PM_(2.5)对小鼠和H9C2细胞心肌肥大相关因子mRNA表达的影响
|
摘要
用采自太原市4个不同季节的细颗粒物(PM_(2.5))对小鼠心肌细胞H9C2进行染毒后,采用荧光定量PCR技术检测心肌肥大相关因子ANP和TGF-β的mRNA水平.采用浓度分别为0、1、3、10μg·mL~(-1)的冬季PM_(2.5)处理H9C2细胞后,检测心肌肥大相关因子ANP、β-MHC、MMP2和MMP9的mRNA水平.之后分别选取4周龄、4月龄、10月龄C57BL/6雌性小鼠作为实验模型,采用咽后壁滴注的方法用3 mg·kg~(-1) PM_(2.5)暴露4周.采用苏木素-伊红染色(H&E)对小鼠心脏组织病理切片进行观察,并检测ANP和β-MHC的mRNA水平.结果发现,与对照组相比,冬季PM_(2.5)诱导H9C2细胞中ANP和TGF-β的mRNA水平升高最为显著;其中,当冬季PM_(2.5)染毒浓度较高时可诱导ANP、β-MHC、MMP2和MMP9的mRNA水平均显著升高.经PM_(2.5)暴露4周后,幼年和老年小鼠心肌细胞核质比显著降低.老年小鼠心脏组织中ANP和β-MHC的mRNA表达水平显著上升.实验表明,冬季PM_(2.5)诱导心肌肥大标志物表达改变的效应要强于其他季节,且有一定的剂量效应关系,而老年小鼠对PM_(2.5)诱导的心肌肥大效应最为敏感.
H9C2 cells were exposed to fine particulate matters(PM_(2.5)) from four different seasons in Taiyuan City, the mRNA levels of ANP and TGF-β related to cardiac hypertrophy were detected by real-time PCR. H9C2 cells were exposed to winter PM_(2.5) at a concentration of 0, 1, 3, and 10 μg·mL~(-1), the mRNA levels of ANP, β-MHC, MMP2, and MMP9 were measured by real-time PCR. Female C57 BL/6 mice at 4-week, 4-month, and 10-month were exposed to 3 mg·kg~(-1) PM_(2.5) for 4 weeks by the mesns of posterior pharyngeal wall drip method. The histological sections of heart were observed by hematoxylin-eosin staining(H&E); the mRNA levels of ANP and β-MHC were detected. The results showed that the mRNA levels of ANP and TGF-β in H9C2 cells were significantly increased after exposed to winter PM_(2.5) compared with the control group. Winter PM at higher concentrations could increase the mRNA levels of ANP, β-MHC, MMP2 and MMP9. After four weeks exposure, the nucleoplasmic ratios of cardiomyocytes in young and older mice were significantly reduced.The mRNA expression levels of ANP and β-MHC in cardiac tissues of older mice were significantly increased. The study indicated that the effect of the winter PM_(2.5) increased cardiac hypertrophy markers expression is stronger than other seasons,with a dose-effect manner. While older mice are the most sensitive to PM_(2.5)-induced cardiac hypertrophy.
H9C2 cells were exposed to fine particulate matters(PM_(2.5)) from four different seasons in Taiyuan City, the mRNA levels of ANP and TGF-β related to cardiac hypertrophy were detected by real-time PCR. H9C2 cells were exposed to winter PM_(2.5) at a concentration of 0, 1, 3, and 10 μg·mL~(-1), the mRNA levels of ANP, β-MHC, MMP2, and MMP9 were measured by real-time PCR. Female C57 BL/6 mice at 4-week, 4-month, and 10-month were exposed to 3 mg·kg~(-1) PM_(2.5) for 4 weeks by the mesns of posterior pharyngeal wall drip method. The histological sections of heart were observed by hematoxylin-eosin staining(H&E); the mRNA levels of ANP and β-MHC were detected. The results showed that the mRNA levels of ANP and TGF-β in H9C2 cells were significantly increased after exposed to winter PM_(2.5) compared with the control group. Winter PM at higher concentrations could increase the mRNA levels of ANP, β-MHC, MMP2 and MMP9. After four weeks exposure, the nucleoplasmic ratios of cardiomyocytes in young and older mice were significantly reduced.The mRNA expression levels of ANP and β-MHC in cardiac tissues of older mice were significantly increased. The study indicated that the effect of the winter PM_(2.5) increased cardiac hypertrophy markers expression is stronger than other seasons,with a dose-effect manner. While older mice are the most sensitive to PM_(2.5)-induced cardiac hypertrophy.
