Evaluation of the serum sex hormones levels and alkaline phosphatase activity in rats’ testis after administering of berberine in experimental varicocele

  • Hamed Najaran
  • Hassan Hassani BafraniEmail author
  • Hamid Rashtbari
  • Fatemeh Izadpanah
  • Mohammad Reza Rajabi
  • Hamed Haddad Kashani
  • Abouzar Mohammadi
Research Article


Current study was aimed to investigate the protective effect of berberine (BB) on the serum gonadotropin-releasing hormone (GnRH), follicle-stimulating hormone (FSH), luteinizing hormone (LH), inhibin B (INHB), testosterone (T) and alkaline phosphatase (Alk-p) activity in the testis of experimental varicocele-induced animals. For the current objective, 30 mature-male Wistar rats were randomly divided into control (n = 6 rats), control-sham (n = 6 rats) and experimental groups (n = 18 rats). The animals in the experimental groups were undergone experimental varicocele and simple laparotomy was conducted in control-sham group. 60 days after varicocele (VCL) induction the experimental group subdivided into: non-treated VCL-induced and 50 mg/kg and 100 mg/kg BB-treated groups (intra-peritoneally). Following 60 days, the animals were euthanized and serum levels of testosterone and testicular activity of alkaline phosphatase were measured. Non-treated VCL-induced animals indicated a significant (P < 0.05) reduction in serum levels of T and INHB and a remarkable (P < 0.05) increase in GnRH, FSH, LH and Alk-p activity compared to control and control-sham groups. Insignificant changes were found between control and control-sham groups. Meanwhile, each BB administered group showed a remarkable (P < 0.05) increase in serum levels of T and INHB and a significant (P < 0.05) decrease in GnRH, FSH, LH and alkaline phosphatase activity in testis tissue. According to the current findings, BB by increasing serum levels of testosterone and INHB increases the testicular endocrine capacity and protects Leydig cell against inflammatory and oxidant injury of varicocele. In addition, BB by inhibiting GnRH, FSH, LH and alkaline phosphatase activity, regulate the levels of serum sex hormones in experimental varicocele and reduces varicocele-induced inflammatory reactions.


Varicocele Berberine Gonadotropin-releasing hormone Inhibin B Testosterone Alkaline phosphatase 







Gonadotropin-releasing hormone


Follicle-stimulating hormone


Luteinizing hormone


Inhibin B




Alkaline phosphatase



We would like to appreciate Dr. Mazdak Razi, the staffs of Histology laboratory, for their kind technical support. Also the authors wish to thank Kashan University of medical sciences for financial supports.

Authors’ contributions

HN and HHB developed the concept and designed the study. HN was involved in subject recruitment and laboratory analysis. All other three authors were involved in data analysis and helped draft the manuscript. All authors read and approved the final manuscript.


The research grant provided by Research Deputy of Kashan University of Medical Sciences.

Compliance with ethical standards

Ethical statement

All procedures performed in studies involving animal participants were in accordance with the ethical standards of the institutional and national research committee and with the 1964 Helsinki declaration and its later amendments. The ethical reference number is kaums-94127.

Conflict of interest

This manuscript described has not been published before; not under consideration for publication anywhere else; and has been approved by all co-authors.

Availability of data and materials

The primary data for this study is available from the authors on direct request.


