circRNA_079813调控ROR2和ERK1/2-MAPK信号通路对牙髓损伤大鼠成骨分化的影响

周洋; 张悦; 刘景

doi:10.16190/j.cnki.45-1211/r.2023.09.004

circRNA_079813调控ROR2和ERK1/2-MAPK信号通路对牙髓损伤大鼠成骨分化的影响

Effect of circRNA_079813 promoting osteogenic differentiation via regurating ROR2 and ERK1/2-MAPK signaling pathway in dental pulp injury rats

摘要

摘要: 目的：探讨circRNA_079813对牙髓损伤模型大鼠成骨分化的作用及其机制。方法：将70只SD大鼠随机分为7组，即假手术组、牙髓损伤组、pcDNA3.1 空载体（pcDNA-null）+牙髓损伤组、过表达circRNA_079813（pcDNA-circ）+牙髓损伤组、pcDNA-circ+siRNA-NC+牙髓损伤组、pcDNA-circ+siRNA-ROR2+牙髓损伤组、pcDNA-circ+PD98059（ERK1/2 抑制剂）+牙髓损伤组，每组10 只。术后3 d 采用苏木精—伊红（HE）染色观察牙髓组织病理变化。检测各组碱性磷酸酶（ALP）活性，RTqPCR 法检测circRNA_079813 表达，western blotting 法检测ROR2、磷酸化（p-）ERK1/2、骨涎蛋白（BSP）、骨钙蛋白（OCN）、ALP、牙本质涎磷蛋白（DSPP）和牙本质基质蛋白-1（DMP-1）蛋白表达。结果：与假手术组比较，牙髓损伤组牙髓组织有明显炎症浸润和损伤特征，circRNA_079813 表达下调（P< 0.05）。与pcDNA-null+牙髓损伤组比较，pcDNA-circ+牙髓损伤组circRNA_079813、ROR2、p-ERK1/2、BSP、ALP、OCN、DSPP、DMP-1表达水平及ALP活性均升高（均P< 0.05）。沉默ROR2表达后，与pcDNA-circ+siRNA-NC+牙髓损伤组比较，pcDNA-circ+siRNA-ROR2+牙髓损伤组ROR2、p-ERK1/2、BSP、ALP、OCN、DSPP、DMP-1 表达及ALP 活性均下降（均P< 0.05）。与pcDNA-circ+牙髓损伤组比较，pcDNA-circ+PD98059+牙髓损伤组p-ERK1/2、ALP、BSP、OCN、DSPP、DMP-1表达及ALP活性均下降（均P< 0.05）。结论：circRNA_079813通过促进ROR2表达激活牙髓损伤大鼠ERK1/2-MAPK信号通路促进成骨分化。

Abstract: Objective:To explore the role and mechanism of circular RNA(circRNA)079813 in osteogenic differentiation in a dental pulp injury model in rats.Methods:A total of 70 SD rats were randomly divided into 7 groups:sham operation group, dental pulp injury group, pcDNA3.1 empty vector(pcDNA-null)+dental pulp injury group, overexpression of circRNA_079813 (pcDNA-circ)+dental pulp injury group, pcDNA-circ+siRNANC+dental pulp injury group, pcDNA-circ+siRNA-ROR2+dental pulp injury group, and pcDNA-circ+PD98059(ERK1/2 inhibitor)+dental pulp injury group, with 10 in each group.Hematoxylin-eosin(HE)staining was used to observe the pathological changes in dental pulp tissue 3 days after surgery.Alkaline phosphatase(ALP) activity was detected in each group, and circRNA_079813 expression was detected by real-time fluorescence quantitative polymerase chain reaction (RT-qPCR).The expression of ROR2, phosphorylated ERK1/2 (p-ERK1/2), bone sialoprotein (BSP), osteocalcin (OCN), ALP, dentin sialophosphoprotein (DSPP) and dentin matrix protein-1(DMP-1)were detected by western blotting.Results:Compared with the sham operation group, the dental pulp tissue in the dental pulp injury group showed significant inflammatory infiltration and damage characteristics, and the expression of circRNA_079813 was down-regulated (P< 0.05).Compared with the pcDNA-null+dental pulp injury group, the expression levels of circRNA_079813, ROR2, p-ERK1/2, BSP, ALP, OCN, DSPP and DMP-1 and the activity of ALP were up-regulated in the pcDNA-circ+dental pulp injury group(all P< 0.05).After silencing ROR2 expression, compared with the pcDNA-circ+siRNA-NC+dental pulp injury group, the expression of ROR2, p-ERK1/2, BSP, ALP, OCN, DSPP and DMP-1 and the activity of ALP in the pcDNA-circ+siRNA-ROR2+dental pulp injury group were down-regulated(all P< 0.05).Compared with the pcDNA-circ+dental pulp injury group, the expression of p-ERK1/2, ALP, BSP, OCN, DSPP and DMP-1 and the activity of ALP were down-regulated in the pcDNA-circ+PD98059+dental pulp injury group(all P< 0.05).Conclusion:CircRNA_079813 promotes osteogenic differentiation in dental pulp injury rats through activation of the ERK1/2-MAPK signaling pathway by promoting ROR2 expression.

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