氟砷联合对成骨与破骨细胞共培养体系中TRAF-6介导的NF-κB1信号通路相关蛋白表达的影响  

Effects of co-exposure of fluorine and arsenic on protein expression of TRAF-6-mediated NF-κB1 signaling pathway in osteoblasts and osteoclast co-culture systems

作　　者：杨兴 洪峰 张才良 张均涛 覃子秀 刘雅兰 金照凤 Yang Xing;Hong Feng;Zhang Cailiang;Zhang Juntao;Qin Zixiu;Liu Yalan;Jin Zhaofeng(School of Public Health,Key Laboratory for Environment Pollution Monitoring and Disease Control,Ministry of Education,Guizhou Medical University,Guiyang 550025,China;Preventive Health Care,Renhuai City People's Hospital,Zunyi 564500,China)

机构地区：[1]贵州医科大学公共卫生学院环境污染与疾病监控教育部重点实验室,贵阳550025 [2]仁怀市人民医院预防保健科,贵州遵义564500

出　　处：《中华地方病学杂志》2020年第5期318-324,共7页Chinese Journal of Endemiology

基　　金：国家自然科学基金(81472927)。

摘　　要：目的探讨氟、砷及氟砷联合染毒对小鼠成骨细胞MC3T3-E1与小鼠单核巨噬细胞RAW264.7共培养体系中肿瘤坏死因子受体相关因子6(TRAF-6)/核因子κB1(NF-κB1)信号通路中相关蛋白表达的影响.方法MC3T3-E1细胞经成骨诱导剂诱导后与RAW264.7细胞建立共培养体系,体外培养7d,采用析因设计,分别用不同剂量的氟化钠(0.0、0.1、0.4、1.6 mmol/L NaF,F)、亚砷酸钠(0.0、0.5、2.5、12.5μmol/L NaAsO2,As)以及不同剂量的氟砷联合培养基培养24h.采用蛋白免疫印迹法(Western blot)检测核因子κB受体活化因子(RANK)、TRAF-6、NF-κB1、T细胞活化因子(NFATc1)、抗酒石酸酸性磷酸酶(TRAP)的蛋白表达水平.结果氟单独作用时,与对照组(F0.0As0.0,1.00±0.00)比较,各剂量组RANK、NF-κB1、TRAP蛋白表达(1.11±0.04、1.29±0.05、1.38±0.04,1.24±0.04、1.13±0.03、1.34±0.05,1.12±0.03、1.24±0.04、1.61±0.06)增加(P均<0.05);TRAF-6蛋白表达在F01和F1.6组(1.23±0.04、1.35±0.03)增加(P均<0.05).砷单独作用时,与对照组(F0.0As0.0)比较,RANK、TRAF-6、NF-κB1蛋白表达在As0.5组增加(P均<0.05),RANK和NFATc1蛋白表达在As12.5组减少(P均<0.05).氟砷联合作用时,同一染氟剂量,RANK蛋白表达在F0.1As0.5组,TRAF-6蛋白表达在F0.1As12.5、F0.4As0.5、F04As2.5组,NF-κB1蛋白表达在F0.1As0.5、F0.4As2.5、F0.4As12.5组,NFATc1蛋白表达在F0.1As0.5、F0.4As0.5组,TRAP蛋白表达在F0.1As12.5组均高于相应的单独氟染毒组(F01、F04,P均<0.05),但低于氟、砷单独染毒之和.同一染砷剂量,RANK蛋白表达在F0.1As12.5组,TRAF-6蛋白表达在F0.1As12.5、F0.4As2.5组,NF-κB1蛋白表达在F0.1As12.5、F0.4As2.5、F0.As12.5、F1.6As2.5组,TRAP蛋白表达在F1.6As2.5、F1.6As12.5组均高于相应的单独砷染毒组(As2.5、As12.5,P均<0.05),但低于氟、砷单独染毒之和.氟对RANK、TRAF-6、NF-κB1、NFATc1、TRAP蛋白表达均有主效应作用(F=3.41、341.73、66.01、56.49、147.40,P均<0.05);砷对各蛋白指标也均有Objective To investigate the effects of combined exposure of fluorine,arsenic,and fluorine-arsenic on the signaling pathway related protein expression of tumor necrosis factor receptor-related factor 6(TRAF-6)/nuclear factorκB1(NF-κB1)in a co-culture system of mouse osteoblasts MC3T3-E1 and mouse monocyte macrophage RAW264.7.Methods MC3T3-E1 cells were co-cultured with RAW264.7 cells after induction with osteogenic inducers.The cells were cultured for 7 days in vitro,and different doses of sodium fluoride(0.0,0.1,0.4,1.6 mmol/L NaF,F),sodium arsenite(0.0,0.5,2.5,12.5μmol/L NaAsO2,As)and different doses of fluorine and arsenic were added to the culture medium and cultured for 24 h using factorial design.The expression levels of nuclear factorκB receptor activating factor(RANK),TRAF-6,NF-κB1,T cell activating factor(NFATc1),and tartrate-resistant acid phosphatase(TRAP)protein were detected by Western blotting.Results When fluorine was used alone,compared with the control group(F0.0As0.0,1.00±0.00),the expressions of RANK,NF-κB1 and TRAP proteins(1.11±0.04,1.29±0.05,1.38±0.04,1.24±0.04,1.13±0.03,1.34±0.05,1.12±0.03,1.24±0.04,1.61±0.06)were increased(P<0.05);TRAF-6 protein expressions in F0.1 and F1.6 groups(1.23±0.04,1.35±0.03)were increased(P<0.05).When arsenic was used alone,compared with the control group(F0.0As0.0),the expressions of RANK,TRAF-6,NF-κB1 proteins were increased in As0.5 group(P<0.05),the expressions of RANK and NFATc1 proteins were reduced in As12.5 group(P<0.05).When fluorine was combined with arsenic,at the same dose of fluorine,RANK protein expression in F0.1As0.5 group and TRAF-6 protein expression in F0.1As12.5,F0.4As0.5,F0.4As2.5 groups,NF-κB1 protein expression in F0.1As0.5 F0.4As2.5,F0.4As12.5 groups,NFATc1 protein expression in F0.1As0.5 and F0.4As0.5 groups,TRAP protein expression in F0.1As12.5 group were higher than the corresponding fluorine groups alone(F0.1,F0.4,P<0.05),but lower than the sum of fluorine and arsenic alone.At the same dose of arsenic,RANK protein e

关 键 词：氟 砷 联合作用 共培养 TRAF-6/NF-κB1信号通路 

分 类 号：R[医药卫生]