摘要
目的:研究miR-140-5p对脂多糖(lipopolysaccharides,LPS)诱导牙髓细胞炎症因子分泌的影响和机制。方法:分离培养人牙髓细胞,分为对照组、LPS组、miR-NC+LPS组、miR-140-5p+LPS组,Realtime PCR检测细胞中miR-140-5p表达水平,ELISA法检测细胞培养上清液中IL-6、IL-1β和TNF-α含量,Western Blot检测细胞中p-IκBα、IκBα、NF-κBp65蛋白表达水平。用NF-κB信号通路激活剂处理LPS条件下转染miR-140-5p mimics的人牙髓细胞,ELISA法检测培养液上清中IL-6、IL-1β、TNF-α含量,Western Blot检测细胞中p-IκBα、IκBα、NF-κBp65蛋白表达。结果:与对照组比较,LPS组细胞中miR-140-5p水平下降,细胞培养液上清中IL-6、IL-1β、TNF-α含量升高,细胞中p-IκBα、NF-κBp65蛋白表达增多,IκBα蛋白表达没有变化。与miR-NC+LPS组比较,miR-140-5p+LPS组细胞中miR-140-5p水平升高,细胞培养液上清中IL-6、IL-1β、TNF-α含量降低,细胞中p-IκBα、NF-κBp65蛋白表达减少,IκBα蛋白表达没有变化。与未经NF-κB信号通路激活剂处理的细胞比较,NF-κB信号通路激活剂可以促进上调miR-140-5p表达的LPS条件下牙髓细胞中p-IκBα、NF-κBp65蛋白表达和IL-6、IL-1β、TNF-α分泌。结论:miR-140-5p通过调控NF-κB信号通路降低LPS诱导的人牙髓细胞炎症因子的分泌。
Objective:To investigate the effect and mechanism of miR-140-5 p on secretion of inflammatory cytokines in lipopolysaccharides(LPS)-induced dental pulp cells.Methods:Human dental pulp cells were isolated and cultured,then they were divided into control group,LPS group,miR-NC+LPS group,miR-140-5 p+LPS group.Realtime PCR was used to detect the expression level of miR-140-5 p in cells.ELISA was used to detect the levels of IL-6,IL-1βand TNF-αin the cell culture supernatant.Western Blot was used to detect the levels of p-IκBα,IκBα,NF-κBp65 protein in cells.Human dental pulp cells transfected with miR-140-5 p mimics were treated with NF-κB signal activator,then ELISA was used to detect the levels of IL-6,IL-1βand TNF-αin the cell culture supernatant,and Western Blot was used to detect the levels of p-IκBα,IκBα,NF-κBp65 protein in cells.Results:Compared with the control group,the level of miR-140-5 p in the LPS group decreased,the levels of IL-6,IL-1βand TNF-αin the cell culture supernatant increased,and the expression levels of p-IκBαand NF-κBp65 protein in cells increased,there was no change in the expression of IκBαprotein.Compared with the miR-NC+LPS group,the level of miR-140-5 p was increased in the miR-140-5 p+LPS group,and the levels of IL-6,IL-1βand TNF-αin the culture supernatant were decreased,and the expression levels of p-IκBαand NF-κBp65 protein in cells reduced,there was no change in the expression of IκBαprotein.Compared with the cells not treated with NF-κB signal pathway activator,NF-κB signal pathway activator could promote the expression of p-IκBα,NF-κBp65 protein and the secretion of IL-6,IL-1β,TNF-αin the pulp cells under the LPS condition of up regulating the expression of miR-140-5 p.Conclusion:miR-140-5 p reduces the secretion of inflammatory factors in LPS-induced dental pulp cells by regulating NF-κB signaling pathway.
作者
黄伊娜
常圣
刘从勇
HUANG Yi-na;CHANG Sheng;LIU Cong-yong(Department of Stomatology,Chengdu Public Health Clinical Center,Sichuan Chengdu 060000,China)
出处
《临床口腔医学杂志》
2020年第12期716-720,共5页
Journal of Clinical Stomatology
