褪黑素与 circ_0005753/PTBP1/TXNIP 调控网络在调节间充质干细胞成骨潜能中的新联系。

A novel link between melatonin and circ_0005753/PTBP1/TXNIP regulatory network in the modulation of osteogenic potential in mesenchymal stem cells.

机构信息

Endocrine Department, The Second Affiliated Hospital of Nanchang University, Nanchang, Jiangxi Province, China.

Department of Nephrology, The Second Affiliated Hospital of Nanchang University, Nanchang, Jiangxi Province, China.

出版信息

Chem Biol Drug Des. 2024 Jan;103(1):e14380. doi: 10.1111/cbdd.14380. Epub 2023 Oct 27.

DOI:10.1111/cbdd.14380

PMID:37890873

Abstract

Labeled with pluripotent potential, the transplantation of bone marrow mesenchymal stem cells (BMSCs) is considered as a promising strategy for treating osteoporosis (OP). Melatonin (MEL) has been investigated to be an essential regulator involved in bone metabolism, as well as BMSCs differentiation. Circular RNAs (circRNAs) are a unique kind of non-coding RNA and play an important regulatory role in OP. However, whether circRNAs are implicated in the effects of MEL on BMSCs osteogenic differentiation remains largely indeterminate. Expression of circ_0005753 in human BMSCs with MEL treatment, clinical specimens diagnosed with OP, either with ovariectomy (OVX)-induced mice, was measured by RT-qPCR. Western blot was conducted to analyze protein levels of osteogenesis-related molecules (Opg, RUNX2, ALP, BMP4) and TXNIP. RNA immunoprecipitation (RIP) and RNA pull-down assays were performed to validate the binding relationship among circ_0005753, PTBP1, and TXNIP. Alkaline phosphatase (ALP) and alizarin red staining (ARS) were performed to evaluate osteogenic capacity of BMSCs. OP mouse model was established by ovariectomy, as evaluated pathologic changes via hematoxylin-eosin (HE), Masson, and Immunohistochemistry (IHC) staining. Expression of circ_0005753 was remarkably decreased during MEL-induced osteogenic differentiation of BMSCs. Interestingly, not only circ_0005753 knockdown significantly promoted osteogenic differentiation of BMSCs, but circ_0005753 overexpression also weakened osteogenic differentiation induced by MEL treatment. Mechanistically, circ_0005753 maintained the stabilization of TXNIP mRNA via recruiting PTBP1. Additionally, reinforced circ_0005753 abrogated MEL-mediated protective effects on OP pathogenesis in a mouse model. This work shows that MEL facilitates osteogenic differentiation of BMSCs via the circ_0005753/PTBP1/TXNIP axis, which may shed light on the development of a novel therapeutic strategy to prevent OP.

摘要

用多潜能性标记的骨髓间充质干细胞（BMSCs）移植被认为是治疗骨质疏松症（OP）的有前途的策略。褪黑素（MEL）已被研究为参与骨代谢以及 BMSCs 分化的重要调节剂。环状 RNA（circRNA）是一种独特的非编码 RNA，在 OP 中发挥重要的调节作用。然而，circRNA 是否参与 MEL 对 BMSCs 成骨分化的影响仍在很大程度上不确定。用 MEL 处理人 BMSCs 后，通过 RT-qPCR 测量 circ_0005753 的表达，用 RT-qPCR 测量临床诊断为 OP 的标本中 circ_0005753 的表达，或者用卵巢切除术（OVX）诱导的小鼠。通过 Western blot 分析成骨相关分子（Opg、RUNX2、ALP、BMP4）和 TXNIP 的蛋白水平。进行 RNA 免疫沉淀（RIP）和 RNA 下拉测定以验证 circ_0005753、PTBP1 和 TXNIP 之间的结合关系。进行碱性磷酸酶（ALP）和茜素红染色（ARS）以评估 BMSCs 的成骨能力。通过卵巢切除术建立 OP 小鼠模型，通过苏木精-伊红（HE）、Masson 和免疫组织化学（IHC）染色评估病理变化。在 MEL 诱导的 BMSCs 成骨分化过程中，circ_0005753 的表达明显降低。有趣的是，不仅 circ_0005753 敲低显着促进 BMSCs 的成骨分化，而且 circ_0005753 的过表达也削弱了 MEL 处理诱导的成骨分化。机制上，circ_0005753 通过募集 PTBP1 来维持 TXNIP mRNA 的稳定性。此外，增强的 circ_0005753 消除了 MEL 对 OP 发病机制的保护作用在小鼠模型中。这项工作表明，MEL 通过 circ_0005753/PTBP1/TXNIP 轴促进 BMSCs 的成骨分化，这可能为开发预防 OP 的新治疗策略提供启示。