Skip to main content
SpringerLink
Log in

Silymarin and celecoxib ameliorate experimental varicocele-induced pathogenesis: evidences for oxidative stress and inflammation inhibition

  • Urology - Original Paper
  • Published:
International Urology and Nephrology Aims and scope Submit manuscript

Abstract

Background

The present study was done to investigate the ameliorative effect of silymarin (SMN) and celecoxib (CEL) on varicocele (VCL)-induced detrimental impact in testicular tissue.

Methods

Mature Wistar rats were divided into control and test groups. Following VCL induction, the animals in test group were subdivided into non-treated VCL-induced, SMN-treated (50 mg/kg, orally), CEL-treated (10 mg/kg) and SMN + CEL-treated groups. Following 60 days, testicular total antioxidant capacity (TAC), malondialdehyde (MDA), nitric oxide (NO), superoxide dismutase (SOD), glutathione peroxidase (GSH-px), total thiol molecules (TTM), mRNA and protein levels of COX2 and mRNA level of iNos were analyzed. Moreover, the germinal cells apoptosis and mRNA damage were examined.

Results

Observations revealed that co-administration of SMN and CEL significantly (P < 0.05) up-regulated TAC, SOD, GSH-px and TTM levels and resulted in a remarkable (P < 0.05) reduction in iNos and COX2 expression, NO and MDA contents. The animals in SMN + CEL-treated group exhibited significantly (P < 0.05) lower number of apoptotic cells and cells with mRNA damage per one mm2.

Conclusion

The SMN by up-regulating testicular TAC, SOD, GSH-px and TTM levels and the CEL by inhibiting COX2 and iNos expression as well as NO content could fairly ameliorate the VCL-decreased spermatogenesis.

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
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

References

  1. French DB, Desai NR, Agarwal A (2008) Varicocele repair: Does it still have a role in infertility treatment? Curr Opin Obstet Gynecol 20(3):269–274

    Article  PubMed  Google Scholar 

  2. Naughton CK, Nangia AK, Agarwal A (2001) Pathophysiology of varicoceles in male infertility. Hum Reprod Update 7(5):473–481

    Article  PubMed  CAS  Google Scholar 

  3. Agarwal A, Sharma RK, Desai NR, Prabakaran S, Tavares A, Sabanegh E (2009) Role of oxidative stress in pathogenesis of varicocele and infertility. Urology 73(3):461–469

    Article  PubMed  Google Scholar 

  4. Pasqualotto FF, Braga DP, Figueira RC, Setti AS, Iaconelli A Jr, Borges E Jr (2012) Varicocelectomy does not impact pregnancy outcomes following intracytoplasmic sperm injection procedures. J Androl 33(2):239–243

    Article  PubMed  CAS  Google Scholar 

  5. Aitken RJ, De Iuliis GN, Finnie JM, Hedges A, McLachlan RI (2010) Analysis of the relationships between oxidative stress, DNA damage and sperm vitality in a patient population: development of diagnostic criteria. Hum Reprod 25(10):2415–2426

    Article  PubMed  CAS  Google Scholar 

  6. Walczak-Jedrzejowska R, Wolski JK, Slowikowska-Hilczer J (2013) The role of oxidative stress and antioxidants in male fertility. Cent Eur J Urol 66(1):60–67

    Article  CAS  Google Scholar 

  7. Agarwal A, Hamada A, Esteves SC (2012) Insight into oxidative stress in varicocele-associated male infertility: part 1. Nat Rev Urol 9(12):678–690

    Article  PubMed  CAS  Google Scholar 

  8. Khosravanian N, Razi M, Farokhi F, Khosravanian H (2014) Testosterone and vitamin E administration up-regulated varicocele-reduced Hsp70-2 protein expression and ameliorated biochemical alterations. J Assist Reprod Genet 31(3):341–354

