Preventive effects of quercetin against inflammation and apoptosis in cyclophosphamide-induced testicular damage

Document Type : Original Article

Authors

Department of Histology and Embryology, Faculty of Medicine, Trakya University, Edirne, Turkey

Abstract

Objective(s): We aimed to investigate the effects of quercetin (QRC) against cyclophosphamide (CP)-induced testicular damage and how it interacts with apoptotic and inflammatory signaling pathways.
Materials and Methods: Forty male Wistar rats were randomly divided into four groups, 10 in each group; Control group (corn oil, intragastrically, 14 days), QRC group (100 mg/kg QRC, dissolved in corn oil, 14 days), CP group (200 mg/kg CP, intraperitoneally, single dose on the 7th day), and CP+QRC group (100 mg/kg QRC, intragastrically, 14 days and 200 mg/kg CP, intraperitoneally, single dose on the 7th day). Animals were sacrificed one day after the last QRC application and the effects of quercetin were evaluated by histological, morphometrical, and hormonal parameters. Also, nuclear factor kappa B (NFkB), nuclear factor erythroid 2 related factor 2 (Nrf2), Bcl-2 associated X protein (Bax), and B-cell lymphoma-2 (Bcl-2) immunoreactivities were evaluated immunohistochemically.
Results: CP increased the testicular weight/body weight ratio, significantly decreasing body weights and testicular weights. All hormone levels were also reduced significantly. Morphometrically, seminiferous tubules diameter and germinal epithelial thickness decreased, while a significant increase was determined in interstitial field width in addition to histological damage. Furthermore, immunohistochemical findings also indicated that NFkB and Bax immunoreactivity were increased in the CP group, whereas significant decrease was seen in Nrf2 and Bcl-2 immunoreactivity. Apoptotic cell and tubule index were reduced in CP. QRC ensured improvement in all findings.
Conclusion: Data showed us, that QRC may have preventive effects in CP-induced testicular damage by acting on NFkB, Nrf2,  Bax, and Bcl-2 pathways.

