Environmental Science and Pollution Research

, Volume 26, Issue 11, pp 11427–11437 | Cite as

Antioxidant and cytoprotective effects of N-acetylcysteine against subchronic oral glyphosate-based herbicide-induced oxidative stress in rats

  • Ruhi TurkmenEmail author
  • Yavuz Osman Birdane
  • Hasan Huseyin Demirel
  • Hidayet Yavuz
  • Mustafa Kabu
  • Sinan Ince
Research Article


It is claimed that oxidative stress has a prominent role in the mechanism of toxic effects formed by glyphosate-based herbicide (GBH) in living systems. A strong thiol compound, N-acetylcysteine (NAC), has antioxidative and cytoprotective properties. The objective in this subchronic toxicity study was to identify the prophylactic effect of NAC over histopathological changes and oxidative stress induced by GBH in blood, renal, liver, cardiac, and brain tissues. A sum of 28 male Wistar rats were divided into four equal groups, each containing 7 rats. During the study, group I (control group) was supplied with normal rodent bait and tap water ad libitum. The applied agents were 160 mg/kg NAC to group II, 375 mg/kg as equivalent to 1/10 of lethal dose 50% (LD50) of GBH to group III, and 160 mg/kg of NAC and 375 mg/kg of GBH together once per day as oral gavage to group IV for 8 weeks. While GBH decreased the levels of GSH in blood, liver, kidney, and brain tissues, it considerably increased malondialdehyde levels. On the contrary, these parameters happened to improve in the group supplied with NAC. Besides, it was seen that NAC was observed to improve the histopathologic changes in rat tissues induced by GBH. It was concluded that NAC protects oxidative stress and tissue damage induced by GBH in blood and tissue and this prophylactic effect could be attributed to its antioxidant and free radical sweeper character.


Glyphosate based-herbicide Glyphosate isopropylamine Subchronic toxicity N-Acetylcysteine Oxidative stress Rat 



A short summary of this study was verbally presented at I. International Congress on Medicinal and Aromatic Plants, 09–12 May 2017, in Konya, Turkey.

Funding information

This study was funded by Afyon Kocatepe University Scientific Research Projects Coordination Department (Project No: 16.VF.11).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest.