    Article  PubMed  PubMed Central  Google Scholar 

  9. Rouzer CA, Marnett LJ (2009) Cyclooxygenases: structural and functional insights. J Lipid Res 50(Suppl):29–34

    Article  CAS  Google Scholar 

  10. Winnall WR, Ali U, O’Bryan MK, Hirst JJ, Whiley PA, Muir JA, Hedger MP (2007) Constitutive expression of prostaglandin-endoperoxide synthase 2 by somatic and spermatogenic cells is responsible for prostaglandin E2 production in the adult rat testis. Biol Reprod 76(5):759–768

    Article  PubMed  CAS  Google Scholar 

  11. Schell C, Frungieri MB, Albrecht M, Gonzalez-Calvar SI, Kohn FM, Calandra RS, Mayerhofer A (2007) A prostaglandin D2 system in the human testis. Fertil Steril 88(1):233–236

    Article  PubMed  CAS  Google Scholar 

  12. Perrotta I, Santoro M, Guido C, Avena P, Tripepi S, De Amicis F, Gervasi MC, Aquila S (2012) Expression of cyclooxygenase-1 (COX-1) and COX-2 in human male gametes from normal patients, and those with varicocele and diabetes: a potential molecular marker for diagnosing male infertility disorders. J Anat 221(3):209–220

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  13. Kantartzi P, Goulis CD, Goulis G, Papadimas I (2007) Male infertility and varicocele: myths and reality. Hippokratia 11(3):99–104

    PubMed  PubMed Central  CAS  Google Scholar 

  14. Razi M, Malekinejad H (2015) Varicocele-induced infertility in animal models. Int J Fertil Steril 9(2):141–149

    PubMed  PubMed Central  CAS  Google Scholar 

  15. Shiraishi K, Naito K (2007) Nitric oxide produced in the testis is involved in dilatation of the internal spermatic vein that compromises spermatogenesis in infertile men with varicocele. BJU Int 99(5):1086–1090

    Article  PubMed  CAS  Google Scholar 

  16. Sahin Z, Celik-Ozenci C, Akkoyunlu G, Korgun ET, Acar N, Erdogru T, Demir R, Ustunel I (2006) Increased expression of interleukin-1α and interleukin-1β is associated with experimental varicocele. Fertil Steril 85:1265–1275

    Article  PubMed  CAS  Google Scholar 

  17. Ishikawa T, Fujioka H, Ishimura T, Takenaka A, Fujisawa M (2007) Expression of leptin and leptin receptor in the testis of fertile and infertile patients. Andrologia 39(1):22–27

    Article  PubMed  CAS  Google Scholar 

  18. Steinbach G, Lynch PM, Phillips RK, Wallace MH, Hawk E, Gordon GB, Wakabayashi N, Saunders B, Shen Y, Fujimura T, Su LK, Levin B, Godio L, Patterson S, Rodriguez-Bigas MA, Jester SL, King KL, Schumacher M, Abbruzzese J, DuBois RN, Hittelman WN, Zimmerman S, Sherman JW, Kelloff G (2000) The effect of celecoxib, a cyclooxygenase-2 inhibitor, in familial adenomatous polyposis. N Engl J Med 342(26):1946–1952

    Article  PubMed  CAS  Google Scholar 

  19. Wang D, DuBois RN (2013) The role of anti-inflammatory drugs in colorectal cancer. Annu Rev Med 64:131–144

    Article  PubMed  CAS  Google Scholar 

  20. Ninomiya I, Nagai N, Oyama K, Hayashi H, Tajima H, Kitagawa H, Fushida S, Fujimura T, Ohta T (2012) Antitumor and anti-metastatic effects of cyclooxygenase-2 inhibition by celecoxib on human colorectal carcinoma xenografts in nude mouse rectum. Oncol Rep 28(3):777–784

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  21. Pogorelic Z, Sopta M, Jukic M, Nevescanin A, Juric I, Furlan D (2017) Laparoscopic varicocelectomy using polymeric ligating clips and its effect on semen parameters in pediatric population with symptomatic varicocele: A 5-year single surgeon experience. J Lapar Adv Surg Tech 27:1318–1325

