Skip to main content
SpringerLink
Log in

Sodium Fluoride Induces Apoptosis in H9c2 Cardiomyocytes by Altering Mitochondrial Membrane Potential and Intracellular ROS Level

  • Published:
Biological Trace Element Research Aims and scope Submit manuscript

Abstract

Chronic excessive fluoride intake is known to be toxic, and effects of long-term fluorosis on different organ systems have been examined. However, there are few studies about the effects of fluorosis on cardiovascular systems. Here, we studied the fluoride-induced apoptosis in H9c2 cells and determined the underlying molecular mechanisms including the cell viability, intracellular reactive oxygen species (ROS) level, the changes of mitochondrial membrane potential (ΔΨm), and the cell apoptosis. Sodium fluoride (NaF) at concentrations of 0, 2, 4, 8, and 16 mg/L was administered to cultured H9c2 cells for up to 48 h. After the treatment, H9c2 cells were collected and the associated parameters were measured by flow cytometry. Our study found that fluoride not only inhibited H9c2 cell proliferation but also induced cell apoptosis. With the increment of NaF concentration, the apoptotic rates and ROS generation were increased, while the ΔΨm was decreased. In summary, these data suggested that NaF-induced H9c2 cell apoptosis is mediated by direct increased intracellular ROS and downregulated ΔΨm.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Aguirre-Sierra A, Alonso A, Camargo JA (2013) Fluoride bioaccumulation and toxic effects on the survival and behavior of the endangered white-clawed crayfish Austropotamobius pallipes (Lereboullet). Arch Environ Contam Toxicol 65:244–250

    Article  CAS  PubMed  Google Scholar 

  2. Mukhopadhyay D, Chattopadhyay A (2014) Induction of oxidative stress and related transcriptional effects of sodium fluoride in female zebrafish liver. Bull Environ Contam Toxicol 93:64–70

    Article  CAS  PubMed  Google Scholar 

  3. Adali MK, Varol E, Aksoy F, Icli A, Ersoy IH, Ozaydin M, Erdogan D, Dogan A (2013) Impaired heart rate recovery in patients with endemic fluorosis. Biol Trace Elem Res 152:310–315

    Article  CAS  PubMed  Google Scholar 

  4. Trevors JT, Saier MH Jr (2009) Where is the global environmental bailout? Water Air Soil Pollut 198:1–3

    Article  CAS  Google Scholar 

  5. Ayoob S, Gupta A (2006) Fluoride in drinking water: a review on the status and stress effects. Crit Rev Environ Sci Technol 55:433–487

    Article  Google Scholar 

  6. Wang Y, Gu ZW (1996) Modern occupational medicine. People's medical publishing house, Beijing

    Google Scholar 

  7. Cicek E, Aydin G, Akdogan M, Okutan H (2005) Effects of chronic ingestion of sodium fluoride on myocardium in a second generation of rats. Hum Exp Toxicol 24:79–87

    Article  CAS  PubMed  Google Scholar 

  8. Basha MP, Sujitha NS (2011) Chronic fluoride toxicity and myocardial damage: antioxidant offered protection in second generation rats. Toxicol Int 18:99–104

    Article  PubMed Central  PubMed  Google Scholar 

  9. Zhan WC, He DJ (1988) Ultrastructural findings in liver, kidneys, thyroid gland and myocardium of rabbits following sodium fluoride administration. J Guiyang Med Coll 13:129–132

    Google Scholar 

  10. Bian J, Lin X, Yang X, Fan T, Zhu Q (2010) Changes of certain oxidative, anti-oxidative and vascular function indexes of New Zealand rabbit exposed by high-fluoride. Wei Sheng Yan Jiu 39:751–754

    CAS  PubMed  Google Scholar 

  11. Shen XY, Ruan Q, Zhang ZG, Xu XL (2003) Influence of fluorosis on hemodynamics of rats. Chin J Control Endemic Dis 18:208–209

    Google Scholar 

  12. Cheng RY, Nie QL, Sun HF, Zhang YJ, Wu LH, Ma YQ, Yan XY (2013) Fluoride-induced oxidative stress in rat myocardium through the Bax/Bcl-2 signalling pathway. Fluoride 46(4):198–203

