Full Text:   <565>

Summary:  <130>

CLC number: X511

On-line Access: 2018-06-04

Received: 2017-02-24

Revision Accepted: 2017-06-16

Crosschecked: 2018-05-14

Cited: 0

Clicked: 1203

Citations:  Bibtex RefMan EndNote GB/T7714

 ORCID:

De-zhao Lu

https://orcid.org/0000-0001-5227-6445

-   Go to

Article info.
Open peer comments

Journal of Zhejiang University SCIENCE B 2018 Vol.19 No.6 P.458-470

10.1631/jzus.B1700103


Proteomic analysis of human umbilical vein endothelial cells exposed to PM2.5


Author(s):  Ji Zhu, Lin-wen-si Zhu, Jin-huan Yang, Ying-ling Xu, Cui Wang, Zhuo-yu Li, Wei Mao, De-zhao Lu

Affiliation(s):  Clinical Laboratory, the Third Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou 310053, China; more

Corresponding email(s):   ludezhao@126.com

Key Words:  Fine ambient particulate matter (PM2.5), Human umbilical vein endothelial cell (HUVEC), Proteomics, Toxic mechanism


Ji Zhu, Lin-wen-si Zhu, Jin-huan Yang, Ying-ling Xu, Cui Wang, Zhuo-yu Li, Wei Mao, De-zhao Lu. Proteomic analysis of human umbilical vein endothelial cells exposed to PM2.5[J]. Journal of Zhejiang University Science B, 2018, 19(6): 458-470.

@article{title="Proteomic analysis of human umbilical vein endothelial cells exposed to PM2.5",
author="Ji Zhu, Lin-wen-si Zhu, Jin-huan Yang, Ying-ling Xu, Cui Wang, Zhuo-yu Li, Wei Mao, De-zhao Lu",
journal="Journal of Zhejiang University Science B",
volume="19",
number="6",
pages="458-470",
year="2018",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.B1700103"
}

%0 Journal Article
%T Proteomic analysis of human umbilical vein endothelial cells exposed to PM2.5
%A Ji Zhu
%A Lin-wen-si Zhu
%A Jin-huan Yang
%A Ying-ling Xu
%A Cui Wang
%A Zhuo-yu Li
%A Wei Mao
%A De-zhao Lu
%J Journal of Zhejiang University SCIENCE B
%V 19
%N 6
%P 458-470
%@ 1673-1581
%D 2018
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B1700103

TY - JOUR
T1 - Proteomic analysis of human umbilical vein endothelial cells exposed to PM2.5
A1 - Ji Zhu
A1 - Lin-wen-si Zhu
A1 - Jin-huan Yang
A1 - Ying-ling Xu
A1 - Cui Wang
A1 - Zhuo-yu Li
A1 - Wei Mao
A1 - De-zhao Lu
J0 - Journal of Zhejiang University Science B
VL - 19
IS - 6
SP - 458
EP - 470
%@ 1673-1581
Y1 - 2018
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B1700103


Abstract: 
Exposure to fine ambient particulate matter (PM2.5) is known to be associated with cardiovascular disease. To uncover the molecular mechanisms involved in cardiovascular toxicity of PM2.5, we investigated alterations in the protein profile of human umbilical vein endothelial cells (HUVECs) treated with PM2.5 using two-dimensional electrophoresis in conjunction with mass spectrometry (MS). A total of 31 protein spots were selected as differentially expressed proteins and identified by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) MS. The results demonstrated that DNA damage and cell apoptosis are important factors contributing to PM2.5-mediated toxicity in HUVECs. It is further proposed that PM2.5 can inhibit superoxide dismutase (SOD) activity and increase reactive oxygen species (ROS) and malonaldehyde (MDA) production in a concentration-dependent manner. Induction of apoptosis and DNA damage through oxidative stress pathways may be one of the key toxicological events occurring in HUVECs under PM2.5 stress. These results indicated that the toxic mechanisms of PM2.5 on cardiovascular disease are related to endothelial dysfunction.

