王佳;孙霞;董四君;

• 1:中国科学院城市环境研究所环境与健康重点实验室

摘要(Abstract)：

为探讨邻苯二甲酸二乙基己酯((DEHP)的细胞免疫毒性作用与机制,采用RT-PCR和ELISA方法,考察了0.05～1μmol·L-1浓度范围内的DEHP对THP-1细胞白细胞介素-1β(IL-1β)及基质金属蛋白酶-8(MMP-8)基因和蛋白表达的影响;采用免疫印迹WesternBlot方法检测DE-HP对ERK1/2磷酸化水平的影响;以2,'7-'二氯荧光素二乙酸酯(DCFH-DA)为荧光探针检测1～50μmol·L-1DEHP对细胞内活性氧(ROS)产生的影响。结果显示,0.05和0.2μmol·L-1DEHP在6h内显著诱导IL-1β和MMP-8基因表达(P<0.05或0.01);0.05～1μmol·L-1DEHP刺激细胞48h,可诱导IL-1β蛋白表达,并表现出明显的剂量-效应关系,线性拟合的确定系数为0.937;0.05μmol·L-1DEHP刺激细胞6或12h,显著诱导MMP-8蛋白表达(P<0.05);0.2μmol·L-1DEHP在15～30min内快速诱导ERK1/2磷酸化;1～50μmol·L-1DEHP浓度依赖性刺激细胞中ROS的产生;ERK/MAPK抑制剂PD98059显著抑制DEHP诱导的MMP-8分泌,但对IL-1β分泌未表现出抑制作用。研究表明,DEHP可能是经ERK/MAPK信号路径诱导MMP-8基因和蛋白的表达,经其他路径诱导IL-1β基因和蛋白的表达,诱导细胞内ROS的产生,从而激发炎症反应,进而损害免疫系统功能引发哮喘等炎症性疾病。此研究结果可为DEHP的暴露风险评估提供参考。

关键词(KeyWords)： DEHP;炎症;免疫;过敏;哮喘;ERK/MAPK;IL-1β;MMP-8

Abstract:

Keywords:

基金项目(Foundation): 中国科学院百人计划项目(KZCX2-YW-BR-18);; 厦门市科技计划项目(3502Z20112019)

作者(Author): 王佳;孙霞;董四君;

Email:

参考文献(References)：

• [1]Heudorf U,Mersch-Sundermann V,Angerer J.Phthalates:Toxicology and exposure[J].International Journal of Hygiene and Environmental Health,2007,210(5):623-634
• [2]Rudel R A,Gray J M,Engel C L,et al.Food packa-ging and bisphenol A and bis(2-ethyhexyl)phthalate exposure:Findings from adietary intervention[J].Envi-ronmental Health Perspectives,2011,119(7):914-920
• [3]Shaz B H,Grima K,Hillyer C D.2-(Diethylhexyl)phthalate in blood bags:Is this a public health issue?[J].Transfusion,2011,51(11):2510-2517
• [4]Chen S Q,Chen J N,Cai X H,et al.Perinatal expo-sure to di-(2-ethylhexyl)phthalate leads to restricted growth and delayed lung maturation in newborn rats[J].Journal of Perinatal Medicine,2010,38(5):515-521
• [5]唐文红.丙泊酚对脂多糖刺激人单核细胞分泌细胞因子的影响[D].上海:第二军医大学,2008:6Tang W H.Effect of propofol on the secretion of cyto-kines from human monocytes stimulated with LPS[D].Shanghai:The Second Military Medical University,2008:6(in Chinese)
• [6]Nishioka J,Iwahara C,Kawasaki M,et al.Di-(2-eth-ylhexyl)phthalate induces production of inflammatory molecules in human macrophages[J].Inflammation Research,2012,61(1):69-78
• [7]Garcia-Rio F,Miravitlles M,Soriano J B,et al.Sys-temic inflammation in chronic obstructive pulmonary dis-ease:A population-based study[J].Respiratory Re-search,2010,11(1):63-78
• [8]Mattoli S,Marini M,Fasoli A.Expression of the potent inflammatory cytokines,GM-CSF,IL6,and IL8,in bronchial epithelial cells of asthmatic patients[J].Chest,1992,101(3s):27S-29S
• [9]Prikk K,Maisi P,Piril E,et al.Airway obstruction correlates with collagenase-2(MMP-8)expression and activation in bronchial asthma[J].Laboratory Investi-gation,2002,82(11):1535-1545
• [10]Liu W,Liang Q,Balzar S,et al.Cell-specific activa-tion profile of extracellular signal-regulated kinase1/2,Jun N-terminal kinase,and p38mitogen-activated pro-tein kinases in asthmatic airways[J].Journal of Allergy and Clinical Immunology,2008,121(4):893-902.e2
• [11]Pak V M,McCauley L A.Risks of phthalate exposure among the general population:Implications for occupa-tional health nurses[J].Official Journal of the Ameri-can Association of Occupational Health Nurses,2007,55(1):12-17
• [12]Cargnello M,Roux P P.Activation and function of the MAPKs and their substrates,the MAPK-activated pro-tein kinases[J].Microbiology and Molecular Biology Reviews,2011,75(1):50-83
• [13]Halden R U.Plastics and health risks[J].Annual Re-view of Public Health,2010,31(1):179-194
• [14]Lee J,Oh P S,Lim K T.Allergy-related cytokines(IL-4and TNF-α)are induced by di(2-ethylhexyl)phthalate and attenuated by plant-originated glycoprotein(75kDa)in HMC-1cells[J].Environmental Toxicol-ogy,2011,26(4):364-372
• [15]Van Lint P,Libert C.Matrix metalloproteinase-8:Cleavage can be decisive[J].Cytokine&Growth Fac-tor Reviews,2006,17(4):217-223
• [16]Obase Y,Rytila P,Metso T,et al.Effects of inhaled corticosteroids on metalloproteinase-8and tissue inhibi-tor of metalloproteinase-1in the airways of asthmaticchildren[J].International Archives of Allergy and Im-munology,2010,151(3):247-254
• [17]Zhang H.Anti-IL-1beta therapies[J].Recent Patents on DNA&Gene Sequences,2011,5(2):126-135
• [18]Oh P S,Lim K,Lim K T.Phytoglycoprotein(75kDa)inhibits expression of interleukin-1beta stimulated by DEHP in human mast cells[J].Cell Biochemistry and Function,2010,28(5):352-359
• [19]蒋国新.哮喘患者血清NO、IL-1β水平变化及其与肺功能的相关性研究[J].交通医学,2009,23(5):485-487Jiang G X.Correlation study between serum nitrogen mon-oxidum,interleukin-1βlevels and pulmonary function in asthma[J].Medical Journal of Communications,2009,23(5):485-487(in Chinese)
• [20]Wang J,Dong S.ICAM-1and IL-8are expressed by DEHP and suppressed by curcumin through ERK and p38MAPK in human umbilical vein endothelial cells[J].Inflammation,2012,35(3):859-870
• [21]Shan L,Redhu N S,Saleh A,et al.Thymic stromal lymphopoietin receptor-mediated IL-6and CC/CXC che-mokines expression in human airway smooth muscle cells:Role of MAPKs(ERK1/2,p38,and JNK)and STAT3pathways[J].The Journal of Immunology,2010,184(12):7134-7143
• [22]Goplen N,Karim Z,Guo L,et al.ERK1is important for Th2differentiation and development of experimental asthma[J].The Journal of the Federation of American Societies of Experimental Biology,2012,26(5):1934-1945
• [23]Quercioli A,Mach F,Bertolotto M,et al.Receptor ac-tivator of NF-κB ligand(RANKL)increases the release of neutrophil products associated with coronary vulnera-bility[J].Thrombosis and Haemostasis,2012,107(1):124-139
• [24]van der Vliet A.Nox enzymes in allergic airway inflam-mation[J].Biochimica et Biophysica Acta(BBA)-General Subjects,2011,1810(11):1035-1044