Troxerutin affects the male fertility in prepubertal type 1 diabetic male rats

Document Type: Original Article


1 Department of Physiology, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran

2 Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran

3 Department of Basic Sciences, Faculty of Veterinary Medicine, University of Tabriz, Tabriz, Iran

4 Department of physiology, Faculty of medicine, Tabriz University of Medical Sciences, Tabriz, Iran

5 Tuberculosis and Lung Diseases Research Center, Tabriz University of Medical Sciences, Tabriz, Iran


Objective(s): Diabetes can gradually cause damage to the function and structure of male gonads. This survey was conducted to investigate the effect of troxerutin on hormonal changes, serum oxidative stress indices, and testicular function and structure in prepubertal diabetic rats.
Materials and Methods: Fifty prepubertal (6 weeks old) male Wistar rats were divided into five groups including Control, Troxerutin, Diabetic, Diabetic+Troxerutin, and Diabetic+Insulin. Type I diabetes was induced by 55 mg/kg of streptozotocin intraperitoneally. The groups were treated with 150 mg/kg/day troxerutin via oral gavage or 4-6 IU/day insulin via subcutaneous injection for 4 consecutive weeks. Blood sugar (BS) and serum levels of insulin, FSH, LH, testosterone, glutathione peroxidase (GPX), superoxide dismutase (SOD), malondialdehyde (MDA), and total antioxidant capacity (TAC) were analyzed. Testis and epididymis were removed for histopathologic study and analysis of sperm parameters.
Results: Troxerutin significantly reduced the BS in the diabetic group similar to insulin but could not affect insulin, FSH, or LH significantly. Troxerutin caused a significant increase in testosterone and GPX but had no significant effect on serum MDA, TAC, and SOD levels. In addition, troxerutin had a better effect than insulin on diabetes-induced testicular structural damage. Sperm analysis results also revealed that troxerutin and insulin could improve sperm number, motility, and viability in diabetic rats.
Conclusion: According to these results, it can be derived that administration of troxerutin is a suitable protective strategy for side effects of diabetes in testis of prepubertal diabetic male rats.


