Archives of Toxicology

, Volume 93, Issue 4, pp 1009–1020 | Cite as

Particulate matter 2.5 induced arrhythmogenesis mediated by TRPC3 in human induced pluripotent stem cell-derived cardiomyocytes

  • Cheng Cai
  • Jiayi Huang
  • Yongping Lin
  • Weilun Miao
  • Pei Chen
  • Xing Chen
  • Jiaxian WangEmail author
  • Minglong ChenEmail author
Organ Toxicity and Mechanisms


Particulate matter (PM) is one of the most important environmental issues worldwide, which is associated with not only pulmonary but also cardiovascular diseases. However, the underlying biological mechanisms of PM related cardiovascular dysfunction remained poorly defined, especially mediated by the pathway of direct impact on vascular and heart. Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) provide an ideal platform for studying PM-exposed cellular diseases model in vitro. Here, we investigated the direct effects of particulate matter 2.5 (PM2.5) on hiPSC-CMs and the potential mechanism at non-cytotoxic concentrations. Cell viability, contraction amplitude and spontaneous beating rate of iPSC-CMs after direct exposure to PM2.5 showed that the concentration of lower than 100 µg/ml would not lead to cytotoxic effects. Calcium-mediated optical mapping illustrated that there was a concentration-dependent reduction in quantification of calcium transient amplitude and an increase in the incidence of early after depolarizations due to PM2.5 treatment. Furthermore, there were dramatic dosage-dependent shortening in action potential duration and decrease in L-type calcium peak current density. The Ingenuity Pathway Analysis of our transcriptive study indicated that PM2.5 exposure preferentially influenced the expression of genes involved in calcium signaling. Among them the up-regulation of TRPC3 potentially played an important role in the electrophysiological alteration of PM2.5 on hiPSC-CMs, which could be ameliorated by pretreatment with pyr3, the inhibitor of TRPC3. In conclusion, our results demonstrated that exposure to PM2.5 was capable of increasing propensity to cardiac arrhythmias which could be attenuated with TRPC3 inhibition.


Particulate matter Human induced pluripotent stem cells Cardiomyocytes Electrophysiology TRPC3 



Particulate matter


Cardiovascular disease


Particulate matter 2.5


Human induced pluripotent stem cell


Human induced pluripotent stem cell-derived cardiomyocytes


Phosphate-buffered saline


Cell counting kit


CardioExcyte 96


Cardiac troponin-I


Early afterdepolarization


Action potential


Real-time polymerase chain reaction


Action potential duration


Ingenuity pathway analysis


Diesel exhaust particulate


Compliance with ethical standards

Conflict of interest

The authors declared that they have no conflict of interest.

Ethical standards

The manuscript does not contain clinical studies or patient data.

Supplementary material

204_2019_2403_MOESM1_ESM.pdf (469 kb)
Supplementary material 1 (PDF 469 KB)


