Environmental Science and Pollution Research

, Volume 24, Issue 1, pp 938–946 | Cite as

Proanthocyanidin protects against acute zearalenone-induced testicular oxidative damage in male mice

  • Miao Long
  • Shuhua Yang
  • Yi Zhang
  • Peng Li
  • Jianxin Han
  • Shuang Dong
  • Xinliang Chen
  • Jianbin HeEmail author
Research Article


Zearalenone (ZEN) exerts a major effect on human and animal health and has led to serious worldwide economic problems. In this study, we investigated whether proanthocyanidin (PC) can prevent ZEN-induced testicular oxidative damage in male mice and explored the underlying mechanism. Kunming mice were injected with ZEN (40 mg kg−1) on the 11th day after intragastric administration of PC (75 or 150 mg/kg) for 10 days; the sperm quality of mice was then analysed statistically. Additionally, testicular morphology parameters related to oxidative damage, apoptosis and the expression of endoplasmic reticulum (ER) stress-related genes (GRP78, CHOP and XBP-1) were all measured. Results showed that ZEN exposure significantly reduced the sperm density, improved the sperm aberration rate, increased the MDA level and reduced SOD and GSH-Px activities. Meanwhile, ZEN was attributed to the downregulation of the expressions of the gene and protein of Bcl-2 and upregulation of the expressions of the gene and protein of Bax and caspase-3. ZEN exposure also upregulated the mRNA expression of GRP78, CHOP and XBP-1; however, PC pre-treatment reduced ZEN-induced oxidative damage and tended to maintain normal testicular morphology. Furthermore, PC pre-treatment substantially downregulated the expressions of the GRP78, CHOP and XBP-1 and upregulated the expression of the Bcl-2 gene. In conclusion, PC, due to its anti-oxidative ability, could ameliorate ZEN-induced testicular reproductive toxicity in male mice by decreasing ER stress and testicular cell apoptosis.


Zearaleone Mice testis Oxidative damage Proanthocyanidin Apoptosis 









Superoxide dismutase


Glutathione peroxidase




Endoplasmic reticulum


Glucose-regulated protein 78


CCAAT/enhancer binding protein homologous protein


X-box binding protein1



This work was financially supported by the National Natural Science Foundation of China (grant no. 31201961; grant no. 31302152). China Postdoctoral Science Foundation funded project grants (2014M551125); The general program of Liaoning Provincial Department of Education Science Research (L2014561).


