Skip to main content

Thank you for visiting You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Effects of ozone treatment on penile erection capacity and nitric oxide synthase levels in diabetic rats


We aimed to determine the effects of ozone treatment on functional and biochemical changes in corpus cavernosum of diabetic rats. A total of 18 rats were included in the study. The rats were divided into the three groups as control, diabetes mellitus, and diabetes mellitus + ozone therapy groups. In the latter, ozone gas mixture was administered intraperitoneally for 2 weeks after the induction of experimental diabetes model. Erectile response was evaluated by determining mean intracavernosal pressure. Tissue neuronal, inducible and endothelial nitric oxide synthase levels were evaluated with commercial ELISA kits. Immunohistochemical evaluation was also performed to determine the expression levels of nitric oxide synthases semiquantatively. Mean intracavernosal pressure and intracavernosal pressure/systemic arterial blood pressure ratio were significantly higher in the diabetes mellitus + ozone therapy group than those of diabetes mellitus group (24.57 ± 6.36 mmHg vs. 5.98 ± 2.04 mmHg, p = 0.005 and 0.81 ± 0.16 vs. 0.26 ± 0.11, p = 0.0001, respectively). The level of penile tissue endothelial nitric oxide synthase was significantly higher in diabetes mellitus + ozone therapy group compared with others (19.28 ± 3.40 ng/mL vs. 13.47 ± 2.06 ng/mL and 13.28 ± 1.48 ng/mL, P = 0.01). Endothelial nitric oxide synthase expression increased significantly with ozone therapy. Our results suggest that ozone therapy may be beneficial in reducing the negative effects of diabetes on erectile dysfunction as a result of enhanced enzymatic activity in endothelial nitric oxide synthase levels.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Fig. 1: MAP and ICP parameters in the groups.
Fig. 2: Representative ICP recordings of each experimental study group.
Fig. 3: Tissue levels of the parameters by ELISA.
Fig. 4: Tissue eNOS expressions.
Fig. 5: Tissue iNOS expressions.


  1. 1.

    Giugliano F, Maiorino M, Bellastella G, Gicchino M, Giugliano D, Esposito K. Determinants of erectile dysfunction in type 2 diabetes. Int J Impot Res. 2010;22:204–9.

    CAS  PubMed  Article  Google Scholar 

  2. 2.

    Chen S, Peng D, Xu X, Gao J, Dai F, Zuo C, et al. Assessment of erectile dysfunction and associated psychological distress in Chinese men with type 2 diabetes mellitus. Int J Impot Res. 2017;29:210–14.

    CAS  PubMed  Article  Google Scholar 

  3. 3.

    Esposito K, Ciotola M, Giugliano F, Schisano B, Improta L, Improta MR, et al. Endothelial microparticles correlate with erectile dysfunction in diabetic men. Int J Impot Res. 2007;19:161–6.

    CAS  PubMed  Article  Google Scholar 

  4. 4.

    Ignarro LJ, Bush PA, Buga GM, Wood KS, Fukuto JM, Rajfer J. Nitric oxide and cyclic GMP formation upon electrical field stimulation cause relaxation of corpus cavernosum smooth muscle. Biochem Biophys Res Commun. 1990;170:843–50.

    CAS  PubMed  Article  Google Scholar 

  5. 5.

    Musicki B, Burnett AL. Endothelial dysfunction in diabetic erectile dysfunction. Int J Impot Res. 2007;19:129–38.

    CAS  PubMed  Article  Google Scholar 

  6. 6.

    Cashen DE, MacIntyre DE, Martin WJ. Effects of sildenafil on erectile activity in mice lacking neuronal or endothelial nitric oxide synthase. Br J Pharmacol. 2002;136:693–700.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  7. 7.

    Pryor WA. Mechanisms of radical formation from reactions of ozone with target molecules in the lung. Free Radic Biol Med. 1994;17:451–65.

    CAS  PubMed  Article  Google Scholar 

  8. 8.

    Leroy P, Tham A, Wong H, Tenney R, Chen C, Stiner R, et al. Inflammatory and repair pathways induced in human bronchoalveolar lavage cells with ozone inhalation. PloS one 2015;10:e0127283.

    PubMed  PubMed Central  Article  Google Scholar 

  9. 9.

    Bialoszewski D, Kowalewski M. Superficially, longer, intermittent ozone theraphy in the treatment of the chronic, infected wounds. Ortop Traumatol Rehabil. 2003;5:652–8.

    PubMed  Google Scholar 

  10. 10.