  1. Bagate K, Meiring JJ, Cassee FR, Borm PJ (2004) The effect of particulate matter on resistance and conductance vessels in the rat. Inhal Toxicol 16(6–7):431–436. CrossRefGoogle Scholar
  2. Brook RD, Franklin B, Cascio W et al (2004) Air pollution and cardiovascular disease: a statement for healthcare professionals from the Expert Panel on Population and Prevention Science of the American Heart Association. Circulation 109(21):2655–2671. CrossRefGoogle Scholar
  3. 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. CrossRefGoogle Scholar
  4. Brookes PS, Yoon Y, Robotham JL, Anders MW, Sheu SS (2004) Calcium, ATP, and ROS: a mitochondrial love-hate triangle. Am J Physiol Cell Physiol 287(4):C817–C833. CrossRefGoogle Scholar
  5. Burridge PW, Keller G, Gold JD, Wu JC (2012) Production of de novo cardiomyocytes: human pluripotent stem cell differentiation and direct reprogramming. Cell Stem Cell 10(1):16–28. CrossRefGoogle Scholar
  6. Cao J, Qin G, Shi R et al (2016) Overproduction of reactive oxygen species and activation of MAPKs are involved in apoptosis induced by PM2.5 in rat cardiac H9c2 cells. J Appl Toxicol 36(4):609–617. CrossRefGoogle Scholar
  7. Carr MJ, Undem BJ (2001) Inflammation-induced plasticity of the afferent innervation of the airways. Environ Health Perspect 109(Suppl 4):567–571Google Scholar
  8. Cohen AJ, Brauer M, Burnett R et al (2017) Estimates and 25-year trends of the global burden of disease attributable to ambient air pollution: an analysis of data from the Global Burden of Diseases Study 2015. Lancet 389(10082):1907–1918. CrossRefGoogle Scholar
  9. Dockery DW, Luttmann-Gibson H, Rich DQ et al (2005) Association of air pollution with increased incidence of ventricular tachyarrhythmias recorded by implanted cardioverter defibrillators. Environ Health Perspect 113(6):670–674. CrossRefGoogle Scholar
  10. Feng S, Li H, Tai Y et al (2013) Canonical transient receptor potential 3 channels regulate mitochondrial calcium uptake. Proc Natl Acad Sci USA 110(27):11011–11016. CrossRefGoogle Scholar
  11. Fuks KB, Weinmayr G, Basagana X et al (2017) Long-term exposure to ambient air pollution and traffic noise and incident hypertension in seven cohorts of the European study of cohorts for air pollution effects (ESCAPE). Eur Heart J 38(13):983–990. Google Scholar
  12. Furuyama A, Kanno S, Kobayashi T, Hirano S (2009) Extrapulmonary translocation of intratracheally instilled fine and ultrafine particles via direct and alveolar macrophage-associated routes. Arch Toxicol 83(5):429–437. CrossRefGoogle Scholar
  13. Gao H, Wang F, Wang W et al (2012) Ca(2+) influx through L-type Ca(2+) channels and transient receptor potential channels activates pathological hypertrophy signaling. J Mol Cell Cardiol 53(5):657–667. CrossRefGoogle Scholar
  14. Geng L, Wang Z, Cui C et al (2018) Rapid electrical stimulation increased cardiac apoptosis through disturbance of calcium homeostasis and mitochondrial dysfunction in human induced pluripotent stem cell-derived cardiomyocytes. Cell Physiol Biochem 47(3):1167–1180. CrossRefGoogle Scholar
  15. Gorr MW, Youtz DJ, Eichenseer CM et al (2015) In vitro particulate matter exposure causes direct and lung-mediated indirect effects on cardiomyocyte function. Am J Physiol Heart Circ Physiol 309(1):H53–H62. CrossRefGoogle Scholar
  16. Harada M, Luo X, Qi XY et al (2012) Transient receptor potential canonical-3 channel-dependent fibroblast regulation in atrial fibrillation. Circulation 126(17):2051–2064. CrossRefGoogle Scholar
  17. Hazari MS, Haykal-Coates N, Winsett DW et al (2011) TRPA1 and sympathetic activation contribute to increased risk of triggered cardiac arrhythmias in hypertensive rats exposed to diesel exhaust. Environ Health Perspect 119(7):951–957. CrossRefGoogle Scholar
  18. Helms AS, Alvarado FJ, Yob J et al (2016) Genotype-Dependent and -independent calcium signaling dysregulation in human hypertrophic cardiomyopathy. Circulation 134(22):1738–1748. CrossRefGoogle Scholar
  19. Ide T, Tsutsui H, Hayashidani S et al (2001) Mitochondrial DNA damage and dysfunction associated with oxidative stress in failing hearts after myocardial infarction. Circ Res 88(5):529–535CrossRefGoogle Scholar
  20. January CT, Riddle JM (1989) Early afterdepolarizations: mechanism of induction and block. A role for L-type Ca2+ current. Circ Res 64(5):977–990CrossRefGoogle Scholar
  21. Kim JB, Kim C, Choi E et al (2012) Particulate air pollution induces arrhythmia via oxidative stress and calcium calmodulin kinase II activation. Toxicol Appl Pharmacol 259(1):66–73. CrossRefGoogle Scholar
  22. Kiyonaka S, Kato K, Nishida M et al (2009) Selective and direct inhibition of TRPC3 channels underlies biological activities of a pyrazole compound. Proc Natl Acad Sci U S A 106(13):5400–5405. CrossRefGoogle Scholar
  23. Lelieveld J, Evans JS, Fnais M, Giannadaki D, Pozzer A (2015) The contribution of outdoor air pollution sources to premature mortality on a global scale. Nature 525(7569):367–371. CrossRefGoogle Scholar
  24. Link MS, Luttmann-Gibson H, Schwartz J et al (2013) Acute exposure to air pollution triggers atrial fibrillation. J Am Coll Cardiol 62(9):816–825. CrossRefGoogle Scholar
  25. Ljungman PL, Berglind N, Holmgren C et al (2008) Rapid effects of air pollution on ventricular arrhythmias. Eur Heart J 29(23):2894–2901. CrossRefGoogle Scholar
