Department of Stomatology, Huaian Maternal and Child Health Hospital, Huaian 223300, Jiangsu, China.
Department of Stomatology, The Affiliated Huaian No. 1 People's Hospital of Nanjing Medical University, Huaian 223300, Jiangsu, China.
Arch Oral Biol. 2022 Jan;133:105298. doi: 10.1016/j.archoralbio.2021.105298. Epub 2021 Oct 29.
This study aims to investigate the role of long noncoding RNA distal-less homeobox 2 antisense 1 (DLX2-AS1) in lipopolysaccharide-induced inflammatory response and apoptosis of periodontal ligament cells (PDLCs).
Lipopolysaccharide was used to induce inflammation response of PDLCs. The expression of DLX2-AS1, microRNA-330-3p and Ro60, Y RNA binding protein (RO60) in lipopolysaccharide-treated PDLCs was detected by reverse transcription quantitative polymerase chain reaction (RT-qPCR). Enzyme linked immunosorbent assay (ELISA) was performed to evaluate the concentration of inflammatory cytokines in PDLCs after DLX2-AS1 overexpression or RO60 downregulation. The apoptosis of PDLCs after lipopolysaccharide treatment or indicated transfection was analyzed by flow cytometry analysis. The level of apoptosis-related proteins, Bax and Bcl-2, were examined by western blotting. The binding capacity between microRNA-330-3p and DLX2-AS1 (or RO60) was verified by luciferase reporter assays.
DLX2-AS1 was downregulated in PDLCs after lipopolysaccharide treatment. DLX2-AS1 overexpression decreased the production of inflammatory cytokines and inhibited cell apoptosis. microRNA-330-3p bound with DLX2-AS1 and displayed high expression in lipopolysaccharide-induced PDLCs. In addition, the downregulation of RO60, a target gene of microRNA-330-3p, reversed the suppressive influence of DLX2-AS1 overexpression on the inflammatory response and apoptosis of PDLCs.
DLX2-AS1 restrains inflammatory response and apoptosis of PDLCs via the microRNA-330-3p/RO60 axis.
本研究旨在探讨长链非编码 RNA 远隔同源盒 2 反义 1（DLX2-AS1）在脂多糖诱导的牙周膜细胞（PDLCs）炎症反应和细胞凋亡中的作用。
用脂多糖诱导 PDLCs 炎症反应，采用反转录定量聚合酶链反应（RT-qPCR）检测脂多糖处理的 PDLCs 中 DLX2-AS1、microRNA-330-3p 和 Ro60，Y 核酸结合蛋白（RO60）的表达。采用酶联免疫吸附试验（ELISA）检测 DLX2-AS1 过表达或 RO60 下调后 PDLCs 中炎症细胞因子的浓度。用流式细胞术分析脂多糖处理或转染后 PDLCs 的凋亡情况。用蛋白质印迹法检测凋亡相关蛋白 Bax 和 Bcl-2 的水平。通过荧光素酶报告基因实验验证 microRNA-330-3p 与 DLX2-AS1（或 RO60）的结合能力。
脂多糖处理后 PDLCs 中 DLX2-AS1 下调。DLX2-AS1 过表达可降低炎症细胞因子的产生并抑制细胞凋亡。microRNA-330-3p 与 DLX2-AS1 结合，在脂多糖诱导的 PDLCs 中表达水平较高。此外，microRNA-330-3p 靶基因 RO60 的下调逆转了 DLX2-AS1 过表达对 PDLCs 炎症反应和凋亡的抑制作用。
DLX2-AS1 通过 microRNA-330-3p/RO60 轴抑制 PDLCs 的炎症反应和凋亡。