Keywords

Main Subjects


1. Zhao H, Jin B, Zhang X, Cui Y, Sun D, Gao C et al. Yangjing capsule ameliorates spermatogenesis in male mice exposed to cyclophosphamide. Evid Based Complement Alternat Med 2015; 980583:1-8.
2. Abd El Tawab AM, Shahin NN, Abdel Mohsen MM. Protective effect of Satureja montana extract on cyclophosphamide-induced testicular injury in rats. Chem Biol Interact 2014; 224:196-205.
3. Şekeroğlu V, Aydın B, Şekeroğlu ZA. Viscum album L. extract and quercetin reduce cyclophosphamide-induced cardiotoxicity, urotoxicity and genotoxicity in mice. Asian Pacific J Cancer Prev 2011; 12:2925-2931.
4. Ebokaiwe AP, Obasi DO, Njoku RC, Osawe S. Cyclophosphamide-induced testicular oxidative-inflammatory injury is accompanied by altered immunosuppressive indoleamine 2, 3-dioxygenase in wister rats: influence of dietary quercetin. Andrologia 2022; 54:e14341.
5. Rezaei S, Hosseinimehr SJ, Zargari M, Karimpour Malekshah A, Mirzaei M, Talebpour Amiri F. Protective effects of sinapic acid against cyclophosphamide-induced testicular toxicity via inhibiting oxidative stress, caspase-3 and NF-kB activity in BALB/c mice. Andrologia 2021; 53:1-9.
6. Blumenfeld Z, von Wolff M. GnRH-analogues and oral contraceptives for fertility preservation in women during chemotherapy. Hum Reprod Update 2008; 14:543-552. 
7. Adana MY, Imam A, Bello AA, Sunmonu OE, Alege EP, Onigbolabi OG, et al. Oral thymoquinone modulates cyclophosphamide-induced testicular toxicity in adolescent Wistar rats. Andrologia 2022; 54:e14368.
8. Cao Y, Wang X, Li S, Wang H, Yu L, Wang P. The effects of L-carnitine against cyclophosphamide-induced injuries in mouse testis. Basic Clin Pharmacol Toxicol 2017; 120:152-158.
9. Jalali AS, Hasanzadeh S, Malekinejad H. Crataegus monogyna aqueous extract ameliorates cyclophosphamide-induced toxicity in rat testis: stereological evidences. Acta Med Iran 2012; 50:1-8.
10. Lu WP, Mei XT, Wang Y, Zheng YP, Xue YF, Xu DH. Zn(II) curcumin protects against oxidative stress, deleterious changes in sperm parameters and histological alterations in a male mouse model of cyclophosphamide-induced reproductive damage. Environ Toxicol Pharmacol 2015; 39:515-524.
11. Kaya C, Barbaros Baseskioglu A, Yigitaslan S, Yasemin Ozatik F, Ozatik O, Uslu S. The therapeutic potential of amifostine on cyclophosphamide-induced testicular dysfunction in rats: an experimental study. Int J Reprod Biomed 2019; 17:245-252.
12. Parandin R, Ghowsi M, Dadbod A. Protective effects of hydroalcoholic extract of Rosa canina L. fruit on cyclophosphamide-induced testicular toxicity in mice. Avicenna J Phytomed 2023; 13:7-17.
13. Liu F, Li XL, Lin T, He DW, Wei GH, Liu JH et al. The cyclophosphamide metabolite, acrolein, induces cytoskeletal changes and oxidative stress in sertoli cells. Mol Biol Rep 2012; 39:493-500.
14. Lawrence T. The nuclear factor NF-kappaB pathway in inflammation. Cold Spring Harb Perspect Biol 2009; 1:1-10.
15. Tu W, Wang H, Li S, Liu Q, Sha H. The anti-inflammatory and anti-oxidant mechanisms of the keap1/Nrf2/ARE signaling pathway in chronic diseases. Aging Dis 2019; 10:637-651.
16. Maremanda KP, Khan S, Jena G. Zinc protects cyclophosphamide-induced testicular damage in rat: involvement of metallothionein, tesmin and Nrf2. Biochem Biophys Res Commun 2014; 445:591-596.
17. Li W, Khor TO, Xu C, Shen G, Jeong WS, Yu S, et al. Activation of Nrf2-antioxidant signaling attenuates NFkappaB-inflammatory response and elicits apoptosis. Biochem Pharmacol 2008; 76:1485-1489.
18. Koohsari M, Ahangar N, Mohammadi E, Talebpour Amiri F, Shaki F. Effects of tramadol administration on male reproductive toxicity in wistar rats the role of oxidative stress, mitochondrial dysfunction, apoptosis-related gene expression, and nuclear factor kappa B signalling. Bratisl Lek Listy 2020; 121:400-410.
19. Yuan D, Wang H, He H, Jia L, He Y, Wang T, et al. Protective effects of total flavonoids from epimedium on the male mouse reproductive system against cyclophosphamide-induced oxidative injury by up-regulating the expressions of SOD3 and GPX1. Phytother Res 2014; 28:88-97.