引文
Bell M L,Ebisu K,Peng R D,et al.2008.Seasonal and regional short-term effects of fine particles on hospital admissions in 202 US counties,1999-2005[J].American Journal of Epidemiology,168(11):1301-1310
Brook R D,Rajagopalan S,Pope C A,et al.2010.Particulate matter air pollution and cardiovascular disease an update to the scientific statement from the american heart association[J].Circulation,121(21):2331-2378
Cuspidi C,Negri F,Sala C,et al.2012.Age related prevalence of severe left ventricular hypertrophy in essential hypertension: echocardiographic findings from the ETODH study[J].Blood Press,21(3):139-145
Chen L H,Knutsen S F,Shavlik D,et al.2005.The association between fatal coronary heart disease and ambient particulate air pollution: Are females at greater risk?[J].Environmental Health Perspectives,113(12):1723-1729
Dockery D W,Xu X,Spengler J D,et al.1993.An association between air pollution and mortality in six U.S.cities[J].New England Journal of Medicine,329(24):1753-1759
董晨,曹强,宋伟民.2006.大气PM2.5对自发性高血压大鼠心脏的急性毒作用[J].环境与职业医学,23(2):96-99
Fananapazir L,Dalakas M C,Cyran F,et al.1993.Missense mutations in the beta-myosin heavy-chain gene cause central core disease in hypertrophic cardiomyopathy[J].Proceedings of the National Academy of Sciences of the United States of America, 90(9):3993, DOI:10.1073/pnas.90.9.3993
Galis Z S,Sukhova G K,Lark M W,et al.1994.Increased expression of matrix metalloproteinases and matrix degrading activity in vulnerable regions of human atherosclerotic plaques[J].Journal of Clinical Investigation,94(6):2493-2503
Hunter J J,Chien K R.1999.Signaling pathways for cardiac hypertrophy and failure[J].The New Englang Journal of Medicine,341(17):1276-1283
贺擎,丁文军,芮魏,等.2012.冬季和夏季PM10和PM2.5人肺上皮细胞A549毒性的比较[J].中国科学院大学学报, 29(3):324-331
Kan H,London S J,Chen G,et al.2008.Season,sex,age,and education as modifiers of the effects of outdoor air pollution on daily mortality in shanghai,China: The public health and air pollution in asia (PAPA) study[J].Environmental Health Perspectives,116(9):1183-1188
刘英,廖玉华,程翔,等.2005.急性心肌梗死后心脏TGF-β1的表达和心室重塑[J].中国病理生理杂志,21(12):2305-2309
Liu Y,Chien W M,Medvedev I O,et al.2013.Inhalation of diesel exhaust does not exacerbate cardiac hypertrophy or heart failure in two mouse models of cardiac hypertrophy[J].Particle & Fibre Toxicology,10(1):49
Malemud C J.2006.Matrix metalloproteinases (MMPs) in health and disease: An overview[J].Front Biosci,11(2):1696-1701
Pope R C,Burnett R T,Thurston G D,et al.2004.Cardiovascular mortality and long-term exposure to particulate air pollution: epidemiological evidence of general pathophysiological pathways of disease[J].Circulation,109(1):71-77
Pandya K,Kim H S,Smithies O.2006.Fibrosis,not cell size,delineates β-myosin heavy chain reexpression during cardiac hypertrophy and normal aging in vivo[J].Proceedings of the National Academy of Sciences of the United States of America,103(45):16864-16869
Qin G H,Xia J,Zhang Y Y,et al.2018.Ambient fine particulate matter exposure induces reversible cardiac dysfunction and fibrosis in juvenile and older female mice[J].Particle and Fibre Toxicology,15:27