大气细颗粒物PM2.5对人脐静脉内皮细胞蛋白质组的影响

目的:应用双向电泳及质谱技术分析PM2.5对人脐静脉内皮细胞(HUVEC)蛋白质组的影响,探讨PM2.5引起心血管毒性的潜在机制.
创新点:多项流行病学调查表明,PM2.5与人类心血管疾病有密切关系,而血管内皮细胞是血管的第一道防线.本项目应用蛋白质组技术研究PM2.5对血管内皮细胞损伤的作用,并从DNA损伤的角度探讨了PM2.5心血管毒性机制,具有较好的创新性.研究结果可为PM2.5的危害分析提供基础数据,同时为PM2.5的防治提供新的依据及思路.
方法:培养HUVEC细胞,分为正常组(正常培养的HUVEC细胞)、PM2.5处理组(50、100 µg/ml PM2.5处理HUVEC细胞24 h).双向电泳技术建立各组细胞蛋白质组图谱,质谱技术鉴定差异表达的蛋白质,流式细胞术分析细胞凋亡情况,酶联免疫吸附法(ELISA)检测DNA损伤产物8-OHdG的水平,荧光标记技术分析DNA双链断裂的形成,并用免疫印迹法(Western blotting)检测DNA损伤相关蛋白的表达.
结论:经PM2.5处理HUVEC后,31个蛋白表达发生了显著性的变化(图2,表1),其中8个蛋白质参与了DNA的损伤与RNA的编缉,7个蛋白质与细胞凋亡有关(表2).进一步实验表明:PM2.5能够促细胞凋亡(图5),提高DNA损伤产物8-OHdG的含量(图6),促进DNA双链断裂位点的形成(图7),调节损伤修复相关蛋白(Mer11A、Rad50和Rad51)的表达(图8),抑制超氧化物歧化酶(SOD)的活性,增加HUVEC细胞内活性氧(ROS)和丙二醛(MDA)的水平(图9).综上所述,PM2.5能够通过调节一系列蛋白的表达,加重HUVEC细胞氧化应激水平,增加DNA损伤,促进细胞凋亡,造成内皮细胞损伤,从而导致心脑血管事件的发生.

关键词:PM2.5;人脐静脉内皮细胞;蛋白质组;毒性机制

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article

Reference

[1]Alaoui-Jamali MA, Xu YJ, 2006. Proteomic technology for biomarker profiling in cancer: an update. J Zhejiang Univ-Sci B, 7(6):411-420.

[2]Azad MB, Chen YQ, Gibson SB, 2009. Regulation of autophagy by reactive oxygen species (ROS):implications for cancer progression and treatment. Antioxid Redox Sign, 11(4):777-790.

[3]Brigelius-Flohe R, 2009. Commentary: oxidative stress reconsidered. Genes Nutr, 4(3):161-163.

[4]Brook RD, Rajagopalan S, Pope CA, et al., 2010. Particulate matter air pollution and cardiovascular disease: an update to the scientific statement from the American Heart Association. Circulation, 121(21):2331-2378.

[5]Burnett RT, Smith-Doiron M, Stieb D, et al., 1999. Effects of particulate and gaseous air pollution on cardiorespiratory hospitalizations. Arch Environ Health, 54(2):130-139.

[6]Calderón-Garcidueñas L, Villarreal-Calderon R, Valencia-Salazar G, et al., 2008. Systemic inflammation, endothelial dysfunction, and activation in clinically healthy children exposed to air pollutants. Inhal Toxicol, 20(5):499-506.

[7]Cheng Q, Yang CY, Guo BY, et al., 2017. Analysis of mechanism of PM2.5 and house dust mite antigen Der p1 in attack stage of child asthma. Eur Rev Med Pharmacol Sci, 21(10):2458-2462.

[8]Garneau D, Revil T, Fisette JF, et al., 2005. Heterogeneous nuclear ribonucleoprotein F/H proteins modulate the alternative splicing of the apoptotic mediator Bcl-x. J Biol Chem, 280(24):22641-22650.

[9]Ghio AJ, Carraway MS, Madden MC, 2012. Composition of air pollution particles and oxidative stress in cells, tissues, and living systems. J Toxicol Environ Health Part B, 15(1):1-21.

