Environmental Science and Pollution Research

, Volume 26, Issue 9, pp 8758–8767 | Cite as

Sulfur dioxide induces apoptosis via reactive oxygen species generation in rat cardiomyocytes

  • Shuyue Li
  • Zhifang Xu
  • Jin Xia
  • Guohua QinEmail author
  • Nan SangEmail author
Research Article


Epidemiological evidence suggests that the incidence and mortality of cardiovascular diseases are closely related to sulfur dioxide (SO2). In the present study, H9C2 cells were incubated with 100 μM NaHSO3 with or without pretreatment of an antioxidant, N-acetyl-l-cysteine (NAC). The changes of apoptosis rate, mitochondrial membrane potential (MMP), ATP content, caspase-3 activity, and reactive oxygen species (ROS) were detected. Rats were inhaled 7 mg/m3 SO2 and/or intraperitoneal injected with 50 mg/kg (bw) of NAC for 30 days. RT-PCR and Western blot were used to detect the mRNA and protein levels of apoptosis-related genes. We found that the apoptosis of H9C2 cells was induced by NaHSO3, which decreased the content of MMP and ATP, and induced the expression of caspase-3. NAC can inhibit the apoptosis induced by NaHSO3 treatment. SO2 and NaHSO3 decreased the expression of Bcl-2 and the ratio of Bcl-2/Bax, increased the expression of Bax and P53 accumulation and phosphorylation, and activated caspase-9 and caspase-3. Whereas NAC can reduce the changes of apoptosis-related proteins in rat heart. Our results suggest that SO2 induces ROS-mediated P53 and caspase-dependent mitochondrial signaling pathways in H9C2 cells and rat hearts. Antioxidant therapy can reduce the adverse reactions of SO2 and lead to a decline in the cardiovascular disease induced by SO2.


Sulfur dioxide Apoptosis Hearts Antioxidant Reactive oxygen species 



This research was supported by the National Natural Science Foundation of China (21777091, 21377076, 91543203, 21222701), Research Project for Young Sanjin Scholarship of Shanxi, Program for the Outstanding Innovative Teams of Higher Learning Institutions of Shanxi, State Key Laboratory of Environmental Chemistry and Ecotoxicology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences (KF2016-17), and Research Project Supported by Shanxi Scholarship Council of China (015-006).


