Department of Orthodontics, Chinese PLA General Hospital, Beijing, 100853, China.
Department of Stomatology, Xinjiang Karamay People's Hospital, Karamay, Xinjiang, 834000, China.
Arch Oral Biol. 2021 Apr;124:105053. doi: 10.1016/j.archoralbio.2021.105053. Epub 2021 Jan 23.
Revealing the role and mechanism of circ_0081572 in periodontitis progression.
Quantitative real-time PCR (qRT-PCR) was applied to measure the expression of circ_0081572, microRNA (miR)-378h and retinoid acid-related orphan receptor A (RORA). Lipopolysaccharide (LPS) was used to treat periodontal ligament cells (PDLCs) to construct periodontitis cell model in vitro. Cell counting kit 8 (CCK8) assay and flow cytometry were used to measure cell viability and apoptosis. The caspase 3 activity was detected by Caspase 3 Activity Assay Kit. Western blot assay was performed to detect the expression of apoptosis-associated proteins and RORA. The inflammation response and oxidative stress were determined by detecting the levels of inflammatory cytokines and reactive oxygen species (ROS). The relationship between miR-378h and circ_0081572 or RORA was verified by dual-luciferase reporter assay, RNA immunoprecipitation (RIP) assay and biotin-labeled RNA pull-down assay.
Circ_0081572 was a stability circRNA with downregulated expression in the gingival tissues of periodontitis patients. Overexpression of circ_0081572 could alleviate LPS-induced PDLCs injury. Circ_0081572 could serve as a sponge for miR-378h. Furthermore, miR-378h could reverse the inhibition of circ_0081572 on LPS-induced PDLCs injury. In addition, RORA could be targeted by miR-378h, and its silencing could reverse the suppressive effect of miR-378h inhibitor and circ_0081572 overexpression on LPS-induced PDLCs injury.
Our results suggested that circ_0081572 might prevent periodontitis by regulating the miR-378h /RORA axis.
揭示 circ_0081572 在牙周炎进展中的作用和机制。
采用实时定量 PCR (qRT-PCR)检测 circ_0081572、微小 RNA (miR)-378h 和维甲酸相关孤儿受体 A (RORA)的表达。采用脂多糖 (LPS)处理牙周膜细胞 (PDLCs)构建体外牙周炎细胞模型。细胞计数试剂盒 8 (CCK8) 检测和流式细胞术检测细胞活力和凋亡。Caspase 3 活性检测试剂盒检测 caspase 3 活性。Western blot 检测凋亡相关蛋白和 RORA 的表达。通过检测炎症细胞因子和活性氧 (ROS)水平来确定炎症反应和氧化应激。通过双荧光素酶报告基因检测、RNA 免疫沉淀 (RIP) 检测和生物素标记的 RNA 下拉实验验证 miR-378h 与 circ_0081572 或 RORA 的关系。
circ_0081572 是一种具有下调表达的稳定 circRNA，存在于牙周炎患者的牙龈组织中。circ_0081572 的过表达可减轻 LPS 诱导的 PDLCs 损伤。circ_0081572 可以作为 miR-378h 的海绵。此外，miR-378h 可以逆转 circ_0081572 对 LPS 诱导的 PDLCs 损伤的抑制作用。另外，RORA 可以作为 miR-378h 的靶基因，其沉默可以逆转 miR-378h 抑制剂和 circ_0081572 过表达对 LPS 诱导的 PDLCs 损伤的抑制作用。
我们的结果表明，circ_0081572 可能通过调节 miR-378h/RORA 轴来预防牙周炎。