    Article  Google Scholar 

  22. Jia R, Cao L, Du J, Xu P, Jeney G, Yin G (2013) The protective effect of silymarin on the carbon tetrachloride (CCl4)-induced liver injury in common carp (Cyprinus carpio). Vitro Cell Dev Biol Anim 49(3):155–161

    Article  CAS  Google Scholar 

  23. Perez HJ, Carrillo SC, Garcia E, Ruiz-Mar G, Perez-Tamayo R, Chavarria A (2014) Neuroprotective effect of silymarin in a MPTP mouse model of Parkinson’s disease. Toxicology 319:38–43

    Article  CAS  Google Scholar 

  24. Kren V, Walterova D (2005) Silybin and silymarin—new effects and applications. Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub 149(1):29–41

    Article  PubMed  CAS  Google Scholar 

  25. Nencini C, Giorgi G, Micheli L (2007) Protective effect of silymarin on oxidative stress in rat brain. Phytomedicine 14(2–3):129–135

    Article  PubMed  CAS  Google Scholar 

  26. El-Shitany NA, El-Haggar S, El-desoky K (2008) Silymarin prevents adriamycin-induced cardiotoxicity and nephrotoxicity in rats. Food Chem Toxicol 46(7):2422–2428

    Article  PubMed  CAS  Google Scholar 

  27. Moshtaghion SM, Malekinejad H, Razi M, Shafie-Irannejad V (2013) Silymarin protects from varicocele-induced damages in testis and improves sperm quality: evidence for E2f1 involvement. Syst Biol Reprod Med 59(5):270–280

    Article  PubMed  CAS  Google Scholar 

  28. Hu PJ, Yu J, Zeng ZR, Leung WK, Lin HL, Tang BD, Bai AH, Sung JJ (2004) Chemoprevention of gastric cancer by celecoxib in rats. Gut 53(2):195–200

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  29. Sofikitis N, Miyagawa I (1993) Experimental models for the study of varicocele. Jpn J Fertil Steril 38:168

    CAS  Google Scholar 

  30. Negoescu A, Lorimier P, Labat-Moleur F, Drouet C, Robert C, Guillermet C, Brambilla C, Brambilla E (1996) In situ apoptotic cell labeling by the TUNEL method: improvement and evaluation on cell preparations. J Histochem Cytochem 44(9):959–968

    Article  PubMed  CAS  Google Scholar 

  31. Darzynkiewicz Z (1990) Differential staining of DNA and RNA in intact cells and isolated cell nuclei with acridine orange. Methods Cell Biol 33:285–298

    Article  PubMed  CAS  Google Scholar 

  32. Pant N, Srivastava SP (2003) Testicular and spermatotoxic effects of quinalphos in rats. J Appl Toxicol 23(4):271–274

    Article  PubMed  CAS  Google Scholar 

  33. Niehaus WG Jr, Samuelsson B (1968) Formation of malonaldehyde from phospholipid arachidonate during microsomal lipid peroxidation. Eur J Biochem 6(1):126–130

    Article  PubMed  CAS  Google Scholar 

  34. Malekinejad H, Mirzakhani N, Razi M, Cheraghi H, Alizadeh A, Dardmeh F (2011) Protective effects of melatonin and Glycyrrhiza glabra extract on ochratoxin A-induced damages on testes in mature rats. Hum Exp Toxicol 30(2):110–123

    Article  PubMed  CAS  Google Scholar 

  35. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193(1):265–275

    PubMed  CAS  Google Scholar 

  36. Cheng H, Wang S, Jo YI, Hao CM, Zhang M, Fan X, Kennedy C, Breyer MD, Moeckel GW, Harris RC (2007) Overexpression of cyclooxygenase-2 predisposes to podocyte injury. J Am Soc Nephrol 18(2):551–559

    Article  PubMed  CAS  Google Scholar 

  37. Xu L, Yang F, Lin R, Han C, Liu J, Ding Z (2014) Induction of M2 polarization in primary culture liver macrophages from rats with acute pancreatitis. PLoS ONE 9(9):e108014

