高糖对人牙龈上皮细胞氧化应激及线粒体损伤的影响  

Effects of high glucose on oxidative stress and mitochondrial damage in human gingival epithelial cells

作　　者：张钰晨 赵颖 朱春晖 裴丹丹[1,3] 苟建重 李昂[1,2] ZHANG Yuchen;ZHAO Ying;ZHU Chunhui;PEI Dandan;GOU Jianzhong;LI Ang(Key Laboratory of Shaanxi Province for Craniofacial Precision Medicine Research,Xi’an 710004,China;Department of Periodontology,Stomatology Hospital of Xi’an Jiaotong University;Department of Prosthodontics,Stomatology Hospital of Xi’an Jiaotong University)

机构地区：[1]陕西省颅颌面精准医学研究重点实验室,西安710004 [2]西安交通大学口腔医院牙周科 [3]西安交通大学口腔医院修复科

出　　处：《山西医科大学学报》2020年第9期971-977,共7页Journal of Shanxi Medical University

基　　金：国家自然科学基金资助项目(81901019,81870798)。

摘　　要：目的探索高糖微环境对人牙龈上皮细胞(human gingival epithelial cells,HGECs)氧化应激及线粒体损伤的影响。方法将培养的HGECs随机分成正常对照组、等渗组、高糖组,体外培养48 h。2′,7′-二氯二氢荧光素双乙酸酯探针检测细胞内活性氧含量。JC-1荧光染色检测HGECs线粒体膜电位变化。qRT-PCR检测细胞抗氧化酶SOD1、GCLC、CAT的mRNA水平。免疫荧光法检测细胞内pNrf2、COXⅣ的表达。线粒体绿色荧光探针检测细胞线粒体数量。结果与正常对照组及等渗组相比,高糖组HGECs的活性氧水平显著上升(P<0.05),pNrf2表达水平及抗氧化酶基因表达显著降低(P<0.05),线粒体膜电位下降,线粒体生物合成及线粒体数量减少。结论高糖微环境可诱导HGECs损伤,其机制可能与细胞发生氧化应激和线粒体损伤有关。Objective To explore the effects of high glucose on oxidative stress and mitochondrial damage in human gingival epithelial cells(HGECs).Methods HGECs were randomly divided into three groups:normal control group,isotonic group,high glucose group,and then HGECs were incubated for 48 h,respectively.The intracellular reactive oxygen species(ROS)content was detected using the fluorescent 2′,7′-dichlorodihydrofluorescein diacetate probe.The change of mitochondrial membrane potential was investigat-ed by JC-1 fluorescent staining.The mRNA levels of antioxidant enzymes SOD1,GCLC,CAT were quantified by qRT-PCR.The expression of phospho-Nrf2 and COXⅣwas detected by immunofluorescence technique.Mitotracker green was used to measure the number of mitochondria.Results Compared with normal control group and isotonic group,the content of ROS in HGECs was significantly increased in high glucose group(P<0.05),and the expression levels of phospho-Nrf2 and its downstream antioxidant enzyme genes(SOD1,GCLC,CAT)in HGECs were significantly decreased after high glucose stimulation for 48 h(P<0.05).Simultaneously,the mitochondrial membrane potential was decreased and the biogenesis and number of mitochondria were reduced in high glucose group.Conclusion High glucose microenvironment could induce the damage of HGECs,which may be related to oxidative stress and mitochondrial damage.

关 键 词：高糖 人牙龈上皮细胞 氧化应激 线粒体损伤 

分 类 号：R363[医药卫生—病理学]