  1. Abid-Essefi S, Baudrimont I, Hassen W, Ouanes Z, Mobio TA, Anane R (2003) DNA fragmentation, apoptosis and cell cycle arrest induced by zearalenone in cultured DOK, Vero and Caco-2 cells: prevention by vitamin E. Toxicology 192:237–248CrossRefGoogle Scholar
  2. Abid-Essefi S, Zaied C, Bouaziz C, Salem IB, Kaderi R, Bacha H (2012) Protective effect of aqueous extract of Allium sativum against zearalenone toxicity mediated by oxidative stress. Exp Toxicol Pathol 64:689–695CrossRefGoogle Scholar
  3. Aruoma OI, Sun B, Fujii H, Neergheen VS, Bahorun T, Kang KS (2006) Low molecular proanthocyanidin dietary biofactor Oligonol: its modulation of oxidative stress, bioefficacy, neuroprotection, food application and chemoprevention potentials. Biofactors 27:245–265CrossRefGoogle Scholar
  4. Banjerdpongchai R, Kongtawelert P, Khantamat O, Srisomsap C, Chokchaichamnankit D, Subhasitanont P, Svasti J (2010) Mitochondrial and endoplasmic reticulum stress pathways cooperate in zearalenone-induced apoptosis of human leukemic cells. J Hematol Oncol 3:50CrossRefGoogle Scholar
  5. Ben Salah-Abbès J, Abbès S, Ouanes Z, Abdel-Wahhab MA, Bacha H, Oueslati R (2009) Isothiocyanate from the Tunisian radish (Raphanus sativus) prevents genotoxicity of Zearalenone in vivo and in vitro. Mutat Res 677:59–65CrossRefGoogle Scholar
  6. Ben Salem I, Prola A, Boussabbeh M, Guilbert A, Bacha H, Abid-Essefi S (2015a) Crocin and Quercetin protect HCT116 and HEK293 cells from Zearalenone-induced apoptosis by reducing endoplasmic reticulum stress. Cell Stress Chaperones 20:927–938CrossRefGoogle Scholar
  7. Ben Salem I, Boussabbeh M, Helali S, Abid-Essefi S, Bacha H (2015b) Protective effect of Crocin against zearalenone-induced oxidative stress in liver and kidney of Balb/c mice Environ Sci Pollut Res 22:19069–19076Google Scholar
  8. Boeira SP, Filho CB, Del'Fabbro L, Royes LF, Jessé CR, Oliveira MS (2012) Possible role for glutathione-S-transferase in the oligozoospermia elicited by acute zearalenone administration in Swiss albino mice. Toxicon 60:358–366CrossRefGoogle Scholar
  9. Boeira SP, Filho CB, Del'Fabbro L, Roman SS, Royes LF, Fighera MR (2014) Lycopene treatment prevents hematological, reproductive and histopathological damage induced by acute zearalenone administration in male Swiss mice. Exp Toxicol Pathol 66:179–185CrossRefGoogle Scholar
  10. Boeira SP, Funck VR, Borges Filho C, Del'Fabbro L, de Gomes MG, Donato F (2015) Lycopene protects against acute zearalenone-induced oxidative, endocrine, inflammatory and reproductive damages in male mice. Chem Biol Interact 230:50–57CrossRefGoogle Scholar
  11. Caillet S, Salmiéri S, Lacroix M (2006) Evaluation of free radical-scavenging properties of commercial grape phenol extracts by a fast colorimetric method. Food Chem 95:1–8CrossRefGoogle Scholar
  12. Cao AH, Wang J, Gao HQ, Zhang P, Qiu J (2015) Beneficial clinical effects of grape seed proanthocyanidin extract on the progression of carotid atherosclerotic plaques. J Geriatr Cardiol 12:417–423Google Scholar
  13. Chen Q, Zhang R, Li WM, Niu YJ, Guo HC, Liu XH (2013) The protective effect of grape seed procyanidin extract against cadmium-induced renal oxidative damage in mice. Environ Toxicol Pharmacol 36:759–768CrossRefGoogle Scholar
  14. Chen F, Li Q, Zhang Z, Lin P, Lei L, Wang A, Jin Y (2015) Endoplasmic reticulum stress cooperates in zearalenone-induced cell death of RAW 264.7 macrophages. Int J Mol Sci 16:19780–19795CrossRefGoogle Scholar
