人羊膜间充质干细胞通过 ANRIL/miR-125a/APC 轴促进脂多糖诱导的人骨髓间充质干细胞成骨分化。
Human amnion-derived mesenchymal stem cells promote osteogenic differentiation of lipopolysaccharide-induced human bone marrow mesenchymal stem cells via ANRIL/miR-125a/APC axis.
机构信息
Jiangsu Key Laboratory of Oral Diseases, Nanjing Medical University, Nanjing, China.
Department of Oral and Maxillofacial Surgery, Affiliated Hospital of Stomatology, Nanjing Medical University, Nanjing, China.
出版信息
Stem Cell Res Ther. 2021 Jan 7;12(1):35. doi: 10.1186/s13287-020-02105-8.
BACKGROUND AND AIM
Periodontitis is a chronic inflammatory disease inducing the absorption of alveolar bone and leading to tooth loss. Human amnion-derived mesenchymal stem cells (HAMSCs) have been used for studying inflammatory processes. This study aimed to explore the role of long noncoding RNA (lncRNA) antisense noncoding RNA in the INK4 locus (ANRIL) in HAMSC-driven osteogenesis in lipopolysaccharide (LPS)-induced human bone marrow mesenchymal stem cells (HBMSCs).
METHODS
The cells were incubated with a co-culture system. Reactive oxygen species (ROS) level and superoxide dismutase (SOD) activity were used to detect the oxidative stress level. Flow cytometry was performed to determine cell proliferation. The alkaline phosphatase (ALP) activity, Alizarin red assay, cell transfection, and rat mandibular defect model were used to evaluate the osteogenic differentiation. Quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR), Western blot analysis, dual-luciferase reporter assay, and immunofluorescence staining were used to evaluate the molecular mechanisms.
RESULTS
This study showed that HAMSCs promoted the osteogenesis of LPS-induced HBMSCs, while the ANRIL level in HBMSCs decreased during co-culture. ANRIL had no significant influence on the proliferation of LPS-induced HBMSCs. However, its overexpression inhibited the HAMSC-driven osteogenesis in vivo and in vitro, whereas its knockdown reversed these effects. Mechanistically, this study found that downregulating ANRIL led to the overexpression of microRNA-125a (miR-125a), and further contributed to the competitive binding of miR-125a and adenomatous polyposis coli (APC), thus significantly activating the Wnt/β-catenin pathway.
CONCLUSION
The study indicated that HAMSCs promoted the osteogenic differentiation of LPS-induced HBMSCs via the ANRIL/miR-125a/APC axis, and offered a novel approach for periodontitis therapy.
背景与目的
牙周炎是一种慢性炎症性疾病，会导致牙槽骨吸收，进而导致牙齿脱落。人羊膜间充质干细胞（HAMSCs）已被用于研究炎症过程。本研究旨在探讨长链非编码 RNA（lncRNA）反义非编码 RNA 在 INK4 基因座（ANRIL）在脂多糖（LPS）诱导的人骨髓间充质干细胞（HBMSCs）中 HAMSC 驱动的成骨中的作用。
方法
将细胞与共培养系统孵育。用活性氧（ROS）水平和超氧化物歧化酶（SOD）活性来检测氧化应激水平。用流式细胞术检测细胞增殖。用碱性磷酸酶（ALP）活性、茜素红染色、细胞转染和大鼠下颌骨缺损模型来评估成骨分化。用实时定量逆转录-聚合酶链反应（RT-PCR）、Western blot 分析、双荧光素酶报告基因检测和免疫荧光染色来评估分子机制。
结果
本研究表明 HAMSCs 促进了 LPS 诱导的 HBMSCs 的成骨作用，而在共培养过程中 HBMSCs 中的 ANRIL 水平降低。ANRIL 对 LPS 诱导的 HBMSCs 的增殖没有显著影响。然而，其过表达抑制了体内和体外的 HAMSC 驱动的成骨作用，而其敲低则逆转了这些作用。机制上，本研究发现下调 ANRIL 导致 microRNA-125a（miR-125a）的过表达，并进一步导致 APC 的竞争结合，从而显著激活 Wnt/β-catenin 通路。
结论
该研究表明 HAMSCs 通过 ANRIL/miR-125a/APC 轴促进 LPS 诱导的 HBMSCs 的成骨分化，为牙周炎的治疗提供了一种新方法。
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