Main Subjects

1. Baluchnejadmojarad T, Jamali-Raeufy N, Zabihnejad S, Rabiee N, Roghani M. Troxerutin exerts neuroprotection in 6-hydroxydopamine lesion rat model of Parkinson’s disease: Possible involvement of PI3K/ERβ signaling. Eur J Pharmacol 2017; 801:72-78.
2. Ding GL, Liu Y, Liu ME, Pan JX, Guo MX, Sheng JZ, Huang HF. The effects of diabetes on male fertility and epigenetic regulation during spermatogenesis. Asian J Androl 2015; 17:948-53.
3. Kolahian S, Sadri H, Larijani A, Hamidian G, Davasaz A. Supplementation of diabetic rats with leucine, zinc, and chromium: effects on function and histological structure of testes. Int J Vitam Nutr Res 2015; 85:311-21.
4. Butler AE, Janson J, Bonner-Weir S, Ritzel R, Rizza RA, Butler PC. β-Cell deficit and increased β-cell apoptosis in humans with type 2 diabetes. Diabetes 2003; 52:102-110.
5. Thomas MC, Cooper ME, Zimmet P. Changing epidemiology of type 2 diabetes mellitus and associated chronic kidney disease. Nat Rev Nephrol 2016; 12:73-81.
6. Ashcroft FM, Rorsman P. Diabetes mellitus and the β cell: the last ten years. Cell 2012; 148:1160-1171.
7. Regnell SE, Lernmark A. Early prediction of autoimmune (type 1) diabetes. Diabetologia 2017; 60:1370-1381.
8. Zurita-Cruz JN, Nishimura-Meguro E, Villasís-Keever MA, Hernández-Méndez ME, Garrido-Magaña E, Rivera-Hernández AJ. Influence of the informal primary caretaker on glycemic control among prepubertal pediatric patients with type 1 diabetes mellitus. J Pediatr (Rio J) 2017; 93:136-141.
9. Sexton WJ, Jarow JP. Effect of diabetes mellitus upon male reproductive function. Urology 1997; 49:508-513.
10. Ali ST, Shaikh RN, Ashfaqsiddiqi N, Siddiqi PQ. Serum and urinary levels of pituitary-gonadal hormones in insulin-dependent and non-insulin-dependent diabetic males with and without neuropathy. Arch Androl 1993; 30:117-123.
11. Ramalho-Santos J, Amaral S, Oliveira PJ. Diabetes and the impairment of reproductive function: possible role of mitochondria and reactive oxygen species. Curr Diabetes Rev 2008; 4:46-54.
12. Giacco F, Brownlee M. Oxidative stress and diabetic complications. Circ Res 2010; 107:1058-1070.
13. Tiwari BK, Pandey KB, Abidi AB, Rizvi SI. Markers of oxidative stress during diabetes mellitus. J Biomark 2013; 2013:1-8.
14. Watcho P, Mbiakop UC, Jeugo HG, Wankeu M, Nguelefack TB, Carro-Juarez M, Kamanyi A. Delay of ejaculation induced by Bersama engleriana in nicotinamide/streptozotocin-induced type 2 diabetic rats. Asian Pac J Trop Med 2014; 7:S603-S609.
15. Ryan JG, Gajraj J. Erectile dysfunction and its association with metabolic syndrome and endothelial function among patients with type 2 diabetes mellitus. J Diabetes Complications 2012; 26:141-147.
16. Abd El-Twab SM, Mohamed HM, Mahmoud AM. Taurine and pioglitazone attenuate diabetes-induced testicular damage by abrogation of oxidative stress and up-regulation of the pituitary–gonadal axis. Can J Physiol Pharmacol 2016; 94:651-661.
17. Pomjunya A, Ratthanophart J, Fungfuang W. Effects of Vernonia cinerea on reproductive performance in streptozotocin-induced diabetic rats. J Vet Med Sci 2017; 79:572-578.
18. Wankeu-Nya M, Florea A, Bâlici S, Watcho P, Matei H, Kamanyi A. Dracaena arborea alleviates ultra-structural spermatogenic alterations in streptozotocin-induced diabetic rats. BMC Complement Altern Med 2013; 13:71-79.
19. De A, Singh MF, Singh V, Ram V, Bisht S. Treatment effect of l-Norvaline on the sexual performance of male rats with streptozotocin induced diabetes. Eur J Pharmacol 2016; 771:247-254.
20. Rovira-Llopis S, Bañuls C, de Marañon AM, Diaz-Morales N, Jover A, Garzon S, Rocha M, Victor VM, Hernandez-Mijares A. Low testosterone levels are related to oxidative stress, mitochondrial dysfunction and altered subclinical atherosclerotic markers in type 2 diabetic male patients. Free Radic Biol Med 2017; 108:155-162.
21. Patel DK, Kumar R, Laloo D, Hemalatha S. Diabetes mellitus: an overview on its pharmacological aspects and reported medicinal plants having antidiabetic activity. Asian Pac J Trop Biomed 2012; 2:411-420.
22. Goyal M. Traditional plants used for the treatment of diabetes mellitus in Sursagar constituency, Jodhpur, Rajasthan–An ethnomedicinal survey. J Ethnopharmacol 2015; 174:364-368.