  1. AlMatar M, Batool T, Makky EA (2016) Therapeutic potential of N-acetylcysteine for wound healing, acute bronchiolitis, and congenital heart defects. Curr Drug Metab 17(2):156–167CrossRefGoogle Scholar
  2. Bai J, Meng Z (2005a) Effects of sulfur dioxide on apoptosis-related gene expressions in lungs from rats. Regul Toxicol Pharmacol 43:272–279CrossRefGoogle Scholar
  3. Bai J, Meng Z (2005b) Expression of apoptosis-related genes in livers from rats exposed to sulfur dioxide. Toxicology 216:253–260CrossRefGoogle Scholar
  4. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254CrossRefGoogle Scholar
  5. Brook RD, Kousha T (2015) Air pollution and emergency department visits for hypertension in Edmonton and Calgary, Canada: a case-crossover study. Am J Hypertens 28:1121–1126CrossRefGoogle Scholar
  6. Chung JW, Bang OY, Ahn K, Park SS, Park TH, Kim JG, Ko Y, Lee S, Lee KB, Lee J, Kang K, Park JM, Cho YJ, Hong KS, Nah HW, Kim DH, Cha JK, Ryu WS, Kim DE, Kim JT, Choi JC, Oh MS, Yu KH, Lee BC, Lee JS, Lee J, Park HK, Kim BJ, Han MK, Bae HJ (2017) Air pollution is associated with ischemic stroke via cardiogenic embolism. Stroke 48:17–23CrossRefGoogle Scholar
  7. Crowder RN, El-Deiry WS (2012) Caspase-8 regulation of trail-mediated cell death. Exp Oncol 34:160–164Google Scholar
  8. Elmore S (2007) Apoptosis: a review of programmed cell death. Toxicol Pathol 35:495–516CrossRefGoogle Scholar
  9. Green DR, Reed JC (1998) Mitochondria and apoptosis. Science 281:1309–1312CrossRefGoogle Scholar
  10. Hetts SW (1998) To die or not to die: an overview of apoptosis and its role in disease. JAMA 279:300–307CrossRefGoogle Scholar
  11. Hoek G, Brunekreef B, Fischer P, van Wijnen J (2001) The association between air pollution and heart failure, arrhythmia, embolism, thrombosis, and other cardiovascular causes of death in a time series study. Epidemiology 12:355–357CrossRefGoogle Scholar
  12. Ibe W, Saraste A, Lindemann S, Bruder S, Buerke M, Darius H, Pulkki K, Voipio-Pulkki LM (2007) Cardiomyocyte apoptosis is related to left ventricular dysfunction and remodelling in dilated cardiomyopathy, but is not affected by growth hormone treatment. Eur J Heart Fail 9:160–167CrossRefGoogle Scholar
  13. Ji AJ, Savon SR, Jacobsen DW (1995) Determination of total serum sulfite by HPLC with fluorescence detection. Clin Chem 41:897–903Google Scholar
  14. Joseph B, Marchetti P, Formstecher P, Kroemer G, Lewensohn R, Zhivotovsky B (2002) Mitochondrial dysfunction is an essential step for killing of non-small cell lung carcinomas resistant to conventional treatment. Oncogene 21:65–77CrossRefGoogle Scholar
  15. Kang SH, Heo J, Oh IY, Kim J, Lim WH, Cho Y, Choi EK, Yi SM, Do Shin S, Kim H, Oh S (2016) Ambient air pollution and out-of-hospital cardiac arrest. Int J Cardiol 203:1086–1092CrossRefGoogle Scholar
  16. Kubli DA, Zhang X, Lee Y, Hanna RA, Quinsay MN, Nguyen CK, Jimenez R, Petrosyan S, Murphy AN, Gustafsson AB (2013) Parkin protein deficiency exacerbates cardiac injury and reduces survival following myocardial infarction. J Biol Chem 288:915–926CrossRefGoogle Scholar
  17. Labbe’ P, Pelletier M, Omara FO, Girard D (1998) Functional responses of human neutrophils to sodium sulfite (Na2SO3) in vitro. Hum Exp Toxicol 17:600–605CrossRefGoogle Scholar
  18. Li G, Sang N (2009) Delayed rectifier potassium channels are involved in SO2 derivative-induced hippocampal neuronal injury. Ecotoxicol Environ Saf 72:236–241CrossRefGoogle Scholar
  19. Liu Y, Chen X, Huang S, Tian L, Lu Y, Mei Y, Ren M, Li N, Liu L, Xiang H (2015) Association between air pollutants and cardiovascular disease mortality in Wuhan, China. Int J Environ Res Public Health 12:3506–3516CrossRefGoogle Scholar
  20. Meng Z, Liu Y (2007) Cell morphological ultrastructural changes in various organs from mice exposed by inhalation to sulfur dioxide. Inhal Toxicol 19:543–551CrossRefGoogle Scholar
  21. Miyashita T, Krajewski S, Krajewska M, Wang HG, Lin HK, Liebermann DA, Hoffman B, Reed JC (1994) Tumor suppressor p53 is a regulator of bcl-2 and bax gene expression in vitro and in vivo. Oncogene 9:1799–1805Google Scholar
  22. Mottley C, Mason RP, Chignell CF, Sivarajah K, Eling TE (1982) The formation of sulfur trioxide radical anion during the prostaglandin hydroperoxidase-catalyzed oxidation of bisulfite (hydrated sulfur dioxide). J Biol Chem 257:5050–5055Google Scholar
