交通相关PM_(2.5)不同组分对哮喘大鼠肺部炎症及外周血中Treg细胞比例的影响
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摘要
目的研究交通相关PM_(2.5)不同组分对哮喘大鼠肺部炎症、Treg细胞及其相关细胞因子分泌水平的影响,以探讨PM_(2.5)不同组分诱导哮喘加重的机制。方法通过卵清蛋白腹腔注射致敏、雾化吸入激发构建大鼠哮喘模型,激发后分别予以3 mg/kg PM_(2.5)和1.8、7.2 mg/kg PM_(2.5)水溶组分及0.6、2.4 mg/kg PM_(2.5)有机组分气管滴注染毒,每3 d一次,共5次。观察肺组织病理改变,以流式细胞术检测外周血Treg细胞的比例。收集支气管肺泡灌洗液(BALF),以ELISA法检测BALF中IL-10和TGF-β1的含量。结果与生理盐水对照组、哮喘组、哮喘+PM_(2.5)组、哮喘+1.8 mg/kg PM_(2.5)水溶组分染毒组相比,哮喘+7.2 mg/kg PM_(2.5)水溶组分染毒组外周血中Treg细胞比例明显降低(P<0.05);哮喘+0.6 mg/kg PM_(2.5)有机组分染毒组外周血中Treg细胞比例明显低于哮喘+DMSO溶剂对照组和哮喘+PM_(2.5)组(P<0.05)。与生理盐水对照组、哮喘组、哮喘+PM_(2.5)组相比,哮喘+7.2 mg/kg PM_(2.5)水溶组分染毒组BALF中IL-10的含量明显升高(P<0.05);哮喘+0.6 mg/kg PM_(2.5)有机组分染毒组BALF中IL-10的含量低于哮喘+PM_(2.5)组(P<0.05)。哮喘+PM_(2.5)组BALF中TGF-β1表达明显高于哮喘组和哮喘+1.8 mg/kg PM_(2.5)水溶组分染毒组(P<0.05);与哮喘+DMSO溶剂对照组相比,哮喘+PM_(2.5)组、哮喘+0.6 mg/kg PM_(2.5)有机组分染毒组、哮喘+2.4 mg/kg PM_(2.5)有机组分染毒组BALF中TGF-β1表达明显升高(P<0.05)。结论交通相关PM_(2.5)水溶组分和有机组分可抑制哮喘大鼠Treg细胞的增殖分化,引起其分泌的相关细胞因子的改变,进而加重哮喘肺部炎症反应。
Objective To understand the effects of different components of traffic-related PM_(2.5) on the lung inflammation and the ratio of Treg cells and the levels of corresponding cytokines in rats with asthma,and explore the mechanism of asthma exacerbation induced by different components of PM_(2.5). Methods The rats with asthma were sensitized by intraperitoneal injections of ovalbumin and challenged by aerosol inhalation of ovalbumin,then were administrated intratracheally with 3 mg/kg PM_(2.5),and 1.8,7.2 mg/kg PM_(2.5) water soluble components and 0.6,2.4 mg/kg PM_(2.5) organic components once every three days for five times.The pathological changes were observed by HE staining of lung tissue pathological sections. The ratio of Treg cells in peripheral blood were detected by flow cytometry. The levels of IL-10 and TGF-β in bronchoalveolar lavage fluid(BALF) were determined by ELISA. Results Compared with saline control group,asthma group,asthma+PM_(2.5) group and asthma+1.8 mg/kg PM_(2.5) water soluble extracts group,the ratio of Treg cells in peripheral blood was significantly decreased in asthma+7.2 mg/kg PM_(2.5) water soluble extracts group(P<0.05);The ratio of Treg cells in peripheral blood of asthma + 1.8 mg/kg PM_(2.5) organic extracts group was significantly decreased compared with that in asthma + DMSO group and asthma + PM_(2.5) group(P<0.05). Compared with saline control group,asthma group and asthma + PM_(2.5) group,the level of IL-10 in BALF in asthma+7.2 mg/kg PM_(2.5) water soluble extracts group was significantly increased(P<0.05);The level of IL-10 in BALF in asthma+0.6 mg/kg PM_(2.5) organic extracts group was decreased compared with that in asthma + PM_(2.5) group(P<0.05). The level of TGF-β1 in BALF in asthma +PM_(2.5) group was significantly higher than those in asthma group and asthma+1.8 mg/kg PM_(2.5) water soluble extracts group(P<0.05);The levels of TGF-β1 in BALF in asthma + PM_(2.5) group,asthma + 0.6 mg/kg PM_(2.5) organic extracts group and asthma+2.4 mg/kg PM_(2.5) organic extracts group was increased compared with that in asthma + DMSO group(P<0.05). Conclusion The water soluble extracts and organic extracts of traffic-related PM_(2.5) can suppress the proliferation and differentiation of Treg,and cause the changes of corresponding cytokines to aggravate asthma.