Rooij E V,Sutherland L B,Liu N,et al.2006.A signature pattern of stress-responsive microRNAs that can evoke cardiac hypertrophy and heart failure[J].Proceedings of the National Academy of Sciences of the United States of America,103(48):18255-18260
任艳军,李秀央,金明娟,等.2006.大气颗粒物污染与心血管疾病死亡的病例交叉研究[D].杭州:浙江大学
Spinale F G.2002.Matrix metalloproteinases regulation and dysregulation in the failing heart[J].Circulation Research, 90(5):520-530
Sacks J D,Stanek L W,Luben T J,et al.2011.Particulate matter-induced health effects: Who is susceptible?[J].Environmental Health Perspectives,119(4):446-454
Tao J,Gao J,Zhang L,et al.2014.Pm2.5 pollution in a megacity of southwest China: Source apportionment and implication[J].Atmospheric Chemistry & Physics,14(4):8679-8699
Wilson W,Claiborn C S, Hemming B L.2004.Air quality criteria for particulate matter[R].EPA 600/P-99/002aF-bF 2004.Washington,D.C: U.S.Environmental Protection Agency
王欣,邓芙蓉,吴少伟,等.2010.北京市某区大气可吸入颗粒物和细颗粒物对儿童肺功能的短期影响[J].北京大学学报(医学版),42(3):340-344
Younes A,Boluyt M O,O′Neill L,et al.1995.Age-associated increase in rat ventricular ANP gene expression correlates with cardiac hypertrophy[J].American Journal of Physiology,269(3 Pt 2):H1003
Ying Z,Yue P,Xu X,et al.2009.Air pollution and cardiac remodeling: a role for RhoA/Rho-kinase[J].American Journal of Physiology,296(1):H1540
Zhang Y,Ji X,Ku T,et al.2016.Heavy metals bound to fine particulate matter from northern China induce season-dependent health risks: A study based on myocardial toxicity[J].Environmental Pollution,216:380-390
Brook R D,Rajagopalan S,Pope C A,et al.2010.Particulate matter air pollution and cardiovascular disease an update to the scientific statement from the american heart association[J].Circulation,121(21):2331-2378
Cuspidi C,Negri F,Sala C,et al.2012.Age related prevalence of severe left ventricular hypertrophy in essential hypertension: echocardiographic findings from the ETODH study[J].Blood Press,21(3):139-145
Chen L H,Knutsen S F,Shavlik D,et al.2005.The association between fatal coronary heart disease and ambient particulate air pollution: Are females at greater risk?[J].Environmental Health Perspectives,113(12):1723-1729
Dockery D W,Xu X,Spengler J D,et al.1993.An association between air pollution and mortality in six U.S.cities[J].New England Journal of Medicine,329(24):1753-1759
董晨,曹强,宋伟民.2006.大气PM2.5对自发性高血压大鼠心脏的急性毒作用[J].环境与职业医学,23(2):96-99
Fananapazir L,Dalakas M C,Cyran F,et al.1993.Missense mutations in the beta-myosin heavy-chain gene cause central core disease in hypertrophic cardiomyopathy[J].Proceedings of the National Academy of Sciences of the United States of America, 90(9):3993, DOI:10.1073/pnas.90.9.3993
Galis Z S,Sukhova G K,Lark M W,et al.1994.Increased expression of matrix metalloproteinases and matrix degrading activity in vulnerable regions of human atherosclerotic plaques[J].Journal of Clinical Investigation,94(6):2493-2503
