Protective effects of naringin against oxaliplatin-induced testicular damage in rats: Involvement of oxidative stress, inflammation, endoplasmic reticulum stress, apoptosis, and histopathology

Document Type : Original Article


1 Department of Histology and Embryology, Faculty of Medicine, Aksaray University, Aksaray, Turkey

2 Department of Biochemistry, Faculty of Veterinary Medicine, Atatürk University, Erzurum, Turkey

3 Department of Medical Biochemistry, Faculty of Medicine, Bilecik Seyh Edebali University, Bilecik, Turkey

4 Department of Medical Biochemistry, Faculty of Medicine, Aksaray University, Aksaray, Turkey


Objective(s): Oxaliplatin (OXL) is a platinum-based chemotherapeutic agent widely used in the treatment of colorectal cancer. Unfortunately, this important drug also causes unwanted side effects such as neuropathy, ototoxicity, and testicular toxicity. This study aimed to investigate the possible protective effects of naringin (NRG) against OXL-induced testicular toxicity in rats. 
Materials and Methods: In the present study, rats were injected with OXL (4 mg/kg, b.w./day, IP) in 5% dextrose solution 30 min after oral administration of NRG (50 and 100 mg/kg, b.w./day) on the 1st, 2nd, 5th, and 6th days. Then, the rats were sacrificed on the 7th day and the testicular tissues were removed. 
Results: The results showed that NRG decreased (P<0.001) lipid peroxidation, increased (P<0.001) the activities of superoxide dismutase (SOD), glutathione peroxidase (GPx), catalase (CAT), and the levels of glutathione (GSH), and also maintained the testis histological architecture and integrity. NRG decreased the levels of apoptosis-related markers such as caspase-3, Bax, and Apaf-1 and increased Bcl2 in the OXL-induced testicular toxicity (P<0.001). In addition, NRG reversed the changes in mRNA transcript levels of oxidative stress, inflammation, and endoplasmic reticulum stress parameters such as Nrf2, HO-1, NQO1, RAGE, NLRP3, MAPK-14, STAT3, NF-κB, IL-1β, TNF-α, PERK, IRE1, ATF6, and GRP78 in OXL-induced testicular toxicity (P<0.001). 
Conclusion: Our results demonstrated that NRG can protect against OXL-induced testicular toxicity by enhancing the anti-oxidant defense system and suppressing apoptosis, inflammation, and endoplasmic reticulum stress.