    Rao V, Kiran R. Evaluation of correlation between oxidative stress and abnormal lipid profile in coronary artery disease. J Cardiovasc Dis Res. 2011;2:57–60.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  11. 11.

    Coppola L, Giunta R, Verrazzo G, Luongo C, Sammartino A, Vicario C, et al. Influence of ozone on haemoglobin oxygen affinity in type-2 diabetic patients with peripheral vascular disease: in vitro studies. Diabete Metab. 1995;21:252–5.

    CAS  PubMed  Google Scholar 

  12. 12.

    Martínez-Sánchez G, Al-Dalain SM, Menéndez S, Re L, Giuliani A, Candelario-Jalil E, et al. Therapeutic efficacy of ozone in patients with diabetic foot. Eur J Pharmacol. 2005;523:151–61.

    PubMed  Article  Google Scholar 

  13. 13.

    Kushmakov R, Gandhi J, Seyam O, Jiang W, Joshi G, Smith NL, et al. Ozone therapy for diabetic foot. Med Gas Res. 2018;8:111–15.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  14. 14.

    Al-Dalain SM, Martínez G, Candelario-Jalil E, Menéndez S, Re L, Giuliani A, et al. Ozone treatment reduces markers of oxidative and endothelial damage in an experimental diabetes model in rats. Pharmacol Res. 2001;44:391–6.

    CAS  PubMed  Article  Google Scholar 

  15. 15.

    Ajamieh H, Merino N, Candelario-Jalil E, Menéndez S, Martinez-Sanchez G, Re L, et al. Similar protective effect of ischaemic and ozone oxidative preconditionings in liver ischaemia/reperfusion injury. Pharmacol Res. 2002;45:333–39.

    CAS  PubMed  Article  Google Scholar 

  16. 16.

    Peralta C, León OS, Xaus C, Prats N, Jalil EC, Planell ES, et al. Protective effect of ozone treatment on the injury associated with hepatic ischemia-reperfusion: antioxidant-prooxidant balance. Free Radic Res. 1999;31:191–96.

    CAS  PubMed  Article  Google Scholar 

  17. 17.

    Oztosun M, Akgul EO, Cakir E, Cayci T, Uysal B, Ogur R, et al. The effects of medical ozone therapy on renal ischemia/reperfusion injury. Ren Fail. 2012;34:921–5.

    CAS  PubMed  Article  Google Scholar 

  18. 18.

    Quinlan DM, Nelson RJ, Partin AW, Mostwin JL, Walsh PC. The rat as a model for the study of penile erection. J Urol. 1989;141:656–61.

    CAS  PubMed  Article  Google Scholar 

  19. 19.

    Martinez-Pineiro L, Brock G, Trigo-Rocha F, Hsu GL, Lue TF, Tanagho EA. Rat model for the study of penile erection: pharmacologic and electrical-stimulation parameters. Eur Urol. 1994;25:62–70.

    CAS  PubMed  Article  Google Scholar 

  20. 20.

    Ratajczak-Wrona W, Jablonska E, Antonowicz B, Dziemianczyk D, Grabowska SZ. Levels of biological markers of nitric oxide in serum of patients with squamous cell carcinoma of the oral cavity. Int J Oral Sci. 2013;5:141–5.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  21. 21.

    Maas R, Schwedhelm E, Albsmeier J, Boger RH. The pathophysiology of erectile dysfunction related to endothelial dysfunction and mediators of vascular function. Vasc Med. (Lond, Engl) 2002;7:213–25.

    Article  Google Scholar 

  22. 22.

    Andersson KE. Erectile physiological and pathophysiological pathways involved in erectile dysfunction. J Urol. 2003;170:S6–13. discussion S13-4

    PubMed  Article  Google Scholar 

  23. 23.

    Gratzke C, Angulo J, Chitaley K, Dai YT, Kim NN, Paick JS, et al. Anatomy, physiology, and pathophysiology of erectile dysfunction. J Sex Med. 2010;7:445–75.

    CAS  PubMed  Article  Google Scholar 

  24. 24.

    Burnett AL, Ricker DD, Chamness SL, Maguire MP, Crone JK, Bredt DS, et al. Localization of nitric oxide synthase in the reproductive organs of the male rat. Biol Reprod. 1995;52:1–7.

    CAS  PubMed  Article  Google Scholar 

  25. 25.