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  38. Afiyani AA, Deemeh MR, Tavalaee M, Razi M, Bahadorani M, Shokrollahi B, Nasr-Esfahani MH (2014) Evaluation of heat-shock protein A2 (HSPA2) in male rats before and after varicocele induction. Mol Reprod Dev 81(8):766–776

    PubMed  CAS  Google Scholar 

  39. Razi M, Sadrkhanloo RA, Malekinejad H, Sarrafzadeh-Rezaei F (2012) Testicular biohistochemical alterations following experimental varicocele in rats. Iran J Reprod Med 10(3):209–218

    PubMed  PubMed Central  CAS  Google Scholar 

  40. Tremellen K (2008) Oxidative stress and male infertility—a clinical perspective. Hum Reprod Update 14(3):243–258

    Article  PubMed  CAS  Google Scholar 

  41. Khosravanian H, Razi M, Farokhi F, Khosravanian N (2015) Simultaneous administration of dexamethasone and vitamin E reversed experimental varicocele-induced impact in testicular tissue in rats; correlation with Hsp70-2 chaperone expression. Int Braz J Urol Off J Braz Soc Urol 41(4):773–790

    Article  Google Scholar 

  42. Kothari S, Thompson A, Agarwal A, du Plessis SS (2010) Free radicals: their beneficial and detrimental effects on sperm function. Indian J Exp Biol 48(5):425–435

    PubMed  CAS  Google Scholar 

  43. Razi M, Sadrkhanloo R-A, Malekinejad H, Sarafzadeh-Rezaei F (2011) Varicocele time-dependently affects DNA integrity of sperm cells: evidence for lower in vitro fertilization rate in varicocele-positive rats. Int J Fertil Steril 5(3):174–185

    PubMed  PubMed Central  Google Scholar 

  44. Wang X, Shen CL, Dyson MT, Eimerl S, Orly J, Hutson JC, Stocco DM (2005) Cyclooxygenase-2 regulation of the age-related decline in testosterone biosynthesis. Endocrinology 146(10):4202–4208

    Article  PubMed  CAS  Google Scholar 

  45. Turker Koksal I, Erdogru T, Gulkesen H, Sezer C, Usta M, Ciftcioglu A, Baykara M (2004) The potential role of inducible nitric oxide synthase (iNOS) activity in the testicular dysfunction associated with varicocele: an experimental study. Int Urol Nephrol 36(1):67–72

    Article  PubMed  Google Scholar 

  46. Costur P, Filiz S, Gonca S, Culha M, Gulecen T, Solakoglu S, Canberk Y, Caliskan E (2012) Expression of inducible nitric oxide synthase (iNOS) in the azoospermic human testis. Andrologia 44(Suppl 1):654–660

    Article  PubMed  CAS  Google Scholar 

  47. Lue Y, Wang C, Liu YX, Hikim AP, Zhang XS, Ng CM, Hu ZY, Li YC, Leung A, Swerdloff RS (2006) Transient testicular warming enhances the suppressive effect of testosterone on spermatogenesis in adult cynomolgus monkeys (Macaca fascicularis). J Clin Endocrinol Metab 91(2):539–545

    Article  PubMed  CAS  Google Scholar 

  48. Beltran B, Orsi A, Clementi E, Moncada S (2000) Oxidative stress and S-nitrosylation of proteins in cells. Br J Pharmacol 129(5):953–960

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  49. Guo J, Jia Y, Tao SX, Li YC, Zhang XS, Hu ZY, Chiang N, Lue YH, Hikim AP, Swerdloff RS, Wang C, Liu YX (2009) Expression of nitric oxide synthase during germ cell apoptosis in testis of cynomolgus monkey after testosterone and heat treatment. J Androl 30(2):190–199