  23. Murriel CL, Churchill E, Inagaki K, Szweda LI, Mochly-Rosen D (2004) Protein kinase activation induces apoptosis in response to cardiac ischemia and reperfusion damage: a mechanism involving BAD and the mitochondria. J Biol Chem 279:47985–47991CrossRefGoogle Scholar
  24. Qin G, Wang J, Huo Y, Yan H, Jiang C, Zhou J, Wang X, Sang N (2012) Sulfur dioxide inhalation stimulated mitochondrial biogenesis in rat brains. Toxicology 300:67–74CrossRefGoogle Scholar
  25. Qin G, Wu M, Sang N (2015) Sulfur dioxide and benzo(a) pyrene trigger apoptotic and anti-apoptotic signals at different post-exposure times in mouse liver. Chemosphere 139:318–325CrossRefGoogle Scholar
  26. Qin G, Wu M, Wang J, Xu Z, Xia J, Sang N (2016) Sulfur dioxide contributes to the cardiac and mitochondrial dysfunction in rats. Toxicol Sci 151:334–346CrossRefGoogle Scholar
  27. Ranguelova K, Rice AB, Khajo A, Triquigneaux M, Garantziotis S, Magliozzo RS, Mason RP (2012) Formation of reactive sulfite- derived free radicals by the activation of human neutrophils: an ESR study. Free Radic Biol Med 52:1264–1271CrossRefGoogle Scholar
  28. Ryter SW, Mizumura K, Choi AM (2014) The impact of autophagy on cell death modalities. Int J Cell Biol 2014:502676CrossRefGoogle Scholar
  29. Soberanes S, Urich D, Baker CM, Burgess Z, Chiarella SE, Bell EL, Ghio AJ, De Vizcaya-Ruiz A, Liu J, Ridge KM, Kamp DW, Chandel NS, Schumacker PT, Mutlu GM, Budinger GR (2009) Mitochondrial complex III-generated oxidants activate ASK1 and JNK to induce alveolar epithelial cell death following exposure to particulate matter air pollution. J Biol Chem 284:2176–2186CrossRefGoogle Scholar
  30. Sunyer J, Ballester F, Tertre AL, Atkinson R, Ayres JG, Forastiere F, Forsberg B, Vonk JM, Bisanti L, Tenias JM, Medina S, Schwartz J, Katsouyanni K (2003) The association of daily sulfur dioxide air pollution levels with hospital admissions for cardiovascular diseases in Europe (the Aphea-II study). Eur Heart J 24:752–760CrossRefGoogle Scholar
  31. Talasaz AH, Khalili H, Fahimi F (2014) Effects of N-acetylcysteine on the cardiac remodeling biomarkers and major adverse events following acute myocardial infarction: a randomized clinical trial. Am J Cardiovasc Drugs 14(1):51–61CrossRefGoogle Scholar
  32. Velayutham M, Hemann CF, Cardounel AJ, Zweier JL (2016) Sulfite oxidase activity of cytochrome c: role of hydrogen peroxide. Biochem Biophys Rep 5:96–104Google Scholar
  33. Wang X, Hu W, Tong S (2009) Long-term exposure to gaseous air pollutants and cardio-respiratory mortality in Brisbane, Australia. Geospat Health 3:257–263CrossRefGoogle Scholar
  34. Wang D, Jiang G, Zhang H, Song G, Zhang Y (2013) Effect of air pollution on coronary heart disease mortality in Tianjin, 2001-2009: a time-series study. Zhonghua Liu Xing Bing Xue Za Zhi 34:478–483Google Scholar
  35. Yang B, Ye D, Wang Y (2013a) Caspase-3 as a therapeutic target for heart failure. Expert Opin Ther Targets 17:255–263CrossRefGoogle Scholar
  36. Yang J, Maity B, Huang J, Gao Z, Stewart A, Weiss RM, Anderson ME, Fisher RA (2013b) G-protein inactivator RGS6 mediates myocardial cell apoptosis and cardiomyopathy caused by doxorubicin. Cancer Res 73:1662–1667CrossRefGoogle Scholar
  37. Yang M, Bo W, Gao J, Yang Z, Xu W, Tao L (2017) Spinosad induces programmed cell death involves mitochondrial dysfunction and cytochrome c release in spodoptera frugiperda, sf9 cells. Chemosphere 169:155–161CrossRefGoogle Scholar
  38. Yun Y, Li H, Li G, Sang N (2010) SO2 inhalation modulates the expression of apoptosis-related genes in rat hippocampus via its derivatives in vivo. Inhal Toxicol 22:919–929CrossRefGoogle Scholar
  39. Zhao X, Jin HF, Tang CS, Du JB (2008) Effects of sulfur dioxide, on the proliferation and apoptosis of aorta smooth muscle cells in hypertension: experiments with rats. Zhonghua Yi Xue Za Zhi 88:1279–1283Google Scholar
  40. Zhao A, Chen R, Kuang X, Kan H (2014) Ambient air pollution and daily outpatient visits for cardiac arrhythmia in Shanghai, China. J Epidemiol 24:321–326CrossRefGoogle Scholar

Copyright information

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

Authors and Affiliations

  1. 1.College of Environment and Resource, Research Center of Environment and HealthShanxi UniversityTaiyuanPeople’s Republic of China
  2. 2.Shan Xi Academy for Environmental PlanningTaiyuanPeople’s Republic of China

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