阿魏酸对大气细颗粒物PM2.5诱导的小鼠主动脉炎症的干预作用
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摘要
目的探讨大气细颗粒物PM2.5对小鼠主动脉Toll样受体(Toll-like receptors,TLRs)信号通路的影响及阿魏酸的干预作用。方法采用气管滴注方法以10,20 mg·kg~(-1)的PM2.5处理小鼠,并用40,80 mg·kg~(-1)的阿魏酸对部分20 mg·kg~(-1)PM2.5处理后的小鼠进行治疗。2周后处死小鼠,血液细胞分析仪检测小鼠血液中炎症细胞水平,ELISA检测血清中IL-1β和IL-6的含量,并用荧光定量PCR检测主动脉中IL-1β和IL-6的m RNA水平,Western blot检测各组小鼠主动脉组织中TLR2、TLR4、My D88、NF-κB p65蛋白的表达,免疫组化技术观察TLR2和TLR4在小鼠主动脉中的水平。结果PM2.5能提高血液中NEUT、EOS的数量及其在总细胞中的比率,增加血液和主动脉组织中炎症因子IL-1β、IL-6的含量,上调主动脉组织中的TLRs通路相关分子TLR2、TLR4、My D88、NF-κB p65的表达;而用阿魏酸干预后,血液中白细胞数量、EOS数量、NEUT与EOS的比率明显降低,IL-1β、IL-6水平下降。同时主动脉组织中的TLRs通路相关分子的表达下调。结论PM2.5可能通过TLRs通路诱导小鼠主动脉炎症的产生,而阿魏酸可能通过抑制TLRs通路相关因子的表达而发挥抑制PM2.5诱导的小鼠主动脉炎症的作用。
OBJECTIVE To investigate the effect of Toll-like receptors(TLRs)signaling pathway on PM2.5-induced inflammation in mice aorta and the intervention of ferulic acid.METHODS Mice were treated with 10,20 mg·kg~(-1) PM2.5respectively by onsurgical intratracheal instillation.Additionally,mice were concurrently treated with 40,80 mg·kg~(-1) ferulic acid at the high dose group(20 mg·kg~(-1) PM2.5)for 2 weeks.The categories of inflammatory cells and inflammatory factors in serum were detected by hemathology analyzer.Isolated aortic tissue was isolated and the m RNA expression of IL-1βand IL-6 were determined by RT-PCR.The protein level of TLR2,TLR4,My D88 and NF-κB p65 were detected by Western blot.The expression of TLR2 and TLR4 in mice aorta was also evaluated by immunohistochemistry.RESULTS The content and the ratio of NEUT and EOS were increased by PM2.5.The m RNA levels of IL-1?and IL-6 were overexpressed in both blood and aortic tissue.Meanwhile,TLRs signaling pathway-related protein such as TLR2,TLR4,My D88,and NF-?B p65 were also increased in aortic tissue.After the intervention of ferulic acid,the numbers of NEUT,EOS,and the ratio of NEUT and EOS,as well as the level of inflammation factors including IL-6 and IL-1?were reduced.Moreover,the inhibition TLRs pathway in aortic tissue were also significantly reduced.CONCLUSION PM2.5 can induce the production of aortic inflammation in mice through TLRs signaling pathway.Moreover,ferulic acid can protect mice from PM2.5-induced aortic inflammation by inhibition TLRs pathway.
OBJECTIVE To investigate the effect of Toll-like receptors(TLRs)signaling pathway on PM2.5-induced inflammation in mice aorta and the intervention of ferulic acid.METHODS Mice were treated with 10,20 mg·kg~(-1) PM2.5respectively by onsurgical intratracheal instillation.Additionally,mice were concurrently treated with 40,80 mg·kg~(-1) ferulic acid at the high dose group(20 mg·kg~(-1) PM2.5)for 2 weeks.The categories of inflammatory cells and inflammatory factors in serum were detected by hemathology analyzer.Isolated aortic tissue was isolated and the m RNA expression of IL-1βand IL-6 were determined by RT-PCR.The protein level of TLR2,TLR4,My D88 and NF-κB p65 were detected by Western blot.The expression of TLR2 and TLR4 in mice aorta was also evaluated by immunohistochemistry.RESULTS The content and the ratio of NEUT and EOS were increased by PM2.5.The m RNA levels of IL-1?and IL-6 were overexpressed in both blood and aortic tissue.Meanwhile,TLRs signaling pathway-related protein such as TLR2,TLR4,My D88,and NF-?B p65 were also increased in aortic tissue.After the intervention of ferulic acid,the numbers of NEUT,EOS,and the ratio of NEUT and EOS,as well as the level of inflammation factors including IL-6 and IL-1?were reduced.Moreover,the inhibition TLRs pathway in aortic tissue were also significantly reduced.CONCLUSION PM2.5 can induce the production of aortic inflammation in mice through TLRs signaling pathway.Moreover,ferulic acid can protect mice from PM2.5-induced aortic inflammation by inhibition TLRs pathway.