    Bivalacqua TJ, Usta MF, Champion HC, Adams D, Namara DB, Abdel-Mageed AB, et al. Gene transfer of endothelial nitric oxide synthase partially restores nitric oxide synthesis and erectile function in streptozotocin diabetic rats. J Urol. 2003;169:1911–7.

    CAS  PubMed  Article  Google Scholar 

  26. 26.

    Rajasekaran M, Mondal D, Agrawal K, Chen IL, Hellstrom W, Sikka S. Ex vivo expression of nitric oxide synthase isoforms (eNOS/iNOS) and calmodulin in human penile cavernosal cells. J Urol. 1998;160:2210–5.

    CAS  PubMed  Article  Google Scholar 

  27. 27.

    Tao M, Tasdemir C, Tasdemir S, Shahabi A, Liu G. Penile alterations at early stage of type 1 diabetes in rats. Int Braz J Urol. 2017;43:753–61.

    PubMed  PubMed Central  Article  Google Scholar 

  28. 28.

    Yagihashi S, Mizukami H, Sugimoto K. Mechanism of diabetic neuropathy: where are we now and where to go? J Diabet Investig. 2011;2:18–32.

    CAS  Article  Google Scholar 

  29. 29.

    Musicki B, Hannan JL, Lagoda G, Bivalacqua TJ, Burnett AL. Mechanistic link between erectile dysfunction and systemic endothelial dysfunction in type 2 diabetic rats. Andrology 2016;4:977–83.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  30. 30.

    Munzel T, Daiber A, Ullrich V, Mulsch A. Vascular consequences of endothelial nitric oxide synthase uncoupling for the activity and expression of the soluble guanylyl cyclase and the cGMP-dependent protein kinase. Arterioscler Thromb Vasc Biol. 2005;25:1551–7.

    PubMed  Article  Google Scholar 

  31. 31.

    Forstermann U, Li H. Therapeutic effect of enhancing endothelial nitric oxide synthase (eNOS) expression and preventing eNOS uncoupling. Br J Pharmacol. 2011;164:213–23.

    PubMed  PubMed Central  Article  Google Scholar 

  32. 32.

    Di Filippo C, Trotta MC, Maisto R, Siniscalco D, Luongo M, Mascolo L, et al. Daily Oxygen/O(3) Treatment Reduces Muscular Fatigue and Improves Cardiac Performance in Rats Subjected to Prolonged High Intensity Physical Exercise. Oxid Med Cell Longev. 2015;2015:190640.

    PubMed  PubMed Central  Article  Google Scholar 

  33. 33.

    Di Mauro R, Cantarella G, Bernardini R, Di Rosa M, Barbagallo I, Distefano A et al. The biochemical and pharmacological properties of ozone: the smell of protection in acute and chronic diseases. Int J Mol Sci. 2019;20:634–46.

    PubMed Central  Article  Google Scholar 

  34. 34.

    Saleh S, El-Ridi M, Zalat S, El-Kotb S, Donia S. Additive effect of ozone therapy to insulin in the treatment of diabetic rats. Menoufia Med J. 2014;27:85–92.

    Article  Google Scholar 

  35. 35.

    Bocci V, Zanardi I, Huijberts MS, Travagli V. Diabetes and chronic oxidative stress. A perspective based on the possible usefulness of ozone therapy. Diabet Metab Syndr. 2011;5:45–9.

    Article  Google Scholar 

  36. 36.

    Ajamieh HH, Menéndez S, Martínez-Sánchez G, Candelario-Jalil E, Re L, Giuliani A, et al. Effects of ozone oxidative preconditioning on nitric oxide generation and cellular redox balance in a rat model of hepatic ischaemia-reperfusion. Liver Int. 2004;24:55–62.

    CAS  PubMed  Article  Google Scholar 

  37. 37.

    Smith NL, Wilson AL, Gandhi J, Vatsia S, Khan SA. Ozone therapy: an overview of pharmacodynamics, current research, and clinical utility. Med Gas Res. 2017;7:212–19.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  38. 38.

    Pranata MT. Ozone Therapy in Type 2 Diabetic Erectile Dysfunction (A Preliminary Report) ISMH World Congress 2010. J Men’s Health. 2010;7:319–20.

    Google Scholar 

Download references

Author information



Corresponding author

Correspondence to Aykut Colakerol.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Colakerol, A., Temiz, M.Z., Tavukcu, H.H. et al. Effects of ozone treatment on penile erection capacity and nitric oxide synthase levels in diabetic rats. Int J Impot Res 33, 1–8 (2021).

Download citation

Further reading


Quick links