20. Liu X, Song L. Quercetin protects human liver cells from o,p›-DDT-induced toxicity by suppressing Nrf2 and NADPH oxidase-regulated ROS production. Food Chem Toxicol 2022; 161: 112849.
21. Onur M, Yalçın E, Çavuşoğlu K, Acar A. Elucidating the toxicity mechanism of AFM2 and the protective role of quercetin in albino mice. Sci Rep 2023; 13:1237-1253.
22. Alizadeh SR, Ebrahimzadeh MA. Quercetin derivatives: drug design, development, and biological activities, a review. Eur J Med Chem 2022; 229: :114068.
23. Xu D, Hu MJ, Wang YQ, Cui YL. Antioxidant activities of quercetin and its complexes for medical application. Molecules 2019; 24:1-15.
24. Hu J, Yu Q, Zhao F, Ji J, Jiang Z, Chen X et al. Protection of quercetin against triptolide-induced apoptosis by suppressing oxidative stress in rat leydig cells. Chemi Biol Interact 2015; 240:38-46.
25. Şengül E, Gelen V, Gedikli S, Özkanlar S, Gür C, Çelebi F et al. The protective effect of quercetin on cyclophosphamide-Induced lung toxicity in rats. Biomed Pharmacother 2017; 92:303-307.
26. Nouri HS, Azarmi Y, Movahedin M. Effect of growth hormone on testicular dysfunction induced by methotrexate in rats. Andrologia 2009; 41:105–110.
27. Delen O, Uz YH. Protective effect of pyrrolidine dithiocarbamate against methotrexate-induced testicular damage. Hum Exp Toxicol 2021; 40:164-177.
28. Orazizadeh M, Khorsandi L, Absalan F, Hashemitabar M, Daneshi E. Effect of beta-carotene on titanium oxide nanoparticles-induced testicular toxicity in mice. J Assist Reprod Genet 2014; 31:561–568.
29. Uz YH, Murk W, Yetkin CE, Kayisli UA, Arici A. Expression and role of interleukin-23 in human endometrium throughout the menstrual cycle and early pregnancy. J Reprod Immunol 2010; 87:21-27.
30. Uzun-Goren D, Uz YH. Protective effect of curcumin against gentamicin-induced nephrotoxicity mediated by p38 MAPK, nuclear factor- kappa B, nuclear factor erythroid 2-related factor 2. Iran J Kidney Dis 2022; 16:96-107.
31. Yazdani I, Majdani R, Ghasemnejad-Berenji M, Dehpour AR. Comparison of multiple doses of cyclosporine A on germ cell apoptosis and epididymal sperm parameters after testicular ischemia/reperfusion in rats. Exp Mol Pathol 2019; 110:1-8.
32. Umamaheswari S, Girish C, Basu D. Effects of cleistanthus collinus on the reproductive system of male wistar rats. JBRA Assist Reprod 2022; 26:460-468.
33. Oyagbemi AA, Omobowale TO, Saba AB, Adedara IA, Olowu ER, Akinrinde AS et al. Gallic acid protects against cyclophosphamide-induced toxicity in testis and epididymis of rats. Andrologia 2016; 48:393-401.
34. Can S, Çetik Yıldız S, Keskin C, Şahintürk V, Cengiz M, Appak Başköy S, et al. Investigation into the protective effects of Hypericum triquetrifolium turra seed against cyclophosphamide-induced testicular injury in sprague dawley rats. Drug Chem Toxicol 2022; 45:1679-1686.
35. Ebokaiwe AP, Ushang OR, Ogunwa TH, Kikiowo B, Olusanya O. Quercetin attenuates cyclophosphamide induced-immunosuppressive indoleamine 2,3-dioxygenase in the hippocampus and cerebral cortex of male wister rats. J Biochem Mol Toxicol 2022; 36: :e23179.
36. Motawi TM, Sadik NA, Refaat A. Cytoprotective effects of DL-alpha-lipoic acid or squalene on cyclophosphamide-induced oxidative injury: an experimental study on rat myocardium, testicles and urinary bladder. Food Chem Toxicol 2010; 48:2326-2336.
37. Kim SH, Lee IC, Ko JW, Shin IS, Moon C, Kim SH et al. Mechanism of protection by diallyl disulfide against cyclophosphamide-induced spermatotoxicity and oxidative stress in rats. Mol Cell Toxicol 2016; 12:301-312.
38. Ekeleme-Egedigwe CA, Famurewa AC, David EE, Eleazu CO, Egedigwe UO. Antioxidant potential of garlic oil supplementation prevents cyclophosphamide-induced oxidative testicular damage and endocrine depletion in rats. J Nutr Intermed Metab 2019; 18:1-6.
39. Hamzeh M, Hosseinimehr SJ, Karimpour A, Mohammadi HR, Khalatbary AR, Talebpour Amiri F. Cerium oxide nanoparticles protect cyclophosphamide-induced testicular toxicity in mice. Int J Prev Med 2019; 10:1-9.