[10]Hirano S, Furuyama A, Koike E, et al., 2003. Oxidative-stress potency of organic extracts of diesel exhaust and urban fine particles in rat heart microvessel endothelial cells. Toxicology, 187(2-3):161-170.

[11]Lee BJ, Kim B, Lee K, 2014. Air pollution exposure and cardiovascular disease. Toxicol Res, 30(2):71-75.

[12]Lee IT, Yang CM, 2012. Role of NADPH oxidase/ROS in pro-inflammatory mediators-induced airway and pulmonary diseases. Biochem Pharmacol, 84(5):581-590.

[13]Li N, Hao MQ, Phalen RF, et al., 2003. Particulate air pollutants and asthma: a paradigm for the role of oxidative stress in PM-induced adverse health effects. Clin Immunol, 109(3):250-265.

[14]Li RJ, Kou XJ, Geng H, et al., 2014. Pollution characteristics of ambient PM2.5-bound PAHs and NPAHs in a typical winter time period in Taiyuan. Chin Chem Lett, 25(5):663-666.

[15]Lu H, Daugherty A, 2013. Atherosclerosis: cell biology and lipoproteins. Curr Opin Lipidol, 24(1):107-108.

[16]Nicolas J, Chiari M, Crespo J, et al., 2008. Quantification of Saharan and local dust impact in an arid Mediterranean area by the positive matrix factorization (PMF) technique. Atmos Environ, 42(39):8872-8882.

[17]Pope CA, Burnctt RT, Thurston GD, et al., 2004. Cardiovascular mortality and long-term exposure to particulate air pollution: epidemiological evidence of general pathophysiological pathways of disease. Circulation, 109(1):71-77.

[18]Rundell KW, Hoffman JR, Caviston R, et al., 2007. Inhalation of ultrafine and fine particulate matter disrupts systemic vascular function. Inhal Toxicol, 19(2):133-140.

[19]Singh R, Sharma BS, 2012. Composition, seasonal variation, and sources of PM10 from world heritage site Taj Mahal, Agra. Environ Monit Assess, 184(10):5945-5956.

[20]Terzano C, Di Stefano F, Conti V, et al., 2010. Air pollution ultrafine particles: toxicity beyond the lung. Eur Rev Med Pharmacol Sci, 14(10):809-821.

[21]Torres-Ramos YD, Montoya-Estrada A, Guzman-Grenfell AM, et al., 2011. Urban PM2.5 induces ROS generation and RBC damage in COPD patients. Front Biosci, 3:808-817.

[22]Ucker DS, Jain MR, Pattabiraman G, et al., 2012. Externalized glycolytic enzymes are novel, conserved, and early biomarkers of apoptosis. J Biol Chem, 287(13):10325-10343.

[23]Vallejo M, Ruiz S, Hermosillo AG, et al., 2006. Ambient fine particles modify heart rate variability in young healthy adults. J Expo Sci Environ Epidemiol, 16(2):125-130.

[24]Wang JS, Tseng CY, Chao MW, 2017. Diesel exhaust particles contribute to endothelia apoptosis via autophagy pathway. Toxicol Sci, 156(1):72-83.

[25]Weichenthal SA, Godri-Pollitt K, Villeneuve PJ, 2013. PM2.5, oxidant defence and cardiorespiratory health: a review. Environ Health, 12:40.

[26]Zhang HH, Li Z, Liu Y, et al., 2018. Physical and chemical characteristics of PM2.5 and its toxicity to human bronchial cells BEAS-2B in the winter and summer. J Zhejiang Univ-Sci B (Biomed & Biotechnol), 19(4):317-326.

Open peer comments: Debate/Discuss/Question/Opinion

<1>

Please provide your name, email address and a comment





Journal of Zhejiang University-SCIENCE, 38 Zheda Road, Hangzhou 310027, China
Tel: +86-571-87952783; E-mail: cjzhang@zju.edu.cn
Copyright © 2000 - Journal of Zhejiang University-SCIENCE