炎症微环境下牙周膜干细胞胞内ROS的表达  

Expression of reactive oxygen species in human periodontal ligament stem cells under an inflammatory microenvironment

作　　者：王文娟[1] 严妍[2] 张凤秋[3] WANG Wen-juan;YAN Yan;ZHANG Feng-qiu(Department of General Denristry,Beijing Stomatological Hospital,Capital Medical University,Beijing 100050,China)

机构地区：[1]首都医科大学口腔医学院王府井部口腔综合科,北京100050 [2]首都医科大学口腔医学院王府井部牙体牙髓科,北京100050 [3]首都医科大学口腔医学院王府井部牙周科,北京100050

出　　处：《北京口腔医学》2022年第3期171-175,共5页Beijing Journal of Stomatology

基　　金：国家自然科学基金(81500852)。

摘　　要：目的探讨脂多糖(LPS)诱导的炎症微环境下,牙周膜干细胞胞内ROS表达水平与牙周膜干细胞成骨向分化能力的关系。方法分别用DMEM培养基(阴性对照组)、成骨诱导液(阳性对照组)、成骨诱导液+10μg/ml的LPS(实验组)、成骨诱导液+10μg/ml LPS+20μmol/l白皮杉醇(实验组)处理牙周膜干细胞0、7、14、21 d后收集细胞,分别用实时定量PCR的方法检测Runx2,ALP以及OCN的表达,用流式细胞仪检测胞内ROS水平的变化,收集连续培养21 d的牙周膜干细胞进行茜素红染色,检测牙周膜干细胞成骨分化的结果。结果LPS组Runx2,ALP和OCN的表达远低于成骨诱导组,其差异具有统计学差异(P<0.05)。白皮杉醇组Runx2,ALP和OCN远高于LPS组,其差异具有统计学差异(P<0.05),但与成骨诱导组相比无统计学差异(P>0.05)。21 d茜素红染色的结果与基因水平结果一致。在同一时间点内,牙周膜干细胞胞内ROS的表达水平在成骨诱导+LPS组水平最高,成骨诱导组最低,成骨诱导+LPS+白皮杉醇胞内ROS水平介于成骨诱导组和成骨诱导液+LPS组之间,但数值更接近于成骨诱导组。结论白皮杉醇通过抑制牙周膜干细胞胞内ROS水平的表达,缓解LPS对成骨相关指标的抑制作用。Objective To investigate the relationship between reactive oxygen species(ROS)in human periodontal ligament stem cells(hPDLSCs)and the osteogenesis gene expression under an inflammatory microenvironment induced by lipopolysaccharide(LPS).Methods The hPDLSCs were cultured with Dulbecco's Modified Eagle's medium(DMEM),osteogenic induction medium,osteogenic induction medium+10μg/ml LPS,and osteogenic induction medium+10μg/ml LPS+20μmol/l piceatannol respectively,and collected on 0,7,14,21 days later.Then,the level of Runx2,ALP and OCN genes was detected using real-time quantitative PCR,and the level of ROS inside hPDLSCs was measured by flow cytometry.hPDLSCs were cultured for 21 days and stained with Alizarin Red S.Results The group cultured with osteogenic induction medium+10μg/ml LPS showed significant lower expression level of Runx2,ALP and OCN than the osteogenic induction medium group(P<0.05).The results of 21 days with Alizarin Red S staining were consistent with gene level results.Meanwhile,the osteogenic induction medium+10μg/ml LPS+20μmol/l piceatannol group showed significant higher expression level of these genes than osteogenic induction medium+10μg/ml LPS group(P<0.05).However,it did not show a statistical difference compared with the osteogenic induction medium group(P>0.05).After cultured for certain periods,the osteogenic induction medium+10μg/ml LPS group demonstrated the highest level of ROS inside hPDLSCs,while the osteogenic induction medium group showed the lowest.Furthermore,osteogenic induction medium+10μg/ml LPS+20μmol/l piceatannol group showed a medium ROS level.Conclusions Piceatannol might mitigate the depression on the osteogenesis gene expression caused by LPS by lowering the level of ROS inside hPDLSCs.

关 键 词：脂多糖 牙周膜干细胞 成骨向分化 活性氧 白皮杉醇 

分 类 号：R782[医药卫生—口腔医学]