miRNA-624-5p 在先天性甲状腺功能减退症中的作用及其通过靶向 SIRT1 的分子机制。

The role of miRNA-624-5p in congenital hypothyroidism and its molecular mechanism by targeting SIRT1.

机构信息

Department of Pediatrics, Yiwu Maternity and Children Hospital, No. C100 Xinke Road, Yiwu, 322000, China.

出版信息

Genes Genomics. 2022 Sep;44(9):1137-1147. doi: 10.1007/s13258-021-01171-3. Epub 2021 Oct 5.

DOI:10.1007/s13258-021-01171-3

PMID:34609722

Abstract

BACKGROUND

Accumulating reports evidenced that congenital hypothyroidism (CH) is a kind of endocrine diseases caused by thyroid hormone imperfection. MicroRNAs (miRNAs) were confirmed to exhibit protective functions in CH progression. However, the functions and latent mechanism of microRNA-624-5p (miR-624-5p) in CH remains unknown.

OBJECTIVE

This report was designed to illustrate the potential molecular mechanisms of miR-624-5p on CH.

METHODS

Rats were induced by 50 mg/day propylthiouracil to conduct CH models. Free thyroxine (fT4) and thyroid-Stimulating hormone (TSH) concentrations were measured to confirm CH model conduction. The direct target of miR-624-5p was predicted and verified by Starbase and dual luciferase reporter assay. Besides, the levels of miR-624-5p and sirtuin1 (SIRT1) in hippocampus or hippocampal neuronal cells were assessed using quantitative real-time polymerase chain reaction (qRT-PCR) or Western blot assays. Then CH rat behaviors were evaluated using open field test (OFT) and forced swim test (FST). Furthermore, neuronal cells viability and apoptosis were checked using 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) and flow cytometry.

RESULTS

qRT-PCR assay suggested that miR-624-5p was up-regulated and SIRT1 was low-expressed in hippocampus tissues of CH rats. SIRT1 was a direct target of miR-624-5p. MiR-624-5p inhibitor signally enhanced fT4 levels and reduced TSH levels in the plasma of CH rats, and improved CH rat depressive behaviors by targeting SIRT1. Moreover, our data also revealed that miR-624-5p inhibitor increased cell viability and reduced apoptotic neuronal cells, which was reversed by silencing of SIRT1.

CONCLUSIONS

Taken together, this research demonstrated that miR-624-5p serves as a promising target for CH treatment.

摘要

背景

越来越多的报道表明，先天性甲状腺功能减退症（CH）是一种由甲状腺激素缺陷引起的内分泌疾病。微小 RNA（miRNA）已被证实对 CH 的进展具有保护作用。然而，miR-624-5p 在 CH 中的功能和潜在机制尚不清楚。

目的

本报告旨在阐述 miR-624-5p 对 CH 的潜在分子机制。

方法

用 50mg/天丙硫氧嘧啶诱导大鼠制作 CH 模型。测定游离甲状腺素（fT4）和促甲状腺激素（TSH）浓度，以确认 CH 模型的制作。通过 Starbase 和双荧光素酶报告基因实验预测和验证 miR-624-5p 的直接靶标。此外，采用实时定量聚合酶链反应（qRT-PCR）或 Western blot 法检测海马或海马神经元细胞中 miR-624-5p 和 SIRT1 的水平。然后采用旷场试验（OFT）和强迫游泳试验（FST）评估 CH 大鼠的行为。进一步采用 3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2-H-四唑溴盐（MTT）和流式细胞术检测神经元细胞的活力和凋亡情况。

结果

qRT-PCR 检测显示，CH 大鼠海马组织中 miR-624-5p 呈上调表达，SIRT1 呈低表达。SIRT1 是 miR-624-5p 的直接靶标。MiR-624-5p 抑制剂可显著提高 CH 大鼠血浆中的 fT4 水平，降低 TSH 水平，并通过靶向 SIRT1 改善 CH 大鼠的抑郁行为。此外，我们的数据还表明，miR-624-5p 抑制剂可增加细胞活力，减少凋亡的神经元细胞，而沉默 SIRT1 可逆转这一作用。