    CAS  Google Scholar 

  13. Sinha M, Manna P, Sil PC (2008) Terminalia arjuna protects mouse hearts against sodium fluoride-induced oxidative stress. J Med Food 11:733–740

    Article  CAS  PubMed  Google Scholar 

  14. Ly JD, Grubb DR, Lawen A (2003) The mitochondrial membrane potential (∆ψm) in apoptosis; an update. Apoptosis 8:115–128

    Article  CAS  PubMed  Google Scholar 

  15. Kroemer G, Zazami N, Susin SA (1997) Immunol Today 18:44–51

    Article  CAS  PubMed  Google Scholar 

  16. Green DR, Reed JC (1998) Mitochondria and apoptosis. Science 281:1309–1312

    Article  CAS  PubMed  Google Scholar 

  17. Kowaltowski AJ, Netto LE, Vercesi AE (1998) The thiol-specific antioxidant enzyme prevents mitochondrial permeability transition. Evidence for the participation of reactive oxygen species in this mechanism. J Biol Chem 273:12766–12769

    Article  CAS  PubMed  Google Scholar 

  18. Cai J, Jones DP (1998) Superoxide in apoptosis. Mitochondrial generation triggered by cytochrome c loss. J Biol Chem 273:11401–11404

    Article  CAS  PubMed  Google Scholar 

  19. Kim CN, Wang X, Huang Y, Ibrado AM, Liu L, Fang G, Bhalla K (1997) Overexpression of Bcl-X (L) inhibits Ara-C-induced mitochondrial loss of cytochrome C and other perturbations that activate the molecular cascade of apoptosis. Cancer Res 57:3115–3120

    CAS  PubMed  Google Scholar 

  20. Duranteau J, Chandel NS, Kulisz A, Shao Z, Schumacker PT (1998) Intracellular signaling by reactive oxygen species during hypoxia in cardiomyocytes. J Biol Chem 273:11619–11624

    Article  CAS  PubMed  Google Scholar 

  21. Cai L, Li W, Wang G, Guo L, Jiang Y, Kang YJ (2002) Hyperglycemia-induced apoptosis in mouse myocardium: mitochondrial cytochrome c-mediated caspase-3 activation pathway. Diabetes 51:1938–1948

    Article  CAS  PubMed  Google Scholar 

  22. Sun Z, Niu R, Wang B, Jiao Z, Wang J, Zhang J, Wang S, Wang J (2011) Fluoride-induced apoptosis and gene expression profiling in mice sperm in vivo. Arch Toxicol 85:1441–1452

    Article  CAS  PubMed  Google Scholar 

  23. Ge Y, Niu R, Zhang J, Wang J (2011) Proteomic analysis of brain proteins of rats exposed to high fluoride and low iodine. Arch Toxicol 85:27–33

    Article  CAS  PubMed  Google Scholar 

  24. Yan X, Feng C, Chen Q, Li W, Wang H, Lv L, Smith GW, Wang J (2009) Effects of sodium fluoride treatment in vitro on cell proliferation, apoptosis and caspase-3 and caspase-9 mRNA expression by neonatal rat osteoblasts. Arch Toxicol 83:451–458

    Article  CAS  PubMed  Google Scholar 

  25. Dubey N, Khan AM, Raina R (2013) Sub-acute deltamethrin and fluoride toxicity induced hepatic oxidative stress and biochemical alterations in rats. Bull Environ Contam Toxicol 91:334–338

    Article  CAS  PubMed  Google Scholar 

  26. Shivashankara AR, Shivarajashankara YM, Bhat PG, Rao SH (2002) Lipid peroxidation and antioxidant defence systems in liver of rats in chronic fluoride toxicity. Bull Environ Contam Toxicol 68:612–616

    Article  CAS  PubMed  Google Scholar 

  27. Hassan HA, Yousef MI (2009) Mitigating effects of antioxidant properties of black berry juice on sodium fluoride induced hepatotoxicity and oxidative stress in rats. Food Chem Toxicol 47:2332–2337

    Article  CAS  PubMed  Google Scholar 

  28. Ranjan R, Swarup D, Patra RC (2009) Oxidative stress indices in erythrocytes, liver, and kidneys of fluoride-exposed rabbits. Fluoride 42:88–93

