Jing Wang1, Chenchen Du2, Lulu Xu3. 1. Department of Orthodontics, Chinese PLA General Hospital, Beijing, 100853, China. 2. Department of Stomatology, Xinjiang Karamay People's Hospital, Karamay, Xinjiang, 834000, China. 3. Department of Orthodontics, Chinese PLA General Hospital, Beijing, 100853, China. Electronic address: elsovu@163.com.
Abstract
OBJECTIVES: Revealing the role and mechanism of circ_0081572 in periodontitis progression. DESIGN: 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. RESULTS: 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. CONCLUSIONS: Our results suggested that circ_0081572 might prevent periodontitis by regulating the miR-378h /RORA axis.
OBJECTIVES: Revealing the role and mechanism of circ_0081572 in periodontitis progression. DESIGN: 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. RESULTS: Circ_0081572 was a stability circRNA with downregulated expression in the gingival tissues of periodontitispatients. 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. CONCLUSIONS: Our results suggested that circ_0081572 might prevent periodontitis by regulating the miR-378h /RORA axis.