引文
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[26]NOREEN M,ARSHAD M.Association of TLR1,TLR2,TLR4,TLR6,and TIRAP polymorphisms with disease susceptibility[J].Immunol Res,2015,62(2):234-252.
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[28]CHUKKAPALLI S S,VELSKO I M,RIVERA-KWEH M F,et al.Global TLR2 and 4 deficiency in mice impacts bone resorption,inflammatory markers and atherosclerosis to polymicrobial infection[J].Mol Oral Microbiol,2017,32(3):211-225.
[29]EDFELDT K,SWEDENBORG J,HANSSON G K,et al.Expression of toll-like receptors in human atherosclerotic lesions:a possible pathway for plaque activation[J].Circulation,2002,105(10):1158-1161.
[30]OGIWARA T,SATOH K,KADOMA Y,et al.Radical scavenging activity and cytotoxicity of ferulic acid[J].Anticancer Res,2002,22(5):2711-2717.
[31]LIU Y M,SHEN J D,XU L P,et al.Ferulic acid inhibits neuro-inflammation in mice exposed to chronic unpredictable mild stress[J].Int Immunopharmacol,2017(45):128-134.
[32]YUAN X,WANG Y,DU D,et al.The effects of the combination of sodium ferulate and oxymatrine on lipopolysaccharide-induced acute lung injury in mice[J].Inflammation,2012,35(3):1161-1168.
[33]LEE C C,WANG C C,HUANG H M,et al.Ferulic acid induces Th1 responses by modulating the function of dendritic cells and ameliorates Th2-mediated allergic airway inflammation in mice[J].Evid Based Complement Alternat Med,2015(2015):678487.
[34]NIU C W,SHENG Y C,ZHU E Y,et al.Ferulic acid prevents liver injury induced by Diosbulbin B and its mechanism[J].Bio Sci Trends,2016,10(5):386-391.
[2]KULICK E R,WELLENIUS G A,KAUFMAN J D,et al.Long-term exposure to ambient air pollution and subclinical cerebrovascular disease in NOMAS(the northern manhattan study)[J].Stroke,2017,48(7):1966-1968.
[3]GOULOPOULOU S,MCCARTHY C G,WEBB R C.Toll-like receptors in the vascular system:sensing the dangers within[J].Pharmacol Rev,2016,68(1):142-167.
[4]胡益勇,徐晓玉.阿魏酸的化学和药理研究进展[J].中成药,2006,28(2):253-255.
[5]OJHA S,JAVED H,AZIMULLAH S,et al.Neuroprotective potential of ferulic acid in the rotenone model of Parkinson's disease[J].Drug Des Devel Ther,2015(9):5499-5510.
[6]QIAN X,ZHANG L,HUANG W F,et al.Determination of chlorogenic acid and ferulic acid in Gouqigen by HPLC[J].Pharm Today(今日药学),2016,26(6):415-417.
[7]NAVARRETE S,ALARCóN M,PALOMO I.Aqueous extract of tomato(Solanum lycopersicum L.)and ferulic acid reduce the expression of TNF-αand IL-1βin LPS-activated macrophages[J].Molecules,2015,20(8):15319-15329.
[8]ZHOU Y L,LAO W Y,RUAN Y S,et al.Protective effect of ferulic acid on lung injury induced by PM2.5 in rats[J].Food Sci(食品科学),2017,38(1):244-251.
[9]BAI R,ZHANG L,LIU Y,et al.Pulmonary responses to printer toner particles in mice after intratracheal instillation[J].Toxicol Lett,2010,199(3):288-300.
[10]FRANKLIN B A,BROOK R,ARDEN POPE C.Air pollution and cardiovascular disease[J].Curr Probl Cardiol,2015(40):207-238.
[11]LIU S T,LIAO C Y,KUO C Y,et al.The effects of PM2.5from asian dust storms on emergency room visits for cardiovascular and respiratory diseases[J].Int J Environ Res Public Health,2017,14(4):E428.
[12]APTE J S,MARSHALL J D,COHEN A J,et al.Addressing global mortality from ambient PM2.5[J].Environ Sci Technol,2015,49(13):8057-8066.
[13]GHIO A,DEVLIN R.Inflammatory lung injury after bronchial instillation of air pollution particles[J].Am J Respir Crit Care Med,2001,164(4):704-708.