Objective To understand the effects of different components of traffic-related PM_(2.5) on the lung inflammation and the ratio of Treg cells and the levels of corresponding cytokines in rats with asthma,and explore the mechanism of asthma exacerbation induced by different components of PM_(2.5). Methods The rats with asthma were sensitized by intraperitoneal injections of ovalbumin and challenged by aerosol inhalation of ovalbumin,then were administrated intratracheally with 3 mg/kg PM_(2.5),and 1.8,7.2 mg/kg PM_(2.5) water soluble components and 0.6,2.4 mg/kg PM_(2.5) organic components once every three days for five times.The pathological changes were observed by HE staining of lung tissue pathological sections. The ratio of Treg cells in peripheral blood were detected by flow cytometry. The levels of IL-10 and TGF-β in bronchoalveolar lavage fluid(BALF) were determined by ELISA. Results Compared with saline control group,asthma group,asthma+PM_(2.5) group and asthma+1.8 mg/kg PM_(2.5) water soluble extracts group,the ratio of Treg cells in peripheral blood was significantly decreased in asthma+7.2 mg/kg PM_(2.5) water soluble extracts group(P<0.05);The ratio of Treg cells in peripheral blood of asthma + 1.8 mg/kg PM_(2.5) organic extracts group was significantly decreased compared with that in asthma + DMSO group and asthma + PM_(2.5) group(P<0.05). Compared with saline control group,asthma group and asthma + PM_(2.5) group,the level of IL-10 in BALF in asthma+7.2 mg/kg PM_(2.5) water soluble extracts group was significantly increased(P<0.05);The level of IL-10 in BALF in asthma+0.6 mg/kg PM_(2.5) organic extracts group was decreased compared with that in asthma + PM_(2.5) group(P<0.05). The level of TGF-β1 in BALF in asthma +PM_(2.5) group was significantly higher than those in asthma group and asthma+1.8 mg/kg PM_(2.5) water soluble extracts group(P<0.05);The levels of TGF-β1 in BALF in asthma + PM_(2.5) group,asthma + 0.6 mg/kg PM_(2.5) organic extracts group and asthma+2.4 mg/kg PM_(2.5) organic extracts group was increased compared with that in asthma + DMSO group(P<0.05). Conclusion The water soluble extracts and organic extracts of traffic-related PM_(2.5) can suppress the proliferation and differentiation of Treg,and cause the changes of corresponding cytokines to aggravate asthma.
引文
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[17]张佳琪，郑鑫，张志红，等.miR155和STAT6在大气PM2.5加重大鼠哮喘中的作用[J].环境与职业医学，2017,34(3):239-244.
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[21]钟巨斌，林千文，谭建新，等.PM2.5对哮喘急性期小鼠脾脏CD4+T淋巴细胞分化的影响[J].中华实用儿科临床杂志，2015,30(16):1269-1272.
[22]Moore KW,De WMR,Coffman RL,et al.Interleukin-10 and the interleukin-10 receptor[J].Annu Rev Immunol,2001,19:683-765.
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[24]Kearley J,Barker JE,Robinson DS,et al.Resolution of airway inflammation and hyperreactivity after in vivo transfer of CD4+CD25+regulatory T cells is interleukin 10 dependent[J].J Exper Med,2005,202:1539.
[25]Annette MC,Alan JR,Stephen PP,et al.Kinetics of IL-10production after segmental antigen challenge of atopic asthmatic subjects[J].J Aller Clin Immunol,2000,106:880-886.
[26]Zhang X,Zhong W,Meng Q,et al.Ambient PM2.5exposure exacerbates severity of allergic asthma in previously sensitized mice[J].J Asthma,2015,52:785-794.
[27]徐佳莉，王莹，陈鹏．高剂量PM2.5诱导卵清蛋白致哮喘小鼠肺损伤及其机制[J]．细胞与分子免疫学杂志，2017,33(10):1297-1302.
[28]Duvernelle C,Freund V,Frossard N.Transforming growth factor-βand its role in asthma[J].Pulmonary Pharmacol Therap,2003,16:181-196.
[29]Bommireddy R,Doetschman T.TGFβ1 and Treg cells:alliance for tolerance[J].Trend Mol Med,2007,13:492-501.
[30]Redington AE,Madden J,Frew AJ,et al.Transforming growth factor-beta 1 in asthma:measurement in bronchoalveolar lavage fluid.[J].Am J Resp Critical Care Med,1997,156:642-647.
[31]赵志旭，冯学斌，石涛，等.哮喘大鼠淋巴液与血液CD4+CD25+Treg及IL-10、TGF-β水平比较和地塞米松干预的影响[J].细胞与分子免疫学杂志，2010,26(3):238-241.
[2]Sram RJ,Binkova B,Dostal M,et al.Health impact of air pollution to children.[J].Int J Hyg Environ Health,2013,216:533-540.
[3]Brauer M,Avila-Casado C,Fortoul T I,et al.Air pollution and retained particles in the lung[J].Environ Health Perspect,2001,109:1039-1043.
[4]Xing YF,Xu YH,Shi MH,et al.The impact of PM2.5on the human respiratory system[J].J Thorac Dis,2016,8:E69.
[5]Pui DYH,Chen SC,Zuo Z.PM2.5,in China:measurements,sources,visibility and health effects,and mitigation[J].Particuology,2014,13:1-26.
