摘要
目的研究牙龈卟啉单胞菌(Pg)脂多糖(LPS)对牙周膜间充质干细胞(PDLSCs)组蛋白甲基化转移酶PRDM9的作用及调控机制。方法选择不同浓度的Pg. LPS处理PDLSCs;利用实时定量RT-PCR的方法检测多种组蛋白甲基化转移酶在mRNA水平的表达变化;此外在Pg. LPS处理细胞的同时加入0.5μg/ml TLR4中和抗体,比较阻断及未阻断情况下筛选出的差异基因在mRNA水平的表达变化。结果实时定量RT-PCR结果显示PDLSCs在10μg/ml Pg. LPS刺激24 h和48 h后,组蛋白甲基化转移酶PRDM9基因表达量显著升高,具有时间相关性。选用1、5和10μg/ml的Pg. LPS处理PDLSCs,48h后检测到PRDM9表达量升高与Pg. LPS呈剂量相关性;与对照组相比阻断TLR4信号通路使得PRDM9的表达量明显下调。结论 Pg. LPS能够激活TLR4信号通路进而促进组蛋白甲基化转移酶PRDM9基因的表达。
Objective To investigate the effect of Porphyromonasgingivalis(Pg)lipopolysaccharide(LPS)on histone methylation transferase in periodontal ligament mesenchymal stem cells(PDLSCs)and its regulatory mechanism.Methods Different dose of Pg.LPS was used to stimulate the PDLSCs.Real time RT-PCR was used to detect the expression of histone methyltransferase at the mRNA level.In addition,0.5μg/ml TLR4 neutralizing antibody was added to Pg.LPStreated PDLSCs at the same time to compare the expression changes of differentially screened genes at the mRNA level.Results Real time RT-PCR results showed that the expression of PRDM9 was up-regulated at 24 and 48h after Pg.LPS treatment in a time dependent manner.And 1,5 and 10μg/ml Pg.LPS could increase the expression level of PRDM9 in a dose dependent manner at 48h.Compared with the control group,the expression of PRDM9 decreased significantly after the application of TLR4 neutralizing antibody.Conclusion Pg.LPS can activate TLR4 signaling pathway and promote the expression of histone methyltransferase PRDM9 gene in PDLSCs.
作者
张建鹏
张琛
刘惠娜
侯本祥
ZHANG Jian-peng;ZHANG Chen;LIU Hui-na;HOU Ben-xiang(Department of Endodontics,Capital Medical University School of Stomatology,Beijing 100050,China)
出处
《北京口腔医学》
CAS
2020年第4期203-207,共5页
Beijing Journal of Stomatology
基金
国家自然科学基金(81170952)
北京口腔医院学科建设基金(18-09-07)。