  15. Chen H, Yang H, Pan L, Wang W, Liu X, Ren X (2016) The molecular mechanisms of XBP-1 gene silencing on IRE1α-TRAF2-ASK1-JNK pathways in oral squamous cell carcinoma under endoplasmic reticulum stress. Biomed Pharmacother 77:108–113CrossRefGoogle Scholar
  16. Dominighini A, Ferrero M, Crosetti D, Ronco MT, Gonzálvez J, Urli L (2015) Effects of proanthocyanidin enriched extract from ligaria cuneifolia on plasma cholesterol and hemorheological parameters in vivo and in vitro studies. Clin Hemorheol Microcirc 60:317–325CrossRefGoogle Scholar
  17. Escrivá L, Font G, Manyes L (2015) In vivo toxicity studies of fusarium mycotoxins in the last decade: a review. Food Chem Toxicol 78:185–206CrossRefGoogle Scholar
  18. Gao Z, Liu G, Hu Z, Li X, Yang X, Jiang B, Li X (2014) Grape seed proanthocyanidin extract protects from cisplatin-induced nephrotoxicity by inhibiting endoplasmic reticulum stress-induced apoptosis. Mol Med Rep 9:801–807Google Scholar
  19. Hala AHK, Inas ZAA, Gehan MK (2010) Grape seed extract alleviate reproductive toxicity caused by aluminium chloride in male rats. J Am Sci 6:1200–1209Google Scholar
  20. Hu J, Xu M, Dai Y, Ding X, Xiao C, Ji H, Xu Y (2016) Exploration of Bcl-2 family and caspases-dependent apoptotic signaling pathway in Zearalenone-treated mouse endometrial stromal cells. Biochem Biophys Res Commun 476:553–559CrossRefGoogle Scholar
  21. JECFA (2000) Zearalenone, in: Joint FAO/WHO Expert Committee on Food Additives (Ed), Safety Evaluation of Certain Food Additives and Contaminants WHO/FAO Food Additives Series 44 IPCS -International Programme on Chemical Safety. WHO, GenevaGoogle Scholar
  22. Kim IH, Son HY, Cho SW, Ha CS, Kang BH (2003) Zearalenone induces male germ cell apoptosis in rats. Toxicol Lett 138:185–192CrossRefGoogle Scholar
  23. Li J, Wang JJ, Yu Q, Wang M, Zhang SX (2009) Endoplasmic reticulum stress is implicated in retinal inflammation and diabetic retinopathy. FEBS Lett 583:1521–1527CrossRefGoogle Scholar
  24. Li Y, He X, Yang X, Huang K, Luo Y, Zhu L (2015a) Zinc inhibits the reproductive toxicity of Zearalenone in immortalized murine ovarian granular KK-1 cells. Sci Rep 5:14277CrossRefGoogle Scholar
  25. Li SG, Ding YS, Niu Q, Xu SZ, Pang LJ, Ma RL (2015b) Grape seed proanthocyanidin extract alleviates arsenic-induced oxidative reproductive toxicity in male mice. Biomed Environ Sci 28:272–280CrossRefGoogle Scholar
  26. Liang Z, Ren Z, Gao S, Chen Y, Yang Y, Yang D (2015) Individual and combined effects of deoxynivalenol and zearalenone on mouse kidney. Environ Toxicol Pharmacol 40:686–691CrossRefGoogle Scholar
  27. Lin P, Chen F, Sun J, Zhou J, Wang X, Wang N, Li X, Zhang Z, Wang A, Jin Y (2015) Mycotoxin zearalenone induces apoptosis in mouse Leydig cells via an endoplasmic reticulum stress-dependent signalling pathway. Reprod Toxicol 52:71–77CrossRefGoogle Scholar
  28. Mouradov A, Spangenberg G (2014) Flavonoids: a metabolic network mediating plants adaptation to their real estate. Front Plant Sci 5:620. doi: 10.3389/fpls.2014.00620 CrossRefGoogle Scholar
  29. Nie C, Zhou J, Qin X, Shi X, Zeng Q, Liu J (2016) Reduction of apoptosis by proanthocyanidin-induced autophagy in the human gastric cancer cell line MGC-803. Oncol Rep 35:649–658Google Scholar
  30. Pintha K, Yodkeeree S, Limtrakul P (2015) Proanthocyanidin in red rice inhibits MDA-MB-231 breast cancer cell invasion via the expression control of invasive proteins. Biol Pharm Bull 38:571–581CrossRefGoogle Scholar
