IADR Abstract Archives
EZH2 Plays a Role in Extracellular Matrix Degradation of Pulpitis
Objectives: Dental pulp inflammation is a prerequisite for pulp healing, with series of events ahead of regeneration, including extracellular matrix (ECM) degradation. The aim of this study was to explore the effect of the epigenetic mark, enhancer of zeste homolog 2 (EZH2) on ECM during pulpitis and to identify the underlying mechanisms in human dental pulp cells (HDPCs).
Methods: The effects of EZH2 and EZH2 inhibitor (EI1) on matrix metalloproteinases (MMPs) and type I collagen (Collagen I) in HDPCs were evaluated by quantitative polymerase chain reaction (qPCR). The protein level of Collagen I was examined by western blot analysis. A rat model of dental pulp inflammation was established, and the expression of Collagen I in dental pulp under EZH2 or EI1 stimulation was detected by immunohistochemical staining. We applied miRcode data combined with qPCR to screen candidate miRNAs mediating the regulation of MMPs by EZH2. The effects of the mimic and inhibitor of the miRNA on ECM has been detected by qPCR and western blot.
Results: EZH2 induced the expression of MMPs (MMP-1, MMP-3, MMP-8 and MMP-10) and reduced expression of Collagen I. The EI1 treatment downregulated the expression of ECM molecules, such as MMP-3, MMP-2 and MMP-8, while upregulated the gene expression of Collagen I but not its protein level. The expression of Collagen I has not been induced in infected pulp tissue treated by EI1. We found that miR-181 has been upregulated obviously by EI1. MiR-181 inhibition induced the gene expression and protein level of Collagen I in HDPCs. The production of ECM molecules, such as TGF-β2, FN1, ITGA5 and FNDC5 has been upregulated by miR-181 inhibitor.
Conclusions: EZH2 might affect ECM homeostasis in dental pulp inflammation. Inhibition of EZH2 and miR-181 is potent components of reducing ECM degradation in HDPCs and may play important roles in pulpitis.
Methods: The effects of EZH2 and EZH2 inhibitor (EI1) on matrix metalloproteinases (MMPs) and type I collagen (Collagen I) in HDPCs were evaluated by quantitative polymerase chain reaction (qPCR). The protein level of Collagen I was examined by western blot analysis. A rat model of dental pulp inflammation was established, and the expression of Collagen I in dental pulp under EZH2 or EI1 stimulation was detected by immunohistochemical staining. We applied miRcode data combined with qPCR to screen candidate miRNAs mediating the regulation of MMPs by EZH2. The effects of the mimic and inhibitor of the miRNA on ECM has been detected by qPCR and western blot.
Results: EZH2 induced the expression of MMPs (MMP-1, MMP-3, MMP-8 and MMP-10) and reduced expression of Collagen I. The EI1 treatment downregulated the expression of ECM molecules, such as MMP-3, MMP-2 and MMP-8, while upregulated the gene expression of Collagen I but not its protein level. The expression of Collagen I has not been induced in infected pulp tissue treated by EI1. We found that miR-181 has been upregulated obviously by EI1. MiR-181 inhibition induced the gene expression and protein level of Collagen I in HDPCs. The production of ECM molecules, such as TGF-β2, FN1, ITGA5 and FNDC5 has been upregulated by miR-181 inhibitor.
Conclusions: EZH2 might affect ECM homeostasis in dental pulp inflammation. Inhibition of EZH2 and miR-181 is potent components of reducing ECM degradation in HDPCs and may play important roles in pulpitis.