  26. Lombardi R, Bell A, Senthil V et al (2008) Differential interactions of thin filament proteins in two cardiac troponin T mouse models of hypertrophic and dilated cardiomyopathies. Cardiovasc Res 79(1):109–117. CrossRefGoogle Scholar
  27. Madhvani RV, Xie Y, Pantazis A et al (2011) Shaping a new Ca2+ conductance to suppress early afterdepolarizations in cardiac myocytes. J Physiol 589(Pt 24):6081–6092. CrossRefGoogle Scholar
  28. Makarewich CA, Zhang H, Davis J et al (2014) Transient receptor potential channels contribute to pathological structural and functional remodeling after myocardial infarction. Circ Res 115(6):567–580. CrossRefGoogle Scholar
  29. Matsa E, Burridge PW, Yu KH et al (2016) Transcriptome Profiling of patient-specific human iPSC-cardiomyocytes predicts individual drug safety and efficacy responses in vitro. Cell Stem Cell 19(3):311–325. CrossRefGoogle Scholar
  30. Mills NL, Tornqvist H, Gonzalez MC et al (2007) Ischemic and thrombotic effects of dilute diesel-exhaust inhalation in men with coronary heart disease. N Engl J Med 357(11):1075–1082. CrossRefGoogle Scholar
  31. Mitcheson JS, Hancox JC, Levi AJ (1998) Cultured adult cardiac myocytes: future applications, culture methods, morphological and electrophysiological properties. Cardiovasc Res 39(2):280–300CrossRefGoogle Scholar
  32. Molkentin JD, Lu JR, Antos CL et al (1998) A calcineurin-dependent transcriptional pathway for cardiac hypertrophy. Cell 93(2):215–228CrossRefGoogle Scholar
  33. Nakayama H, Wilkin BJ, Bodi I, Molkentin JD (2006) Calcineurin-dependent cardiomyopathy is activated by TRPC in the adult mouse heart. FASEB J 20(10):1660–1670. CrossRefGoogle Scholar
  34. Nay Aung M, Mihir MRCP, Sanghvi M, BSc M, Filip Zemrak M et al (2018) Association between ambient air pollution and cardiac morpho-functional phenotypes insights from the UK Biobank Population Imaging Study. Circulation. Google Scholar
  35. Nemmar A, Hoet PH, Vanquickenborne B et al (2002) Passage of inhaled particles into the blood circulation in humans. Circulation 105(4):411–414CrossRefGoogle Scholar
  36. Oberdorster G, Sharp Z, Atudorei V et al (2002) Extrapulmonary translocation of ultrafine carbon particles following whole-body inhalation exposure of rats. J Toxicol Environ Health A 65(20):1531–1543. CrossRefGoogle Scholar
  37. Onohara N, Nishida M, Inoue R et al (2006) TRPC3 and TRPC6 are essential for angiotensin II-induced cardiac hypertrophy. EMBO J 25(22):5305–5316. CrossRefGoogle Scholar
  38. Patel C, Yan GX, Antzelevitch C (2010) Short QT syndrome: from bench to bedside. Circ Arrhythm Electrophysiol 3(4):401–408. CrossRefGoogle Scholar
  39. Pope CA, Burnett 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. CrossRefGoogle Scholar
  40. Pope CA, Bhatnagar A, McCracken JP, Abplanalp W, Conklin DJ, O’Toole T (2016) Exposure to fine particulate air pollution is associated with endothelial injury and systemic inflammation. Circ Res 119(11):1204–1214. CrossRefGoogle Scholar
  41. Rich DQ, Mittleman MA, Link MS et al (2006) Increased risk of paroxysmal atrial fibrillation episodes associated with acute increases in ambient air pollution. Environ Health Perspect 114(1):120–123. CrossRefGoogle Scholar
  42. Sabourin J, Robin E, Raddatz E (2011) A key role of TRPC channels in the regulation of electromechanical activity of the developing heart. Cardiovasc Res 92(2):226–236. CrossRefGoogle Scholar
  43. Sayed N, Liu C, Wu JC (2016) Translation of human-induced pluripotent stem cells: from clinical trial in a dish to precision medicine. J Am Coll Cardiol 67(18):2161–2176. CrossRefGoogle Scholar
  44. Sun Q, Yue P, Kirk RI et al (2008) Ambient air particulate matter exposure and tissue factor expression in atherosclerosis. Inhal Toxicol 20(2):127–137. CrossRefGoogle Scholar
  45. Suwa T, Hogg JC, Quinlan KB, Ohgami A, Vincent R, van Eeden SF (2002) Particulate air pollution induces progression of atherosclerosis. J Am Coll Cardiol 39(6):935–942CrossRefGoogle Scholar
  46. Tamagawa E, Bai N, Morimoto K et al (2008) Particulate matter exposure induces persistent lung inflammation and endothelial dysfunction. Am J Physiol Lung Cell Mol Physiol 295(1):L79–L85. CrossRefGoogle Scholar
  47. Tse G, Yan BP, Chan YW, Tian XY, Huang Y (2016) Reactive oxygen species, endoplasmic reticulum stress and mitochondrial dysfunction: the link with cardiac arrhythmogenesis. Front Physiol 7:313. Google Scholar
  48. Watkinson WP, Campen MJ, Nolan JP, Costa DL (2001) Cardiovascular and systemic responses to inhaled pollutants in rodents: effects of ozone and particulate matter. Environ Health Perspect 109(Suppl 4):539–546CrossRefGoogle Scholar
  49. Wold LE, Ying Z, Hutchinson KR et al (2012) Cardiovascular remodeling in response to long-term exposure to fine particulate matter air pollution. Circ Heart Fail 5(4):452–461. CrossRefGoogle Scholar
  50. Yang J, Rothermel B, Vega RB et al (2000) Independent signals control expression of the calcineurin inhibitory proteins MCIP1 and MCIP2 in striated muscles. Circ Res 87(12):E61–E68CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Division of CardiologyThe First Affiliated Hospital of Nanjing Medical UniversityNanjingChina
  2. 2.HELP Stem Cell InnovationsNanjingChina

Personalised recommendations