Main Subjects

1. Famurewa AC, Mukherjee AG, Wanjari UR,  Sukumar A, Murali R, Renu K, et al. Repurposing FDA-approved drugs against the toxicity of platinum-based anticancer drugs. Life Sci 2022;305:120789.
2. Osman S, Raza A, Al-Zaidan L, Inchakalody VP, Merhi M, Prabhu KS, et al. Dermime, anti-cancer effects of tranilast: An update, biomed. Pharmacother 2021;141:111844.
3. Thurston DE, Pysz I. Chemistry and Pharmacology of Anticancer Drugs. CRC press 2nd ed 2021.
4. Mansoori B, Mohammadi A, Davudian S, Shirjang S, Baradaran B. The different mechanisms of cancer drug resistance: A brief review. Adv Pharm Bull 2017;7: 339-348.
5. Rottenberg S, Disler C, Perego P. The rediscovery of platinum-based cancer therapy. Nat Rev Cancer 2021;21:37-50.
6. Velasco R, Alemany M, Villagrán M, Argyriou AA. Predictive biomarkers of oxaliplatin-induced peripheral neurotoxicity. J Pers Med 2021;11:669-689.
7. Erdem G, Dogan M, Demirci N, Zengin N. Oxaliplatin-induced acute thrombocytopenia. J Cancer Res Ther 2016; 12:509-514.
8. Varışlı B, Caglayan C, Kandemir FM, Gür C, Ayna A, Genç A, et al. Chrysin mitigates diclofenac-induced hepatotoxicity by modulating oxidative stress, apoptosis, autophagy and endoplasmic reticulum stress in rats. Mol Biol Rep 2023: 50: 433-442.
9. Kucukler S, Darendelioğlu E, Caglayan C, Ayna A, Yıldırım S, Kandemir FM. Zingerone attenuates vancomycin-induced hepatotoxicity in rats through regulation of oxidative stress, inflammation and apoptosis. Life Sci 2020; 259:118382.
10. Taysi S, Algburi FS, Taysi ME, Caglayan C. Caffeic acid phenethyl ester: A review on its pharmacological importance, and its association with free radicals, COVID-19, and radiotherapy. Phytother Res 2023; 37:1115-1135.
11. Rauf A, Shariati MA, Imran M, Bashir K, Khan SA, Mitra S, et al. Comprehensive review on naringenin and naringin polyphenols as a potent anticancer agent. Environ Sci Pollut Res 2022; 29:31025-31041.
12. Zeng W, Jin L, Zhang F, Zhang C, Liang W. Naringenin as a potential immunomodulator in therapeutics. Pharmacol Res 2018;135:122-126.
13. Jin L, Zeng W, Zhang F, Zhang C, Liang W. Naringenin ameliorates acute inflammation by regulating intracellular cytokine degradation. J Immun 2017;199:3466-3477.
14. Kandemir FM, Kucukler S, Caglayan C, Gur C, Batil AA, Gülçin İ. Therapeutic effects of silymarin and naringin on methotrexate-induced nephrotoxicity in rats: Biochemical evaluation of anti-inflammatory, antiapoptotic, and anti-autophagic properties. J Food Biochem 2017; 41:e12398.
15. Memariani Z, Abbas SQ, ul Hassan SS, Ahmadi A, Chabra A. Naringin and naringenin as anticancer agents and adjuvants in cancer combination therapy: Efficacy and molecular mechanisms of action, a comprehensive narrative review. Pharmacol Res 2021; 171: 105264.
16. Thangavel P, Vaiyapuri M. Antiproliferative and apoptotic effects of naringin on diethylnitrosamine induced hepatocellular carcinoma in rats. Biomed Aging Pathol 2013; 3:59-64.
17. Caglayan C, Temel Y, Kandemir FM, Yildirim S, Kucukler S. Naringin protects against cyclophosphamide-induced hepatotoxicity and nephrotoxicity through modulation of oxidative stress, inflammation, apoptosis, autophagy, and DNA damage. Environ Sci Pollut Res 2018; 25:20968-20984.
18. Celik H, Kucukler S, Ozdemir S, Comakli S, Gur C, Kandemir FM, et al. Lycopene protects against central and peripheral neuropathy by inhibiting oxaliplatin-induced ATF-6 pathway, apoptosis, inflammation and oxidative stress in brains and sciatic tissues of rats. NeuroToxicology 2020; 80:29-40.
19. Caglayan C, Kandemir FM, Darendelioğlu E, Yıldırım S, Kucukler S, Dortbudak MB. Rutin ameliorates mercuric chloride-induced hepatotoxicity in rats via interfering with oxidative stress, inflammation and apoptosis. J Trace Elem Med Biol 2019; 56:60-68.
20. Sun Y, Oberley LW, Li Y. A simple method for clinical assay of superoxide dismutase, Clin Chem1988; 34:497-500.
21. Aebi H. Catalase in vitro. Methods Enzymol 1984; 105:121-126.
22. Lawrence RA, Burk RF. Glutathione peroxidase activity in selenium-deficient rat liver. Biochem Biophys Res Commun 1976; 71:952-958.
23. Sedlak J, Lindsay RH. Estimation of total, protein-bound, and nonprotein sulfhydryl groups in tissue with Ellman’s reagent. Anal Biochem 1968;25:192-205.
24. Placer ZA, Cushman LL, Johnson BC. Estimation of product of lipid peroxidation (malonyl dialdehyde) in biochemical systems. Anal Biochem 1966; 16;359-364.
25. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ, Protein measurement with the Folin phenol reagent. J Biol Chem 1951; 193:265-275.
26. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 2001;25:402-408.
27. 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.
28. Esteban-Fernández D, Verdaguer JM, Ramírez-Camacho R, Palacios MA, Gómez-Gómez MM. Accumulation, fractionation, and analysis of platinum in toxicologically affected tissues after cisplatin, oxaliplatin, and carboplatin administration. J Anal Toxicol 2008; 32:140-146.
29. Starobova H, Vetter I. Pathophysiology of chemotherapy-induced peripheral neuropathy. Front Mol Neurosci 2017; 10:174-194.
30. Fong CW. Platinum anti-cancer drugs: Free radical mechanism of Pt-DNA adduct formation and anti-neoplastic effect. Free Radic Biol Med 2016;95:216-229.
