采用 QRT-PCR 和 Western blot 检测信号转导和转录激活因子 3(Stat3)的表达。通过双荧光素酶报告基因检测 miR-125b-5p 与 Stat3 之间的关系。
QRT-PCR 结果显示 HypoT 大鼠大脑中的 miR-125b-5p 水平明显受到抑制,提示 miR-125b-5p 与 HypoT 之间存在一定的关系。在 C17.2 中,miR-125b-5p 通过调节微管相关蛋白β链 3(TUBB3)和神经胶质纤维酸性蛋白(GFAP)的表达促进细胞向神经元分化。QRT-PCR 和 Western blot 结果显示,miR-125b-5p 模拟物调节总 Stat3 和 p-Stat3 的含量。双荧光素酶报告基因显示,miR-125b-5p 通过结合 Stat3 3'UTR 的第一位点负调控 Stat3 的表达。
这些结果表明 Stat3 是 miR-125b-5p 的一个新靶点,并揭示了 miR-125b-5p 抑制 HypoT 发展的机制。这些发现为 HypoT 治疗提供了一个新的靶点。