  1. Akkoyunlu G et al (2007) Immunolocalization of glial cell-derived neurotrophic factor (GDNF) and its receptor GFR-α1 in varicocele-induced rat testis. Acta Histochem 109(2):130–137Google Scholar
  2. Amini Mahabadi J et al (2013) Effect of diet contains sesame seed on adult Wistar rat testis. Int J Morphol 31(1):197–202Google Scholar
  3. Armagan A et al (2012) The effect of micronized purified flavonoid fraction on the prevention of testicular pathologies in adolescent rats with experimentally induced varicocele. J Urol 188(5):2007–2013Google Scholar
  4. Benoff S, Gilbert BR (2001) Varicocele and male infertility: part I. Hum Reprod Update 7(1):47–54Google Scholar
  5. Bhutada P et al (2011) Protection of cholinergic and antioxidant system contributes to the effect of berberine ameliorating memory dysfunction in rat model of streptozotocin-induced diabetes. Behav Brain Res 220(1):30–41Google Scholar
  6. Blevrakis E et al (2016) Impact of varicocele on biological markers of gonadal function. Hernia 20(3):435–439Google Scholar
  7. Cheng D et al (2006) Effects of epidermal growth factor on sperm content and motility of rats with surgically induced varicoceles. Asian J Androl 8(6):713–717Google Scholar
  8. Cheng F et al (2013) Berberine improves endothelial function by reducing endothelial microparticles-mediated oxidative stress in humans. Int J Cardiol 167(3):936–942Google Scholar
  9. Dun R et al (2015) Traditional Chinese herb combined with surgery versus surgery for varicocele infertility: a systematic review and meta-analysis. Evid-Based Complement Altern Med. Google Scholar
  10. Ferdosian M et al (2015) Identification of immunotopes against Mycobacterium leprae as immune targets using PhDTm-12mer phage display peptide library. Trop J Pharm Res 14(7):1153–1159Google Scholar
  11. Ghosh PK, York JP (1994) Changes in testicular testosterone and acid and alkaline phosphatase activity in testis and accessory sex organs after induction of varicocele in Noble rats. J Surg Res 56(3):271–276Google Scholar
  12. Goldstein M, Tanrikut C (2006) Microsurgical management of male infertility. Nat Rev Urol 3(7):381Google Scholar
  13. Gorelick JI, Goldstein M (1993) Loss of fertility in men with varicocele. Fertil Steril 59(3):613–616Google Scholar
  14. Guarino N, Tadini B, Bianchi M (2003) The adolescent varicocele: the crucial role of hormonal tests in selecting patients with testicular dysfunction. J Pediatr Surg 38(1):120–123Google Scholar
  15. Haddad-Kashani H et al (2012) Pharmacological properties of medicinal herbs by focus on secondary metabolites. Life Sci J 9(1):509–520Google Scholar
  16. Haddad Kashani H et al (2017) A novel chimeric endolysin with antibacterial activity against methicillin-resistant Staphylococcus aureus. Front Cell Infect Microbiol 7:290Google Scholar
  17. Hayes FJ et al (1998) Differential control of gonadotropin secretion in the human: endocrine role of inhibin. J Clin Endocrinol Metab 83(6):1835–1841Google Scholar
  18. Hosseini ES et al (2016) Purification of antibacterial CHAP K protein using a self-cleaving fusion tag and its activity against methicillin-resistant Staphylococcus aureus. Probiotics Antimicrob Proteins 8(4):202–210Google Scholar
  19. Hosseyni ES et al (2012) Mode of action of medicinal plants on diabetic disorders. Life Sci J 4(9):2776–2783Google Scholar
  20. Hsieh Y-S et al (2007) Protective effects of berberine against low-density lipoprotein (LDL) oxidation and oxidized LDL-induced cytotoxicity on endothelial cells. J Agric Food Chem 55(25):10437–10445Google Scholar
  21. Hudson RW, Crawford VA, Mckay DE (1981) The gonadotropin response of men with varicoceles to a four-hour infusion of gonadotropin-releasing hormone. Fertil Steril 36(5):633–637Google Scholar
  22. Jung HA et al (2009) Anti-Alzheimer and antioxidant activities of Coptidis Rhizoma alkaloids. Biol Pharm Bull 32(8):1433–1438Google Scholar
