氟对体外培养血管内皮细胞影响及硒干预作用研究  

STUDY ON THE EFFECT OF FLUORIDE ON CULTURED ENDOTHELIAL CELLS AND THE ROLE OF SELENIUM INTERVENTION

作　　者：边建朝[1] 蔺新英[2] 杨晓霞[2] 侯小东[2] 樊婷[2] 朱秋丽[2] 

机构地区：[1]山东省地方病防治研究所,济南250014 [2]山东大学公共卫生学院营养与食品卫生教研室,济南250012

出　　处：《营养学报》2010年第5期474-479,共6页Acta Nutrimenta Sinica

基　　金：山东省人民政府、山东省国土资源厅资助(No.1212010310306)

摘　　要：目的研究不同浓度氟对体外培养血管内皮细胞的影响及硒的干预作用。方法人脐静脉血管内皮细胞（HUVEC）置于37℃,5%CO2,饱和湿度条件下培养。在培养液中加入氟化钠,氟浓度分别为0、100、400、700、1000、2000μmol/L,同时加入硒,浓度为100 nmol/L。连续培养48 h后,收集细胞培养液与细胞。应用瑞氏-吉姆萨染色观察细胞形态,吖啶橙荧光染色测定细胞凋亡,四唑氮蓝（MTT）比色法检测细胞活性;检测超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GSH-Px）活性、丙二醛（MDA）含量;检测细胞培养液诱导型一氧化氮合酶（iNOS）、内皮型一氧化氮合酶（eNOS）活性、细胞iNOSmRNA、eNOSmRNA表达;检测细胞间粘附分子-1（ICAM-1）、血管细胞粘附分子-1（VCAM-1）含量。结果在氟浓度100-700μmol/L加硒100 nmol/L,可明显减轻氟对细胞生长的抑制,恢复细胞形态的改变;使SOD、GSH-Px活性升高（P〈0.05）、MDA含量下降（P〈0.05）;降低iNOS活性和iNOSmRNA表达（P〈0.05）,升高eNOS的活性和eNOSmRNA表达（P〈0.05）;减少CAM-1和VCAM-1含量（P〈0.05,P〈0.01）。结论氟可致血管内皮细胞形态改变、功能异常。适宜剂量硒补充可通过提高抗氧化功能,调整NO代谢,抑制粘附分子异常表达等途径拮抗高氟所致的血管内皮损伤。Objective To study the influence of fluoride in different concentrations on cultured vascular endothelial cells and the effect of selenium intervention.Method Human umbilical vein endothelial cells（HUVEC） were cultivated at 37 ℃,5% CO2,saturated humidity conditions.The cell culture medium was added by sodium fluoride,at fluoride concentrations 0,100,400,1000,2000 μmol/L,while selenium was added at 100 nmol/L.Cell culture medium and cells were collected after 48h continuous incubation.Wright-Giemsa staining was applied to observe cell morphology,acridine orange fluorescence staining for cell apoptosis,and MTT colorimetry assay for cell activity.The anti-oxidation parameters as superoxide dismutase（SOD） of cytolymph,glutathione peroxidase（GSH-Px） activity,malondialdehyde（MDA） content were determined.Inducible nitric oxide synthase（iNOS） and endothelial nitric oxide synthase（eNOS） activities,cell iNOS-mRNA and eNOS-mRNA expressions and intercellular adhesion molecule（ICAM-1） and vascular adhesion molecule（VCAM-1） contents were detected.Results Adding 100 nmol/L Se to fluoride concentration of 100-700 μmol/L can reduce the inhibition of fluoride on cell growth significantly and restore the change of cell morphology.SOD and GSH-Px activities were increased（P〈0.05）,MDA content decreased（P〈0.05）,iNOS activity and iNOS-mRNA expression reduced（P〈0.05）,eNOS activity and eNOS-mRNA expression increased（P〈0.05）;and CAM-1 and VCAM-1 content reduced（P〈0.05,P〈0.01）.Conclusion Fluoride can induce vascular endothelial cell morphological changes and functional abnormalities.Selenium can improve the anti-oxidation function,adjust NO metabolism,inhibit abnormal expression of cytokines,as antagonist of vascular endothelial injury caused by high-fluoride.

关 键 词：氟 硒 人脐静脉血管内皮细胞 抗氧化酶 一氧化氮 粘附分子 

分 类 号：R599[医药卫生—内科学]