lncRNA TP53TG1在牙髓中的表达和对牙髓干细胞转录炎症因子的影响
Expression of lncRNA TP53TG1 in dental pulp and its effect on transcription of inflammatory factors in dental pulp stem cells
代子寒 1王明浩 1王胜朝 1何文喜2
作者信息
- 1. 口颌系统重建与再生全国重点实验室,国家口腔疾病临床医学研究中心,陕西省口腔医学重点实验室,空军军医大学口腔医院牙体牙髓病科,陕西西安 710032
- 2. 空军特色医学中心口腔科,北京 100142
- 折叠
摘要
目的 探究lncRNA TP53TG1在健康和炎症牙髓中的表达差异,以及对脂多糖(LPS)诱导后人牙髓干细胞(hDPSCs)炎症因子转录水平的影响.方法 设计合成lncRNA探针,利用RNA荧光原位杂交技术检测健康和炎症牙髓组织中lncRNA TP53TG1的表达情况.通过酶解组织块法分离培养hDPSCs,对培养的hDPSCs进行多向分化诱导验证,并通过流式细胞术鉴定表型特征.将hDPSCs分为si-NC组、si-NC+LPS组、si-TP53TG1+LPS组,分别转染对照序列和TP53TG1的siRNA,再用LPS处理si-NC+LPS组和si-TP53TG1+LPS组24 h,通过qRT-PCR检测各组炎症因子IL-1β、IL-8以及TNF-α的mRNA表达水平.结果 在健康牙髓组织中可观察到lncRNA TP53TG1的表达,主要分布在成牙本质细胞中.炎症状态下,lncRNA TP53TG1的表达量上升,全牙髓可见分布.LPS处理后,hDPSC的TNF-α、IL-1β以及IL-8的mRNA表达水平上升,下调lncRNA TP53TG1导致hDPSCs的TNF-α mRNA表达水平进一步升高.结论 lncRNA TP53TG1可能参与调控牙髓炎症过程和成牙本质细胞的免疫调控作用.
Abstract
Objective To investigate the expression difference of lncRNA TP53TG1 in healthy and inflamed dental pulp and its effect on the transcription level of inflammatory factors in human dental pulp stem cells(hDPSCs)induced by lipopolysaccharide(LPS).Methods A probe set against lncRNA TP53TG1 was designed and synthesized.RNA fluorescence in situ hybridization was used to detect the expression of lncRNA TP53TG1 in healthy and inflamed dental pulp tissues.hDPSCs were isolated and cultured by tissue enzymatic digestion method,and the cultured hDPSCs were verified for multidirectional differentiation induction and identified for phenotypic characterization by flow cytometry.hDPSCs were divided into siNC group,siNC+LPS group and siTP53TG1+LPS group,which were transfected with control sequence and TP53TG1 siRNA,respectively.Then the siNC+LPS group and siTP53TG1+LPS group were treated with LPS for 24 h.The mRNA expression levels of IL-8,IL-1β3 and TNF-α in each group were detected by qRT-PCR.Results The expression of lncRNA TP53TG1 was observed in healthy dental pulp tissues,mainly distributed in odontoblasts.The expression of lncRNA TP53TG1 was increased in the inflammatory state and was distributed throughout the dental pulp.After LPS treatment,the mRNA expression levels of TNF-α,IL-1β,and IL-8 in hDPSC increased.Down-regulation of lncRNA TP53TG1 further increased the mRNA expression level of TNF-α in hDPSCs.Conclusion LncRNA TP53TG1 may be involved in the regulation of pulpitis and the immune regulation of odontoblast.
关键词
lncRNA/TP53TG1/牙髓干细胞/牙髓炎/肿瘤坏死因子-αKey words
lncRNA TP53TG1/human dental pulp stem cells/pulpitis/tumor necrosis factor-α引用本文复制引用
基金项目
国家自然科学基金(81970932)
陕西省自然科学基础研究计划重点项目(2022JZ-42)
北京市自然科学基金(7242140)
出版年
2024
国家科技期刊平台
NSTL
万方数据