31. Akaras N, Ileriturk M, Gur C, Kucukler S, Oz M, Kandemir FM. The protective effects of chrysin on cadmium-induced pulmonary toxicity; a multi-biomarker approach. Environ Sci Pollut Res Int 2023;30:89479-89494. 
32. Şimşek H, Akaras N, Gür C, Küçükler S, Kandemir FM. Beneficial effects of chrysin on cadmium-induced nephrotoxicity in rats: Modulating the levels of Nrf2/HO-1, RAGE/NLRP3, and caspase-3/Bax/Bcl-2 signaling pathways. Gene 2023;875:147502. 
33. Gur C, Kandemir FM, Caglayan C, Satıcı E. Chemopreventive effects of hesperidin against paclitaxel-induced hepatotoxicity and nephrotoxicity via amendment of Nrf2/HO-1 and caspase-3/Bax/Bcl-2 signaling pathways. Chem Biol Interact 2022;365:110073. 
34. Cecerska-Heryć E, Surowska O, Heryć R, Serwin N, Napiontek-Balińska S, Dołęgowska B. Are anti-oxidant enzymes essential markers in the diagnosis and monitoring of cancer patients. Clin Biochem 2021; 93:1-8.
35. Guillaumot MA, Cerles O, Bertrand HC, Benoit E, Nicco C, Chouzenoux S, et al. Oxaliplatin-induced neuropathy: The preventive effect of a new super-oxide dismutase modulator. Oncotarget 2019; 10:6418-6431.
36. Cersosimo RJ. Oxaliplatin-associated neuropathy: A review. Ann Pharmacother 2005; 39:128-135.
37. Elsawy H, Alzahrani AM, Alfwuaires M, Abdel-Moneim AM, Khalil M. Beneficial role of naringin against methotrexate-induced injury to rat testes: Biochemical and ultrastructural analyses. Redox Report 2022;27:158-166.
38. Saha S, Buttari B, Panieri E, Profumo E, Saso L. An overview of Nrf2 signaling pathway and its role in inflammation. Molecules 2020; 25:5474-5504.
39. Baird L, Yamamoto M. The molecular mechanisms regulating the KEAP1-NRF2 pathway. Mol Cell Biol 2020; 40:e00099-20.
40. Zangui M, Atkin SL, Majeed M, Sahebkar A. Current evidence and future perspectives for curcumin and its analogues as promising adjuncts to oxaliplatin: state-of-the-art. Pharmacol Res 2019; 141:343-356.
41. Somade OT, Ajayi BO, Adeyi OE, Aina BO, David BO. Sodiya ID. Activation of NF-kB mediates up-regulation of cerebellar and hypothalamic pro-inflammatory chemokines (RANTES and MCP-1) and cytokines (TNF-α, IL-1β, IL-6) in acute edible camphor administration. Scientific African 2029; 5:e00114.
42. Hernández-Aquino E, Muriel P. Beneficial effects of naringenin in liver diseases: Molecular mechanisms. World J Gastroenterol 2018; 24:1679-1707.
43. Tutunchi H, Naeini F, Ostadrahimi A, Hosseinzadeh-Attar MJ. Naringenin, a flavanone with antiviral and anti-inflammatory effects: A promising treatment strategy against COVID-19. Phytother Res 2020; 34:3137-3147.
44. Kelley N, Jeltema D, Duan Y, He Y. The NLRP3 inflammasome: An overview of mechanisms of activation and regulation. Int J Mol Sci 2019; 20: 3328-3351.
45. Jiang X, Wang X. Cytochrome C-mediated apoptosis. Annu Rev Biochem 2004; 73: 87-106.
46. Shakeri R, Kheirollahi A, Davoodi J. Contribution of Apaf-1 to the pathogenesis of cancer and neurodegenerative diseases. Biochimie 2021;190:91-110.
47. Şimşek H, Küçükler S, Gür C, Akaras N, Kandemir FM. Protective effects of sinapic acid against lead acetate-induced nephrotoxicity: a multi-biomarker approach. Environ Sci Pollut Res Int 2023;30:101208-101222.
48. Hu YS, Han X, Liu XH. STAT3: A potential drug target for tumor and inflammation. Curr Top Med Chem 2019; 19:1305-1317.
49. Sparvero LJ, Asafu-Adjei D, Kang R, Tang D, Amin N, Im J, R, et al. RAGE (receptor for advanced glycation endproducts), RAGE ligands, and their role in cancer and inflammation. J Transl Med 2009; 7:17-37.
50.Chen MC, Lin JA, Lin HT, Chen SY, Yen GC. Potential effect of advanced glycation end products (AGEs) on spermatogenesis and sperm quality in rodents. Food  Funct 2019;10:3324-3333.
51. Omolaoye TS, du Plessis SS. Male infertility: A proximate look at the advanced glycation end products. Reprod Toxicol 2020; 93:169-177.
52. Madkour MM, Anbar HS, El-Gamal MI. Current status and future prospects of p38α/MAPK14 kinase and its inhibitors. Eur J Med 2021; 213:113216.
53. Gur C, Kandemir O, Kandemir FM. Investigation of the effects of hesperidin administration on abamectin-induced testicular toxicity in rats through oxidative stress, endoplasmic reticulum stress, inflammation, apoptosis, autophagy, and JAK2/STAT3 pathways. Environ Toxicol 2022; 37:401-412.
54. Akaras N, Gur C, Kucukler S, Kandemir FM. Zingerone reduces sodium arsenite-induced nephrotoxicity by regulating oxidative stress, inflammation, apoptosis and histopathological changes. Chem Biol Interact 2023;374:110410. 
55. Gur C, Akarsu SA, Akaras N, Tuncer SC, Kandemir FM. Carvacrol reduces abnormal and dead sperm counts by attenuating sodium arsenite-induced oxidative stress, inflammation, apoptosis, and autophagy in the testicular tissues of rats. Environ Toxicol. 2023;38:1265-1276. 
56. Dandekar A, Mendez R, Zhang K. Cross talk between ER stress, oxidative stress, and inflammation in health and disease. Methods Mol Biol 2015; 205-214.
57. Hu H, Tian M, Ding C, Yu S. The C/EBP homologous protein (CHOP) transcription factor functions in endoplasmic reticulum stress-induced apoptosis and microbial infection. Front Immunol 2019; 9:3083-3095.
58. Ileriturk M, Kandemir O, Akaras N, Simsek H, Genc A, Kandemir FM. Hesperidin has a protective effect on paclitaxel-induced testicular toxicity through regulating oxidative stress, apoptosis, inflammation and endoplasmic reticulum stress. Reprod Toxicol 2023;118:108369.