[14]GHIO A,HALL A,BASSETT M,et al.Exposure to concentrated ambient air particles alters hematologic indices in humans[J].Inhal Toxicol,2003,15(14):1465-1478.
[15]GRAFF D,CASCIO W,RAPPOLD A,et al.Exposure to concentrated coarse air pollution particles causes mild cardiopulmonary effects in healthy young adults[J].Environ Health Perspect,2009,117(7):1089-1094.
[16]SANGANI R G,SOUKUP J M,GHIO A J.Metals in air pollution particles decrease whole-blood coagulation time[J].Inhal Toxicol,2010,22(8):621-626.
[17]POPE C A,BHATNAGAR A,MCCRACKEN J P,et al.Exposure to fine particulate air pollution is associated with endothelial injury and systemic inflammation[J].Circ Res,2016,119(11):1204-1214.
[18]GOUILL E L,JIMENEZ M,BINNERT C,et al.Endothelial nitric oxide synthase(e NOS)knockout mice have defective mitochondrial beta-oxidation[J].Diabetes,2007,56(11):2690-2696.
[19]李磊,戴敏.动脉粥样硬化血管内皮分泌功能失调与平滑肌细胞增殖[J].中国药理学通报,2010,26(2):155-158.
[20]RUNDELL K W,HOFFMAN J R,CAVISTON R,et al.Inhalation of ultrafine and fine particulate matter disrupts systemic vascular function[J].Inhal Toxicl,2007,19(2):133-l40.
[21]POPE C A,BHATNAGAR A,MCCRACKEN J P,et al.Exposure to fine particulate air pollution is associated with endothelial injury and systemic inflammation[J].Circ Res,2016,119(11):1204-1214.
[22]MONTIEL-DáVALOS A,IBARRA-SáNCHEZ M J,VENTURA-GALLEGOS J L,et al.Oxidative stress and apoptosis are induced in human endothelial cells exposed to urban particulate matter[J].Toxicol In Vitro,2010,24(1):135-141.
[23]LI R,NING Z,MAJUMDAR R,CUI J,et al.Ultrafine particles from diesel vehicle emissions at different driving cycles induce differential vascular pro-inflammatory responses:Implication of chemical components and NF-κB signaling[J].Part Fibre Toxicol,2010,7(1):6-18.
[24]COLE J E,GEORGIOU E,MONACO C.The expression and functions of toll-like receptors in atherosclerosis[J].Mediators Inflamm,2010(2010):393946.Doi:10.1155/2010/393946.
[25]GHOSH S,DASS J F.Study of pathway cross-talk interactions with NF-κB leading to its activation via ubiquitination or phosphorylation:A brief review[J].Gene,2016,584(1):97-109.
[26]NOREEN M,ARSHAD M.Association of TLR1,TLR2,TLR4,TLR6,and TIRAP polymorphisms with disease susceptibility[J].Immunol Res,2015,62(2):234-252.
[27]ROSHAN M H,TAMBO A,PACE N P.The role of TLR2,TLR4,and TLR9 in the pathogenesis of atherosclerosis[J].Int J Inflam,2016,2016:1532832.Doi:10.1155/2016/1532832
[28]CHUKKAPALLI S S,VELSKO I M,RIVERA-KWEH M F,et al.Global TLR2 and 4 deficiency in mice impacts bone resorption,inflammatory markers and atherosclerosis to polymicrobial infection[J].Mol Oral Microbiol,2017,32(3):211-225.
[29]EDFELDT K,SWEDENBORG J,HANSSON G K,et al.Expression of toll-like receptors in human atherosclerotic lesions:a possible pathway for plaque activation[J].Circulation,2002,105(10):1158-1161.
[30]OGIWARA T,SATOH K,KADOMA Y,et al.Radical scavenging activity and cytotoxicity of ferulic acid[J].Anticancer Res,2002,22(5):2711-2717.
[31]LIU Y M,SHEN J D,XU L P,et al.Ferulic acid inhibits neuro-inflammation in mice exposed to chronic unpredictable mild stress[J].Int Immunopharmacol,2017(45):128-134.
[32]YUAN X,WANG Y,DU D,et al.The effects of the combination of sodium ferulate and oxymatrine on lipopolysaccharide-induced acute lung injury in mice[J].Inflammation,2012,35(3):1161-1168.
[33]LEE C C,WANG C C,HUANG H M,et al.Ferulic acid induces Th1 responses by modulating the function of dendritic cells and ameliorates Th2-mediated allergic airway inflammation in mice[J].Evid Based Complement Alternat Med,2015(2015):678487.
[34]NIU C W,SHENG Y C,ZHU E Y,et al.Ferulic acid prevents liver injury induced by Diosbulbin B and its mechanism[J].Bio Sci Trends,2016,10(5):386-391.