    CAS  Google Scholar 

  29. Dönmez N, Çinar A (2003) Effects of chronic fluorosis on electrocardiogram in sheep. Biol Trace Elem Res 92:115–121

    Article  PubMed  Google Scholar 

  30. Miyamoto H, Doita M, Nishida K, Yamamoto T, Sumi M, Kurosaka M (2006) Effects of cyclic mechanical stress on the production of inflammatory agents by nucleus pulposus and anulus fibrosus derived cells in vitro. Spine 31:4–9

    Article  PubMed  Google Scholar 

  31. Sohal RS, Weindruch R (1996) Oxidative stress, caloric restriction, and aging. Science 273:59–63

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  32. Beatty S, Koh H, Phil M et al (2000) The role of oxidative stress in the pathogenesis of age-related macular degeneration. Surv Ophthalmol 45:115–134

    Article  CAS  PubMed  Google Scholar 

  33. Sang H, Zhang L, Li J (2012) Anti-benzopyrene-7, 8-diol-9, 10-epoxide induces apoptosis via mitochondrial pathway in human bronchiolar epithelium cells independent of the mitochondria permeability transition pore. Food Chem Toxicol 50:2417–2423

    Article  CAS  PubMed  Google Scholar 

  34. Khan M, Ding C, Rasul A, Yi F, Li T, Gao H, Gao R, Zhong L, Zhang K, Fang X, Ma T (2012) Isoalantolactone induces reactive oxygen species mediated apoptosis in pancreatic carcinomapanc-1 cells. Int J Biol Sci 8:533–547

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  35. Barbier O, Arreola-Mendoza L, Del Razo LM (2010) Molecular mechanisms of fluoride toxicity. Chem Biol Interact 188:319–333

    Article  CAS  PubMed  Google Scholar 

  36. Carmody RJ, Cotter TG (2001) Signalling apoptosis: a radical approach. Redox Reports 6:77–90

    Article  CAS  Google Scholar 

  37. Chandra J, Samali A, Orrenius S (2000) Triggering and modulation of apoptosis by oxidative stress. Free Radic Biol Med 29:323–333

    Article  CAS  PubMed  Google Scholar 

  38. Shih CM, Ko WC, Wu JS, Wei YH, Wang LF, Chang EE, Lo TY, Cheng HH, Chen CT (2004) Mediating of caspase-independent apoptosis by cadmium through the mitochondria-ROS pathway in MRC-5 fibroblasts. J Cell Biochem 91:384–397

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This research was sponsored by the China National Natural Science Foundation (Grant Nos. 31240009, 31302158); Shanxi Scholarship Council of China (Grant No. 2012–079); Shanxi Province Science and Technology Bureau Program (Grant Nos. 2011021030-1, No. 2013011059-1) and Shanxi Province Soft Science (Grant No. 2013041084-03)

Conflicts of Interest

There are no conflicts of interest.

Author information

Authors and Affiliations

  1. Department of Biology, Taiyuan Normal University, (TYNU), Taiyuan, 030031, People’s Republic of China

    Xiaoyan Yan, Xianhui Hao, Qiurong Ren & Na Chang

  2. Shanxi Key Laboratory of Experimental Animal Science and Animal Model of Human Disease, Health Toxicology Department, School of Public Health, Shanxi Medical University, Taiyuan, 030001, People’s Republic of China

    Xiaoyan Yan, Xia Yang, Jiping Gao, Yu Wang, Yulan Qiu & Guohua Song

Authors
  1. Xiaoyan Yan
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Xia Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Xianhui Hao
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Qiurong Ren
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Jiping Gao
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Yu Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Na Chang
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Yulan Qiu
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Guohua Song
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Xiaoyan Yan or Guohua Song.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Yan, X., Yang, X., Hao, X. et al. Sodium Fluoride Induces Apoptosis in H9c2 Cardiomyocytes by Altering Mitochondrial Membrane Potential and Intracellular ROS Level. Biol Trace Elem Res 166, 210–215 (2015). https://doi.org/10.1007/s12011-015-0273-z

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12011-015-0273-z

Keywords

Search

Navigation