Hunter J J,Chien K R.1999.Signaling pathways for cardiac hypertrophy and failure[J].The New Englang Journal of Medicine,341(17):1276-1283
贺擎,丁文军,芮魏,等.2012.冬季和夏季PM10和PM2.5人肺上皮细胞A549毒性的比较[J].中国科学院大学学报, 29(3):324-331
Kan H,London S J,Chen G,et al.2008.Season,sex,age,and education as modifiers of the effects of outdoor air pollution on daily mortality in shanghai,China: The public health and air pollution in asia (PAPA) study[J].Environmental Health Perspectives,116(9):1183-1188
刘英,廖玉华,程翔,等.2005.急性心肌梗死后心脏TGF-β1的表达和心室重塑[J].中国病理生理杂志,21(12):2305-2309
Liu Y,Chien W M,Medvedev I O,et al.2013.Inhalation of diesel exhaust does not exacerbate cardiac hypertrophy or heart failure in two mouse models of cardiac hypertrophy[J].Particle & Fibre Toxicology,10(1):49
Malemud C J.2006.Matrix metalloproteinases (MMPs) in health and disease: An overview[J].Front Biosci,11(2):1696-1701
Pope R C,Burnett R T,Thurston G D,et al.2004.Cardiovascular mortality and long-term exposure to particulate air pollution: epidemiological evidence of general pathophysiological pathways of disease[J].Circulation,109(1):71-77
Pandya K,Kim H S,Smithies O.2006.Fibrosis,not cell size,delineates β-myosin heavy chain reexpression during cardiac hypertrophy and normal aging in vivo[J].Proceedings of the National Academy of Sciences of the United States of America,103(45):16864-16869
Qin G H,Xia J,Zhang Y Y,et al.2018.Ambient fine particulate matter exposure induces reversible cardiac dysfunction and fibrosis in juvenile and older female mice[J].Particle and Fibre Toxicology,15:27
Rooij E V,Sutherland L B,Liu N,et al.2006.A signature pattern of stress-responsive microRNAs that can evoke cardiac hypertrophy and heart failure[J].Proceedings of the National Academy of Sciences of the United States of America,103(48):18255-18260
任艳军,李秀央,金明娟,等.2006.大气颗粒物污染与心血管疾病死亡的病例交叉研究[D].杭州:浙江大学
Spinale F G.2002.Matrix metalloproteinases regulation and dysregulation in the failing heart[J].Circulation Research, 90(5):520-530
Sacks J D,Stanek L W,Luben T J,et al.2011.Particulate matter-induced health effects: Who is susceptible?[J].Environmental Health Perspectives,119(4):446-454
Tao J,Gao J,Zhang L,et al.2014.Pm2.5 pollution in a megacity of southwest China: Source apportionment and implication[J].Atmospheric Chemistry & Physics,14(4):8679-8699
Wilson W,Claiborn C S, Hemming B L.2004.Air quality criteria for particulate matter[R].EPA 600/P-99/002aF-bF 2004.Washington,D.C: U.S.Environmental Protection Agency
王欣,邓芙蓉,吴少伟,等.2010.北京市某区大气可吸入颗粒物和细颗粒物对儿童肺功能的短期影响[J].北京大学学报(医学版),42(3):340-344
Younes A,Boluyt M O,O′Neill L,et al.1995.Age-associated increase in rat ventricular ANP gene expression correlates with cardiac hypertrophy[J].American Journal of Physiology,269(3 Pt 2):H1003
Ying Z,Yue P,Xu X,et al.2009.Air pollution and cardiac remodeling: a role for RhoA/Rho-kinase[J].American Journal of Physiology,296(1):H1540
Zhang Y,Ji X,Ku T,et al.2016.Heavy metals bound to fine particulate matter from northern China induce season-dependent health risks: A study based on myocardial toxicity[J].Environmental Pollution,216:380-390