  23. Kashani HH, Moniri R (2015) Expression of recombinant pET22b-LysK-cysteine/histidine-dependent amidohydrolase/peptidase bacteriophage therapeutic protein in Escherichia coli BL21 (DE3). Osong Public Health Res Perspect 6(4):256–260Google Scholar
  24. Kashani HH et al (2013) Expression of galectin-3 as a testis inflammatory marker in vasectomised mice. Cell J (Yakhteh) 15(1):11Google Scholar
  25. Kashani HH et al (2018) Recombinant endolysins as potential therapeutics against antibiotic-resistant Staphylococcus aureus: current status of research and novel delivery strategies. Clin Microbiol Rev 31(1):e00071-17Google Scholar
  26. Kong W et al (2004) Berberine is a novel cholesterol-lowering drug working through a unique mechanism distinct from statins. Nat Med 10(12):1344Google Scholar
  27. Lotfi A et al (2016) Comparing the effects of two feeding methods on metabolic bone disease in newborns with very low birth weights. Glob J Health Sci 8(1):249Google Scholar
  28. Luo D-Y et al (2011) Effects of varicocele on testosterone, apoptosis and expression of StAR mRNA in rat Leydig cells. Asian J Androl 13(2):287Google Scholar
  29. Marmar JL (2001) Varicocele and male infertility: part II: the pathophysiology of varicoceles in the light of current molecular and genetic information. Hum Reprod Update 7(5):461–472Google Scholar
  30. Martin B, Jacoby F (1949) Diffusion phenomenon complicating the histochemical reaction for alkaline phosphatase. J Anat 83(Pt 4):351Google Scholar
  31. Miyaoka R, Esteves SC (2012) A critical appraisal on the role of varicocele in male infertility. Adv Urol 2012:597495Google Scholar
  32. Moghadam SE et al (2018) Neuroprotective effects of oxytocin hormone after an experimental stroke model and the possible role of calpain-1. J Stroke Cerebrovasc Dis 27(3):724–732Google Scholar
  33. Moghaddasi Mohammad S, Kashani HH, Azarbad Z (2012) Capparis spinosa L. propagation and medicinal uses. Life Sci J 9(4):684–686Google Scholar
  34. Moshtaghion S-M et al (2013) Silymarin protects from varicocele-induced damages in testis and improves sperm quality: evidence for E2f1 involvement. Syst Biol Reprod Med 59(5):270–280Google Scholar
  35. Naeini ZK, Bafrani HH, Nikzad H (2014) Evaluation of ebselen supplementation on cryopreservation medium in human semen. Iran J Reprod Med 12(4):249Google Scholar
  36. Nikzad H et al (2013) Expression of galectin-8 on human endometrium: molecular and cellular aspects. Iran J Reprod Med 11(1):65Google Scholar
  37. O’donnell L et al (1994) Testosterone promotes the conversion of round spermatids between stages VII and VIII of the rat spermatogenic cycle. Endocrinology 135(6):2608–2614Google Scholar
  38. Pierik FH et al (2001) Increased serum inhibin B levels after varicocele treatment. Clin Endocrinol 54(6):775–780Google Scholar
  39. Plant TM, Marshall GR (2001) The functional significance of FSH in spermatogenesis and the control of its secretion in male primates. Endocr Rev 22(6):764–786Google Scholar
  40. Plymate SR, Paulsen CA, McLachlan R (1992) Relationship of serum inhibin levels to serum follicle stimulating hormone and sperm production in normal men and men with varicoceles. J Clin Endocrinol Metab 74(4):859–864Google Scholar
  41. Račková L et al (2004) Antiradical and antioxidant activities of alkaloids isolated from Mahonia aquifolium. Structural aspects. Bioorg Med Chem 12(17):4709–4715Google Scholar
  42. Rajfer J et al (1987) Inhibition of testicular testosterone biosynthesis following experimental varicocele in rats. Biol Reprod 36(4):933–937Google Scholar
  43. Razi M et al (2011) Histological impact of long term varicocele-induction on right and left testes in rat (evidence for the reduction of sperm quality and mating abilities). In: Veterinary research forum. Faculty of Veterinary Medicine, Urmia UniversityGoogle Scholar
