炎症微环境对牙周膜干细胞氧化应激和线粒体生成的影响

Effects of inflammatory microenvironment on the oxidative stress and mitochondriogenesis of human periodontal ligament stem cells

目的:探讨炎症微环境对牙周膜干细胞(PDLSCs)氧化应激和线粒体生成的影响。方法:体外分离因正畸治疗需要而拔除的健康牙齿及慢性牙周炎患牙的PDLSCs,分组培养健康PDLSCs(H-PDLSCs)、炎症PDLSCs(P-PDLSCs)和肿瘤坏死因子α作用下的PDLSCs(T-PDLSCs)。培养7 d后,采用荧光探针和流式细胞技术检测线粒体及全细胞的活性氧簇(ROS)水平,采用实时定量PCR和Western blot技术分别检测线粒体生成及抗氧化相关基因的mRNA和蛋白表达水平。结果:与H-PDLSCs相比,P-PDLSCs和T-PDLSCs组细胞线粒体和全细胞ROS水平显著增加,线粒体生成相关基因ERRα、PGC-1α、TIMM13、MFN2和抗氧化基因PRDX3与SOD2的mRNA表达水平均显著降低,ERRα、PGC-1α、PGC-1β和SOD2蛋白表达水平均显著降低。结论:炎症微环境显著提高PDLSCs的氧化应激水平,抑制PDLSCs的线粒体生成和抗氧化反应。

Objective:To investigate the effects of inflammatory microenvironment on the oxidative stress and mitochondriogenesis of human periodontal ligament stem cells(PDLSCs).Methods:PDLSCs were obtained from healthy individuals and patients with periodontitis(H-PDLSCs,P-PDLSCs)and another group of H-PDLSCs were cultured under the treatment of tumor necrosis factor-α(10 ng/mL)(T-PDLSCs).After cultured for 7 days,the mitochondrial and the whole-cell reactive oxygen species(ROS)were detected by fluorescence probe and flow cytometry,the mRNA and protein expression of genes related to mitochondriogenesis and oxidation resistance were determined by real-time PCR and Western Blot respectively.Results:Compared to H-PDLSCs,both P-PDLSCs and T-PDLSCs showed significantly increased mitochondrial and whole-cell ROS level,decreased mRNA expression of mitochondriogenesis-related genes ERRα,PGC-1α,TIMM13,MFN2 and antioxidative genes PRDX3 and SOD2,and decreased protein level of ERRα,PGC-1α,PGC-1βand SOD2.Conclusion:Inflammatory microenvironment significantly improved oxidative stress and inhibited the mitochondriogenesis and antioxidant response of PDLSCs.