23. Fan SH, Zhang ZF, Zheng YL, Lu J, Wu DM, Shan Q, Hu B, Wang YY. Troxerutin protects the mouse kidney from d-galactose-caused injury through anti-inflammation and anti-oxidation. Int Immunopharmacol 2009; 9:91-96.
24. Badalzadeh R, Layeghzadeh N, Alihemmati A, Mohammadi M. Beneficial effect of troxerutin on diabetes-induced vascular damages in rat aorta: histopathological alterations and antioxidation mechanism. Int J Endocrinol Metab 2015; 13: e25969.
25. Panat NA, Maurya DK, Ghaskadbi SS, Sandur SK. Troxerutin, a plant flavonoid, protects cells against oxidative stress-induced cell death through radical scavenging mechanism. Food Chem 2016; 194:32-45.
26. Lu J, Wu DM, Zheng YL, Hu B, Cheng W, Zhang ZF, Li MQ. Troxerutin counteracts domoic acid–induced memory deficits in mice by inhibiting CCAAT/enhancer binding protein β–mediated inflammatory response and oxidative stress. J Immunol 2013; 190:3466-3479.
27. Zamanian M, Hajizadeh MR, Esmaeili Nadimi A, Shamsizadeh A, Allahtavakoli M. Antifatigue effects of troxerutin on exercise endurance capacity, oxidative stress and matrix metalloproteinase‐9 levels in trained male rats. Fund Clin Pharmacol 2017; 31:447-455.
28. Liu C-M, Ma J-Q, Lou Y. Chronic administration of troxerutin protects mouse kidney against D-galactose-induced oxidative DNA damage. Food Chem Toxicol 2010; 48:2809-2817.
29. Yu Y, Zheng G. Troxerutin protects against diabetic cardiomyopathy through NF‑κB/AKT/IRS1 in a rat model of type 2 diabetes. Mol Med Rep 2017; 15:3473-3478.
30. Zhang ZF, Fan SH, Zheng YL, Lu J, Wu DM, Shan Q, Hu B. Troxerutin protects the mouse liver against oxidative stress-mediated injury induced by D-galactose. J Agric Food Chem 2009; 57:7731-7736.
31. Gohel MS, Davies AH. Pharmacological agents in the treatment of venous disease: an update of the available evidence. Curr Vasc Pharmacol 2009; 7:303-308.
32. Elangovan P, Jalaludeen AM, Ramakrishnan R, Pari L. Protective effect of troxerutin on nickel-induced testicular toxicity in wistar rats. J Environ Pathol Toxicol Oncol 2016; 35.133-46.
33. Kaya H, Sezik M, Ozkaya O, Dittrich R, Siebzehnrubl E, Wildt L. Lipid peroxidation at various estradiol concentrations in human circulation during ovarian stimulation with exogenous gonadotropins. Horm Metab Res 2004; 36:693-695.
34. Choi WS, Kwon OS, Cho SY, Paick JS, Kim SW. Effect of chronic administration of PDE5 combined with glycemic control on erectile function in streptozotocin‐induced diabetic Rats. J Sex Med 2015; 12:600-610.
35. Alipour MR, Khamaneh AM, Yousefzadeh N, Mohammad-nejad D, Soufi FG. Upregulation of microRNA-146a was not accompanied by downregulation of pro-inflammatory markers in diabetic kidney. Mol Biol Rep 2013; 40:6477-6483.
36. Amaral S, Moreno AJ, Santos MS, Seiça R, Ramalho-Santos J. Effects of hyperglycemia on sperm and testicular cells of Goto-Kakizaki and streptozotocin-treated rat models for diabetes. Theriogenology 2006; 66:2056-2067.
37. Oger P, Yazbeck C, Gervais A, Dorphin B, Gout C, Jacquesson L, Ayel JP, Kahn V, Rougier N. Adverse effects of hepatitis B virus on sperm motility and fertilization ability during IVF. Reprod Biomed Online 2011; 23:207-212.
38. Keegan BR, Barton S, Sanchez X, Berkeley AS, Krey LC, Grifo J. Isolated teratozoospermia does not affect in vitro fertilization outcome and is not an indication for intracytoplasmic sperm injection. Fertil Steril 2007; 88:1583-1588.
39. Bahmanzadeh M, Abolhassani F, Amidi F, Ejtemaiemehr Sh, Salehi M, Abbasi M. The effects of nitric oxide synthase inhibitor (L-NAME) on epididymal sperm count, motility, and morphology in varicocelized rat. Daru 2008; 16:23-28.
40. Organization WH, WHO laboratory manual for the examination and processing of human semen 2010.
41. Johnsen SG. Testicular biopsy score count–a method for registration of spermatogenesis in human testes: normal values and results in 335 hypogonadal males. Horm Res Paediatr 1970; 1:2-25.
42. Oghbaei H, Alipour MR, Hamidian G, Ahmadi M, Ghorbanzadeh V, Keyhanmanesh R. Two months sodium nitrate supplementation alleviates testicular injury in streptozotocin-induced diabetic male rats. Exp Physiol 2018; doi: 10.1113/EP087198.