40. Anan HH, Zidan RA, Abd El-Baset SA, Ali MM. Ameliorative effect of zinc oxide nanoparticles on cyclophosphamide induced testicular injury in adult rat. Tissue Cell 2018; 54:80-93.
41. Al-Omair MA, Sedky A, Ali A, Elsawy H. Ameliorative potentials of quercetin against lead-induced hematological and testicular alterations in albino rats. Chin J Physiol 2017; 60:54-61.
42. Mohammadi F, Nikzad H, Taghizadeh M, Taherian A, Azami-Tameh A, Hosseini SM et al. Protective effect of zingiber officinale extract on rat testis after cyclophosphamide treatment. Andrologia 2014; 46:680-686.
43. Hosseini A, Zare S, Borzouei Z, Ghaderi Pakdel F. Cyclophosphamide-induced testicular toxicity ameliorate by American ginseng treatment: an experimental study. Int J Reprod Biomed 2018; 16:711-718.
44. Jahan S, Ain QU, Ullah H. Therapeutic effects of quercetin against bisphenol A induced testicular damage in male sprague dawley rats. Syst Biol Reprod Med 2016; 62:114-124.
45. Fadda LM, Attia HA, Al-Rasheed NM, Ali HM, Al-Rasheed NM. Roles of some antioxidants in modulation of cardiac myopathy induced by sodium nitrite via down-regulation of mRNA expression of NF-κB, Bax, and flt-1 and suppressing DNA damage. Saudi Pharm J 2018; 26:217-223.
46. Ebokaiwe AP, Mathur PP, Farombi EO. Quercetin and vitamin E attenuate bonny light crude oil-induced alterations in testicular apoptosis, stress proteins and steroidogenic acute regulatory protein in wistar rats. Drug Chem Toxicol 2016; 39:424-431.
47. Abarikwu SO, Pant AB, Farombi EO. Quercetin decreases steroidogenic enzyme activity, NF-κB expression, and oxidative stress in cultured leydig cells exposed to atrazine. Mol Cell Biochem 2013; 373:19-28.
48. Sanjay S, Girish C, Toi PC, Bobby Z. Quercetin modulates NRF2 and NF-κB/TLR-4 pathways to protect against isoniazid- and rifampicin-induced hepatotoxicity in vivo. Can J Physiol Pharmacol. 2021; 99:952-963.
49. Le X, Luo P, Gu Y, Tao Y, Liu H. Squid ink polysaccharide reduces cyclophosphamide-induced testicular damage via Nrf2/ARE activation pathway in mice. Iran J Basic Med Sci 2015; 18:827-831.
50. Wardyn JD, Ponsford AH, Sanderson CM. Dissecting molecular cross-talk between Nrf2 and NF-κB response pathways. Biochem Soc Trans 2015; 43:621-626.
51. Sivandzade F, Alqahtani F, Sifat A, Cucullo L. The cerebrovascular and neurological impact of chronic smoking on post-traumatic brain injury outcome and recovery: An in vivo study. J Neuroinflammation 2020; 17:133-151.
52. Yardim A, Kandemir FM, Ozdemir S, Kucukler S, Comakli S, Gur C, et al. Quercetin provides protection against the peripheral nerve damage caused by vincristine in rats by suppressing caspase 3, NF-κB, ATF-6 pathways and activating Nrf2, Akt pathways. Neurotoxicology 2020; 81:137-146.
53. Kale J, Osterlund EJ, Andrews DW. BCL-2 family proteins: Changing partners in the dance towards death. Cell Death Differ 2018; 25:65-80.
54. Wang Q, Zhao XF, Ji YL, Wang H, Liu P, Zhang C et al. Mitochondrial signaling pathway is also involved in bisphenol a induced germ cell apoptosis in testes. Toxicol Lett 2010; 199:129-135.
55. Mehrbod P, Ande SR, Alizadeh J, Rahimizadeh S, Shariati A, Malek H et al. The roles of apoptosis, autophagy and unfolded protein response in arbovirus, influenza virus, and HIV infections. Virulence 2019; 10:376-413.
56. Adams JM, Cory S. The Bcl-2 apoptotic switch in cancer development and therapy. Oncogene 2007; 26:1324-1337.
57. Khodabandeh Z, Dolati P, Zamiri MJ, Mehrabani D, Bordbar H, Alaee S, et al. Protective effect of quercetin on testis structure and apoptosis against lead acetate toxicity: an stereological study. Biol Trace Elem Res 2021; 199:3371-3381.
58. Habas K, Brinkworth MH, Anderson D. Diethylstilbestrol induces oxidative DNA damage, resulting in apoptosis of spermatogonial stem cells in vitro. Toxicology 2017; 382:117-121.
59. Wang JY, Nie YX, Dong BZ, Cai ZC, Zeng XK, Du L, et al. Quercetin protects islet β-cells from oxidation-induced apoptosis via Sirt3 in T2DM. Iran J Basic Med Sci 2021; 24:629-635.