    Article  PubMed  CAS  Google Scholar 

  50. Allen JD, Gow AJ (2009) Nitrite, NO and hypoxic vasodilation. Br J Pharmacol 158(7):1653–1654

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  51. Jourd’heuil D, Jourd’heuil FL, Kutchukian PS, Musah RA, Wink DA, Grisham MB (2001) Reaction of superoxide and nitric oxide with peroxynitrite. Implications for peroxynitrite-mediated oxidation reactions in vivo. J Biol Chem 276(31):28799–28805

    Article  PubMed  Google Scholar 

  52. Serhan CN, Savill J (2005) Resolution of inflammation: the beginning programs the end. Nat Immunol 6(12):1191–1197

    Article  PubMed  CAS  Google Scholar 

  53. Legler DF, Bruckner M, Uetz-von Allmen E, Krause P (2010) Prostaglandin E2 at new glance: novel insights in functional diversity offer therapeutic chances. Int J Biochem Cell Biol 42(2):198–201

    Article  PubMed  CAS  Google Scholar 

  54. Smyth EM, Grosser T, Wang M, Yu Y, FitzGerald GA (2009) Prostanoids in health and disease. J Lipid Res 50(Suppl):423–428

    Article  CAS  Google Scholar 

  55. Ricciotti E, FitzGerald GA (2011) Prostaglandins and inflammation. Arterioscler Thromb Vasc Biol 31(5):986–1000

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  56. Milla C, Yang S, Cornfield DN, Brennan ML, Hazen SL, Panoskaltsis-Mortari A, Blazar BR, Haddad IY (2004) Myeloperoxidase deficiency enhances inflammation after allogeneic marrow transplantation. Am J Physiol Lung Cell Mol Physiol 287(4):L706–L714

    Article  PubMed  CAS  Google Scholar 

  57. Lu Y, Wahl LM (2005) Oxidative stress augments the production of matrix metalloproteinase-1, cyclooxygenase-2, and prostaglandin E2 through enhancement of NF-kappa B activity in lipopolysaccharide-activated human primary monocytes. J Immunol 175(8):5423–5429

    Article  PubMed  CAS  Google Scholar 

  58. Luo C, Urgard E, Vooder T, Metspalu A (2011) The role of COX-2 and Nrf2/ARE in anti-inflammation and antioxidative stress: aging and anti-aging. Med Hypotheses 77(2):174–178

    Article  PubMed  CAS  Google Scholar 

  59. Paduch R, Kandefer-Szerszeń M (2011) Nitric oxide (NO) and cyclooxygenase-2 (COX-2) cross-talk in co-cultures of tumor spheroids with normal cells. Cancer Microenviron 4(2):187–198

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  60. Balderas E, Sánchez-Cárdenas C, Chávez JC, de la Vega Beltrán JL, Gómez-Lagunas F, Treviño CL, Darszon A (2013) The anti-inflammatory drug celecoxib inhibits T-type Ca2+ currents in spermatogenic cells yet it elicits the acrosome reaction in mature sperm. FEBS Lett 587(15):2412–2419

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgements

The authors wish to thank Department of Comparative Histology and Embryology for technical support and Faculty of Veterinary Medicine, Urmia University. Moreover, current manuscript is obtained from thesis no: 1408, which is proofed by Urmia University. Moreover, this research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.

Author information

Authors and Affiliations

  1. Division of Histology and Embryology, Department of Basic Science, Faculty of Veterinary Medicine, Urmia University, P.O. Box: 1177, Urmia, Iran

    Sara Mazhari, Mazdak Razi & Rajabali Sadrkhanlou

Authors
  1. Sara Mazhari
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Mazdak Razi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Rajabali Sadrkhanlou
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Mazdak Razi.

Ethics declarations

Conflict of interest

The authors declare that there are no conflicts of interest.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Mazhari, S., Razi, M. & Sadrkhanlou, R. Silymarin and celecoxib ameliorate experimental varicocele-induced pathogenesis: evidences for oxidative stress and inflammation inhibition. Int Urol Nephrol 50, 1039–1052 (2018). https://doi.org/10.1007/s11255-018-1862-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11255-018-1862-5

Keywords

Search

Navigation