[6]Sinclair AH,Melly S,Tolsma D,et al.Childhood asthma acute primary care visits,traffic,and traffic-related pollutants[J].J Air Waste Manag Assoc,2014,64:561-567.
[7]Wagner JG,Morishita M,Keeler GJ,et al.Divergent effects of urban particulate air pollution on allergic airway responses in experimental asthma:a comparison of field exposure studies[J].Environ Health,2012,11:45.
[8]郭潇繁，王汉宁，李典.大气颗粒物诱发哮喘的免疫调控机制研究进展[J].国外医学（卫生学分册），2008(5):264-267.
[9]杨凌，马琼锦，李莉珊，等.PM2.5亚慢性染毒对小鼠肺部炎症及Th17/Treg的影响[J].卫生研究，2014,43(3):387-392.
[10]张志红，杨文敏.汽油车排出颗粒物的化学组分分析[J].中国公共卫生，2001,17(7):623-624.
[11]郑鑫，张佳琪，张志红，等.iNOS基因甲基化对交通相关PM2.5诱导大鼠哮喘加重中NO的调控作用[J].环境与健康杂志，2016,33(11):941-945.
[12]Jacquemin B,Siroux V,Sanchez M,et al.Ambient air pollution and adult asthma incidence in Six European cohorts(ESCAPE)[J].Environ Health Perspect,2015,123:613-621.
[13]Belleudi V,Faustini A,Stafoggia M,et al.Impact of fine and ultrafine particles on emergency hospital admissions for cardiac and respiratory diseases[J].Epidemiology,2010,21:414.
[14]韩建彪，张志红，童国强，等.太原市大气PM2.5对哮喘患者炎症因子的影响[J].环境与健康杂志，2014,31(3):229-231.
[15]闫庆倩，赵学彬，杨莉，等.不同地区大气PM2.5致大鼠肺损伤的比较实验研究[J].环境与健康杂志，2012,29(1):7-11.
[16]Chowdhury PH,Okano H,Honda A,et al.Aqueous and organic extract of PM2.5,collected in different seasons and cities of Japan differently affect respiratory and immune systems[J].Environ Pollut,2017,235:223.
[17]张佳琪，郑鑫，张志红，等.miR155和STAT6在大气PM2.5加重大鼠哮喘中的作用[J].环境与职业医学，2017,34(3):239-244.
[18]Vila J,Isaacs JD,Anderson AE.Regulatory T cells and autoimmunity[J].Curr Opinion Hematol,2009,38:274-279.
[19]Xu W,Lan Q,Chen M,et al.Adoptive transfer of induced-Treg cells effectively attenuates murine airway allergic inflammation[J].PloSOne,2012,7:e40314.
[20]施宇衡，时国朝，万欢英，等．支气管哮喘患者外周血Th17、CD4+CD25+Treg细胞表达特征[J]．中国免疫学杂志，2010,26(8):740-743.
[21]钟巨斌，林千文，谭建新，等.PM2.5对哮喘急性期小鼠脾脏CD4+T淋巴细胞分化的影响[J].中华实用儿科临床杂志，2015,30(16):1269-1272.
[22]Moore KW,De WMR,Coffman RL,et al.Interleukin-10 and the interleukin-10 receptor[J].Annu Rev Immunol,2001,19:683-765.
[23]Saraiva M,O'Garra A.The regulation of IL-10 production by immune cells[J].Nat Rev Immunol,2010,10:170-81.
[24]Kearley J,Barker JE,Robinson DS,et al.Resolution of airway inflammation and hyperreactivity after in vivo transfer of CD4+CD25+regulatory T cells is interleukin 10 dependent[J].J Exper Med,2005,202:1539.
[25]Annette MC,Alan JR,Stephen PP,et al.Kinetics of IL-10production after segmental antigen challenge of atopic asthmatic subjects[J].J Aller Clin Immunol,2000,106:880-886.
[26]Zhang X,Zhong W,Meng Q,et al.Ambient PM2.5exposure exacerbates severity of allergic asthma in previously sensitized mice[J].J Asthma,2015,52:785-794.
[27]徐佳莉，王莹，陈鹏．高剂量PM2.5诱导卵清蛋白致哮喘小鼠肺损伤及其机制[J]．细胞与分子免疫学杂志，2017,33(10):1297-1302.
[28]Duvernelle C,Freund V,Frossard N.Transforming growth factor-βand its role in asthma[J].Pulmonary Pharmacol Therap,2003,16:181-196.
[29]Bommireddy R,Doetschman T.TGFβ1 and Treg cells:alliance for tolerance[J].Trend Mol Med,2007,13:492-501.
[30]Redington AE,Madden J,Frew AJ,et al.Transforming growth factor-beta 1 in asthma:measurement in bronchoalveolar lavage fluid.[J].Am J Resp Critical Care Med,1997,156:642-647.
[31]赵志旭，冯学斌，石涛，等.哮喘大鼠淋巴液与血液CD4+CD25+Treg及IL-10、TGF-β水平比较和地塞米松干预的影响[J].细胞与分子免疫学杂志，2010,26(3):238-241.
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