Nong Dongmei, Qin Yaqing, Zhou Hua, Kang Na
Guangxi Key Laboratory of Oral and Maxillofacial Rehabilitation and Reconstruction, Guangxi Clinical Research Center for Craniofacial Deformity, Guangxi Key Laboratory of Oral and Maxillofacial Surgery Disease Treatment, Guangxi Medical University, Nanning 530021, China.
Guangxi Key Laboratory of Oral and Maxillofacial Rehabilitation and Reconstruction, Guangxi Clinical Research Center for Craniofacial Deformity, Guangxi Key Laboratory of Oral and Maxillofacial Surgery Disease Treatment, Guangxi Medical University, Nanning 530021, China.*Corresponding author, E-mail:
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2019 Jun;35(6):545-551.
Objective To illuminate whether IL-17 regulates receptor activator of NF-κB ligand (RANKL) and osteoprotegerin (OPG) expression in human periodontal ligament fibroblasts (HPDLFs) via p38MAPK signaling pathway. Methods HPDLFs were incubated in the presence of 20 ng/mL IL-17 for 0, 20, 40, 60 and 80 minutes. HPDLFs were divided randomly into 6 groups: control group, dimethyl sulfoxide (DMSO) group, p38MAPK pathway inhibitor SB203580 group, IL-17 group, IL-17 combined with DMSO group and IL-17 combined with SB203580 group. SB203580 (10 μmol/L) and IL-17 (20 ng/mL) were added to the corresponding groups. Real-time quantitative PCR was used to detect the expression of RANKL and OPG mRNAs in HPDLFs. The levels of phospho-p38MAPK (p-p38MAPK) and RANKL protein were measured using Western blot analysis. The protein level of OPG was detected by ELISA. Results After IL-17 stimulation, the expression level of p-p38MAPK protein gradually increased starting from 0 minute and reached its highest level at 60 minutes. It started to decline at 80 minutes. Stimulation with IL-17 could increase the mRNA and protein expression level of RANKL but decrease the mRNA and protein expression level of OPG. Nevertheless, unlike the IL-17 group, IL-17 combined with inhibitor SB203580 decreased the expression of RANKL mRNA and protein and increased OPG mRNA. Conclusion IL-17 can enhance the expression of RANKL in human periodontal fibroblasts and inhibit the expression of OPG mRNA through the p38MAPK signal transduction pathways.
目的 探讨白细胞介素-17（IL-17）是否通过p38丝裂原活化蛋白激酶（p38MAPK）信号通路调控人牙周膜成纤维细胞（HPDLFs）中核因子κB受体活化因子配体（RANKL）和骨保护素（OPG）的表达。方法 将HPDLFs分别在20 ng/mL IL-17作用0、20、40、60和80分钟。HPDLFs随机分为6组：对照组、二甲基亚砜（DMSO）组、p38MAPK通路抑制剂SB203580组、IL-17组、IL-17联合DMSO组和IL-17联合SB203580组。相应组分别加入SB203580（10 μmol/L）和IL-17（20 ng/mL）。采用实时定量聚合酶链反应（PCR）检测HPDLFs中RANKL和OPG mRNA的表达。采用蛋白质免疫印迹法检测磷酸化p38MAPK（p-p38MAPK）和RANKL蛋白水平。采用酶联免疫吸附测定法（ELISA）检测OPG蛋白水平。结果 IL-17刺激后，p-p38MAPK蛋白表达水平从0分钟开始逐渐升高，60分钟时达到最高水平，80分钟时开始下降。IL-17刺激可增加RANKL的mRNA和蛋白表达水平，但降低OPG的mRNA和蛋白表达水平。然而，与IL-17组不同，IL-17联合抑制剂SB203580可降低RANKL mRNA和蛋白的表达，并增加OPG mRNA的表达。结论 IL-17可通过p38MAPK信号转导通路增强人牙周成纤维细胞中RANKL的表达，并抑制OPG mRNA的表达。