43. Keyhanmanesh R, Hamidian G, Alipour MR, Ranjbar M, Oghbaei H. Protective effects     of sodium nitrate against testicular apoptosis and spermatogenesis impairments in streptozotocin-induced diabetic male rats. Life Sci 2018; 211:63-73.
44. Samir Zahkok, Nehal Abo-Elnaga, Amel FM Ismail, Esraa Mousa. Studies on fertility of diabetic male rats treated with olive leaves extract. J Biomed Pharmaceut Res 2016; 5:18-27.
45. Ahmed, N., Advanced glycation endproducts—role in pathology of diabetic complications. Diabetes Res Clin Pract 2005; 67:3-21.
46. Jakuš, V. and N. Rietbrock, Advanced glycation end-products and the progress of diabetic vascular complications. Physiol Res 2004; 53:131-142.
47. Tremellen K. Oxidative stress and male infertility—a clinical perspective. Human Reproduction Update 2008; 14:243-258.
48. Maedler K, Carr RD, Bosco D, Zuellig RA, Berney T, Donath MY. Sulfonylurea induced β-cell apoptosis in cultured human islets. J Clin Endocrin Metab 2005; 90:501-506.
49. Adaramoye O, Akanni O, Adesanoye O, Labo-Popoola O, Olaremi O. Evaluation of toxic effects of metformin hydrochloride and glibenclamide on some organs of male rats. Niger J Physiol Sci 2012; 27:137-144.
50. Alves MG, Socorro S, Silva J, Barros A, Sousa M, Cavaco JE, Oliveira PF. In vitro cultured human Sertoli cells secrete high amounts of acetate that is stimulated by 17β-estradiol and suppressed by insulin deprivation. Biochim Biophys Acta 2012; 1823:1389-1394.
51. Oliveira PF, Alves MG, Rato L, Laurentino S, Silva J, Sá R, Barros A, Sousa M, Carvalho RA, Cavaco JE, Socorro S. Effect of insulin deprivation on metabolism and metabolism-associated gene transcript levels of in vitro cultured human Sertoli cells. Biochim Biophys Acta 2012; 1820:84-89.
52. Nakayama Y, Yamamoto T, Abé SI. IGF-I, IGF-II and insulin promote differentiation of spermatogonia to primary spermatocytes in organ culture of newt testes. Int J Dev Biol 1999; 43:343-347.
53. Geetha R, Yogalakshmi B, Sreeja S, Bhavani K, Anuradha CV. Troxerutin suppresses lipid abnormalities in the heart of high-fat–high-fructose diet-fed mice. Mol cell biochem 2014; 387:123-134.
54. Lu J, Wu DM, Zheng ZH, Zheng YL, Hu B, Zhang ZF. Troxerutin protects against high cholesterol-induced cognitive deficits in mice. Brain 2011; 134:783-797.
55. Kawamura N, Ookawara T, Suzuki K, Konishi K, Mino M, Taniguchi N. Increased glycated Cu, Zn-superoxide dismutase levels in erythrocytes of patients with insulin-dependent diabetis mellitus. J Clin Endocrin Metab 1992; 74:1352-1354.
56. Karimi J, Goodarzi MT, Tavilani H, Khodadadi I, Amiri I. Relationship between advanced glycation end products and increased lipid peroxidation in semen of diabetic men. Diabetes Res Clin Pract 2011; 91: 61-66.
57. Ballester J, Muñoz MC, Domínguez J, Rigau T, Guinovart JJ, Rodríguez-Gil JE. Insulin‐dependent diabetes affects testicular function by FSH‐and LH‐linked mechanisms. J Androl 2004; 25: 706-719.
58. La Vignera S, Condorelli R, Vicari E, D’Agata R, Calogero AE. Diabetes mellitus and sperm parameters. J Androl 2012; 33:145-153.
59. Kim ST, Moley KH. Paternal effect on embryo quality in diabetic mice is related to poor sperm quality and associated with decreased glucose transporter expression. Reproduction 2008; 136:313-322.
60. Baccetti B, La Marca A, Piomboni P, Capitani S, Bruni E, Petraglia F, De Leo V. Insulin-dependent diabetes in men is associated with hypothalamo-pituitary derangement and with impairment in semen quality. Hum Reprod 2002; 17:2673-2677.
61. Vinothkumar R, Vinoth Kumar R, Sudha M, Viswanathan P, Balasubramanian T, Nalini N. Modulatory effect of troxerutin on biotransforming enzymes and preneoplasic lesions induced by 1, 2-dimethylhydrazine in rat colon carcinogenesis. Exp Mol Pathol 2014; 96: 15-26.
62. Kanter M, Aktas C, Erboga M. Protective effects of quercetin against apoptosis and oxidative stress in streptozotocin-induced diabetic rat testis. Food Chem Toxicol 2012; 50:719-725.
63. Lu J, Wu DM, Hu B, Cheng W, Zheng YL, Zhang ZF, Ye Q, Fan SH, Shan Q, Wang YJ. Chronic administration of troxerutin protects mouse brain against D-galactose-induced impairment of cholinergic system. Neurobiol Learn Mem 2010; 93:157-164.