  31. Prasad R, Vaid M, Katiyar SK (2012) Grape proanthocyanidin inhibit pancreatic cancer cell growth in vitro and in vivo through induction of apoptosis and by targeting the PI3K/Akt pathway. PLoS One 7(8):e43064. doi: 10.1371/journal.pone.0043064 CrossRefGoogle Scholar
  32. Ren ZH, Deng HD, Wang YC, Deng JL, Zuo ZC, Wang Y (2016) The Fusarium toxin zearalenone and deoxynivalenol affect murine splenic antioxidant functions, interferon levels, and T-cell subsets. Environ Toxicol Pharmacol 41:195–200CrossRefGoogle Scholar
  33. Ron D, Habener JF (1992) CHOP, a novel developmentally regulated nuclear protein that dimerizes with transcription factors C/EBP and LAP and functions as a dominant-negative inhibitor of gene transcription. Gene Dev 6:439–453CrossRefGoogle Scholar
  34. Salem IB, Boussabbeh M, Neffati F, Najjar MF, Abid-Essefi S, Bacha H (2016) Zearalenone-induced changes in biochemical parameters, oxidative stress and apoptosis in cardiac tissue: protective role of crocin. Hum Exp Toxicol 35:623–634CrossRefGoogle Scholar
  35. Su L, Deng Y, Zhang Y, Li C, Zhang R, Sun Y (2011) Protective effects of grape seed procyanidin extract against nickel sulfate-induced apoptosis and oxidative stress in rat testes. Toxicol Mech Methods 21:487–494CrossRefGoogle Scholar
  36. Venkataramana M, Chandra Nayaka S, Anand T, Rajesh R, Aiyaz M, Divakara ST (2014) Zearalenone induced toxicity in SHSY-5Y cells: the role of oxidative stress evidenced by N-acetyl cysteine. Food and chemical toxicology: an international journal published for the British Industrial Biological Research Association 65:335–342CrossRefGoogle Scholar
  37. Wang Y, Zheng W, Bian X, Yuan Y, Gu J, Liu X (2014) Zearalenone induces apoptosis and cytoprotective autophagy in primary Leydig cells. Toxicol Lett 226:182–191CrossRefGoogle Scholar
  38. Yang JY, Wang GX, Liu JL, Fan JJ, Cui S (2007) Toxic effects of zearalenone and its derivatives alpha-zearalenol on male reproductive system in mice. Reprod Toxicol 4:381–387CrossRefGoogle Scholar
  39. Zhang Z, Zheng L, Zhao Z, Shi J, Wang X, Huang J (2014) Grape seed proanthocyanidins inhibit H2O2-induced osteoblastic MC3T3-E1 cell apoptosis via ameliorating H2O2-induced mitochondrial dysfunction. J Toxicol Sci 39:803–813CrossRefGoogle Scholar
  40. Zhang JQ, Xing BS, Zhu CC, Shen M, Yu FX, Liu HL (2015) Protective effect of proanthocyanidin against oxidative ovarian damage induced by 3-nitropropionic acid in mice. Genet Mol Res 14:2484–2494CrossRefGoogle Scholar
  41. Zhao YM, Gao LP, Zhang HL, Guo JX, Guo PP (2014) Grape seed proanthocyanidin extract prevents DDP-induced testicular toxicity in rats. Food Funct 5:605–611CrossRefGoogle Scholar
  42. Zhou C, Zhang Y, Yin S, Jia Z, Shan A (2015) Biochemical changes and oxidative stress induced by zearalenone in the liver of pregnant rats. Hum Exp Toxicol 34:65–73CrossRefGoogle Scholar
  43. Zourgui L, Golli EE, Bouaziz C, Bacha H, Hassen W (2008) Cactus (Opuntia ficus-indica) cladodes prevent oxidative damage induced by the mycotoxin zearalenone in Balb/C mice. Food Chem Toxico l46:1817–1824CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  • Miao Long
    • 1
  • Shuhua Yang
    • 1
  • Yi Zhang
    • 1
  • Peng Li
    • 1
  • Jianxin Han
    • 1
  • Shuang Dong
    • 1
  • Xinliang Chen
    • 1
  • Jianbin He
    • 1
    Email author
  1. 1.Key Laboratory of Zoonosis of Liaoning Province, College of Animal Science and Veterinary MedicineShenyang Agricultural UniversityShenyangChina

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