  44. Rege N et al (1979) Serum gonadotropins and testosterone in infertile patients with varicocele. Fertil Steril 31(4):413–416Google Scholar
  45. Ricci J-E et al (2004) Disruption of mitochondrial function during apoptosis is mediated by caspase cleavage of the p75 subunit of complex I of the electron transport chain. Cell 117(6):773–786Google Scholar
  46. Romeo C et al (2007) Altered serum inhibin b levels in adolescents with varicocele. J Pediatr Surg 42(2):390–394Google Scholar
  47. Said TM et al (2012) Oxidative stress, DNA damage, and apoptosis in male infertility. In: Agarwal A, Aitken RJ, Alvarez JG (eds) Studies on men’s health and fertility. Humana Press Inc., NJ, pp 433–448Google Scholar
  48. Shan L et al (1995) Effects of luteinizing hormone (LH) and androgen on steady state levels of messenger ribonucleic acid for LH receptors, androgen receptors, and steroidogenic enzymes in rat Leydig cell progenitors in vivo. Endocrinology 136(4):1686–1693Google Scholar
  49. Sharif MR et al (2016) The effect of a yeast probiotic on acute diarrhea in children. Probiotics Antimicrob Proteins 8(4):211–214Google Scholar
  50. Sharif A et al (2017) The role of probiotics in the treatment of dysentery: a randomized double-blind clinical trial. Probiotics Antimicrob Proteins 9(4):380–385Google Scholar
  51. Shirwaikar A et al (2006) In vitro antioxidant studies on the benzyl tetra isoquinoline alkaloid berberine. Biol Pharm Bull 29(9):1906–1910Google Scholar
  52. Sirvent J et al (1990) Leydig cell in idiopathic varicocele. Eur Urol 17:257–261Google Scholar
  53. Sofikitis N et al (1992) Surgical repair versus medical treatment of varicocele in the rat: pharmacological manipulation of the varicocelized testicle. Eur Urol 22:44–52Google Scholar
  54. Sokol RZ (2009) Endocrinology of male infertility: evaluation and treatment. Semin Reprod Med 27(2):149–158Google Scholar
  55. Tejada RI et al (1984) A test for the practical evaluation of male fertility by acridine orange (AO) fluorescence. Fertil Steril 42(1):87–91Google Scholar
  56. Warner JN, Frey KA (2013) The well-man visit: addressing a man’s health to optimize pregnancy outcomes. J Am Board Fam Med 26(2):196–202Google Scholar
  57. Weinbauer GF, Nieschlag E (1995) Gonadotropin control of testicular germ cell development. Adv Exp Med Biol 317:55–65Google Scholar
  58. Zhang J et al (2011) Oxidative stress: role in acetamiprid-induced impairment of the male mice reproductive system. Agric Sci China 10(5):786–796Google Scholar
  59. Zhang L et al (2016) Effects of Morinda officinalis polysaccharide on experimental varicocele rats. Evid-Based Complement Altern Med. Google Scholar
  60. Zhang J et al (2018) Roles of Fas/FasL-mediated apoptosis and inhibin B in the testicular dysfunction of rats with left-side varicocele. Andrologia 50(2):e12850Google Scholar

Copyright information

© Institute of Korean Medicine, Kyung Hee University 2019

Authors and Affiliations

  • Hamed Najaran
    • 1
  • Hassan Hassani Bafrani
    • 1
    • 2
    Email author
  • Hamid Rashtbari
    • 1
  • Fatemeh Izadpanah
    • 3
  • Mohammad Reza Rajabi
    • 4
  • Hamed Haddad Kashani
    • 2
  • Abouzar Mohammadi
    • 5
  1. 1.Gametogenesis Research CenterKashan University of Medical SciencesKashanIran
  2. 2.Anatomical Sciences Research CenterKashan University of Medical SciencesKashanIran
  3. 3.Food and Drug Laboratory Research Center and Food and Drug Reference Control Laboratories CenterFood and Drug Administration of Iran, MOH & METehranIran
  4. 4.Faculty of MedicineShahed University of Medical SciencesTehranIran
  5. 5.Surgical Technology Department, Nursing and Midwifery CollegeKashan University of Medical SciencesKashanIran

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