  1. Abdel-Daim MM, Dessouki AA, Abdel-Rahman HG, Eltaysh R, Alkahtani S (2019) Hepatorenal protective effects of taurine and N-acetylcysteine against fipronil-induced injuries: the antioxidant status and apoptotic markers expression in rats. Sci Total Environ 650:2063–2073. CrossRefGoogle Scholar
  2. Aebi H (1984) Catalase in vitro. Methods Enzymol 105:121–126. CrossRefGoogle Scholar
  3. Akbel E, Arslan-Acaroz D, Demirel HH, Kucukkurt I, Ince S (2018) The subchronic exposure to malathion, an organophosphate pesticide, causes lipid peroxidation, oxidative stress, and tissue damage in rats: the protective role of resveratrol. Toxicol Res (Camb) 7:503–512. CrossRefGoogle Scholar
  4. Aksoy Y (2002) Antioksidan Mekanizmada Glutatyonun Rolü. T Klin J Med Sci 22:442–448Google Scholar
  5. Alp H, Pinar N, Dokuyucu R, Kaplan I, Sahan M, Senol S, Karakus A, Yaldiz M (2016) Effects of intralipid and caffeic acid phenyl esther (CAPE) against hepatotoxicity and nephrotoxicity caused by glyphosate isopropylamine (GI). Eur J Inflamm 14:3–9. CrossRefGoogle Scholar
  6. Bai SH, Ogbourne SM (2016) Glyphosate: environmental contamination, toxicity and potential risks to human health via food contamination. Environ Sci Pollut Res 23:18988–19001. CrossRefGoogle Scholar
  7. Benedetti AL, Vituri CDL, Trentin AG et al (2004) The effects of sub-chronic exposure of Wistar rats to the herbicide Glyphosate-Biocarb®. Toxicol Lett 153:227–232. CrossRefGoogle Scholar
  8. Beutler E (1963) Improved method for the determination of blood glutathione. J Lab Clin Med 61:882–888Google Scholar
  9. Burella PM, Odetti LM, Simoniello MF, Poletta GL (2018) Oxidative damage and antioxidant defense in Caiman latirostris (broad-snouted caiman) exposed in ovo to pesticide formulations. Ecotoxicol Environ Saf 161:437–443. CrossRefGoogle Scholar
  10. Çaǧlar S, Kolankaya D (2008) The effect of sub-acute and sub-chronic exposure of rats to the glyphosate-based herbicide Roundup. Environ Toxicol Pharmacol 25:57–62. CrossRefGoogle Scholar
  11. Cattaneo R, Clasen B, Loro VL, de Menezes CC, Pretto A, Baldisserotto B, Santi A, de Avila LA (2011) Toxicological responses of Cyprinus carpio exposed to a commercial formulation containing glyphosate. Bull Environ Contam Toxicol 87:597–602. CrossRefGoogle Scholar
  12. Cattani D, de Liz Oliveira Cavalli VL, Heinz Rieg CE, et al (2014) Mechanisms underlying the neurotoxicity induced by glyphosate-based herbicide in immature rat hippocampus: involvement of glutamate excitotoxicity. Toxicology., 320, 34, 45
  13. Çavuşoğlu K, Yapar K, Oruç E, Yalçın E (2011) Protective effect of Ginkgo biloba L. leaf extract against glyphosate toxicity in Swiss albino mice. J Med Food 14:1263–1272. CrossRefGoogle Scholar
  14. Chłopecka M, Mendel M, Dziekan N, Karlik W (2014) Glyphosate affects the spontaneous motoric activity of intestine at very low doses - in vitro study. Pestic Biochem Physiol 113:25–30. CrossRefGoogle Scholar
  15. Dar MA, Sultana M, Mir AH, Bhat MA, Wani TA, Haq Z (2018) Sub-acute oral toxicity of Roundup® and ammonium nitrate with special reference to oxidative stress indices in wistar rats. Indian J Anim Res 52:405–408. Google Scholar
  16. de Souza JS, Kizys MML, da Conceição RR, Glebocki G, Romano RM, Ortiga-Carvalho TM, Giannocco G, da Silva IDCG, Dias da Silva MR, Romano MA, Chiamolera MI (2017) Perinatal exposure to glyphosate-based herbicide alters the thyrotrophic axis and causes thyroid hormone homeostasis imbalance in male rats. Toxicology 377:25–37. CrossRefGoogle Scholar
  17. Dodd S, Dean O, Copolov DL, Malhi GS, Berk M (2008) N -acetylcysteine for antioxidant therapy: pharmacology and clinical utility. Expert Opin Biol Ther 8:1955–1962. CrossRefGoogle Scholar
  18. Drabkin DL, Austin JH (1935) Spectrophotometric studies. J Biol Chem 112:51–65Google Scholar
  19. Draper HH, Hadley M (1990) Malondialdehyde determination as index of lipid peroxidation. Methods Enzymol 186:421–431. CrossRefGoogle Scholar
  20. El-Bini Dhouib I, Annabi A, Jrad A et al (2015) Carbosulfan-induced oxidative damage following subchronic exposure and the protective effects of N-acetylcysteine in rats. Gen Physiol Biophys 34:249–261. CrossRefGoogle Scholar
  21. El-Shenawy NS (2009) Oxidative stress responses of rats exposed to Roundup and its active ingredient glyphosate. Environ Toxicol Pharmacol 28:379–385. CrossRefGoogle Scholar
  22. Eraslan G, Kanbur M, Siliğ Y, Karabacak M, Soyer Sarica Z, Şahin S (2016) The acute and chronic toxic effect of cypermethrin, propetamphos, and their combinations in rats. Environ Toxicol 31:1415–1429. CrossRefGoogle Scholar
  23. Gibson-Corley KN, Olivier AK, Meyerholz DK (2013) Principles for valid histopathologic scoring in research. Vet Pathol 50:1007–1015. CrossRefGoogle Scholar
  24. Gress S, Lemoine S, Séralini GE, Puddu PE (2015) Glyphosate-based herbicides potently affect cardiovascular system in mammals: review of the literature. Cardiovasc Toxicol 15:117–126. CrossRefGoogle Scholar
  25. Gultekin F, Delibas N, Yasar S, Kilinc I (2001) In vivo changes in antioxidant systems and protective role of melatonin and a combination of vitamin C and vitamin E on oxidative damage in erythrocytes induced by chlorpyrifos-ethyl in rats. Arch Toxicol 75:88–96. CrossRefGoogle Scholar
  26. Guyton KZ, Loomis D, Grosse Y, el Ghissassi F, Benbrahim-Tallaa L, Guha N, Scoccianti C, Mattock H, Straif K, International Agency for Research on Cancer Monograph Working Group, IARC, Lyon, France (2015) Carcinogenicity of tetrachlorvinphos, parathion, malathion, diazinon, and glyphosate. Lancet Oncol 16:490–491. CrossRefGoogle Scholar
  27. Hamdaoui L, Naifar M, Mzid M, Ben Salem M, Chtourou A, Makni-Ayadi F, Sahnoun Z, Rebai T (2016) Nephrotoxicity of Kalach 360 SL: biochemical and histopathological findings. Toxicol Mech Methods 26:685–691. CrossRefGoogle Scholar
  28. Hurst GA, Shaw PB, LeMaistre CA (1967) Laboratory and clinical evaluation of the mucolytic properties of acetylcysteine. Am Rev Respir Dis 96:962–970. Google Scholar
  29. Ince S, Kucukkurt I, Demirel HH, Turkmen R, Sever E (2012) Thymoquinone attenuates cypermethrin induced oxidative stress in Swiss albino mice. Pestic Biochem Physiol 104:229–235. CrossRefGoogle Scholar
  30. Ince S, Kucukkurt I, Demirel HH, Turkmen R, Zemheri F, Akbel E (2013) The role of thymoquinone as antioxidant protection on oxidative stress induced by imidacloprid in male and female Swiss albino mice. Toxicol Environ Chem 95:318–329. CrossRefGoogle Scholar
  31. Jasper R, Locatelli GO, Pilati C, Locatelli C (2012) Evaluation of biochemical, hematological and oxidative parameters in mice exposed to the herbicide glyphosate-Roundup®. Interdiscip Toxicol 5:133–140. CrossRefGoogle Scholar
  32. Kamboj A, Kumaresan N, Kiran R, Sandhir R (2006) Neuroprotective potential of N-acetylcysteine in carbofuran neurotoxicity: a biochemical and histopathological study. Toxicol Environ Chem 88:745–753. CrossRefGoogle Scholar
  33. Lajmanovich RC, Attademo AM, Simoniello MF, Poletta GL, Junges CM, Peltzer PM, Grenón P, Cabagna-Zenklusen MC (2015) Harmful effects of the dermal intake of commercial formulations containing chlorpyrifos, 2,4-D, and glyphosate on the common toad Rhinella arenarum (Anura: Bufonidae). Water Air Soil Pollut 226.
  34. Lasram MM, Lamine AJ, Dhouib IB, Bouzid K, Annabi A, Belhadjhmida N, Ahmed MB, el Fazaa S, Abdelmoula J, Gharbi N (2014) Antioxidant and anti-inflammatory effects of N-acetylcysteine against malathion-induced liver damages and immunotoxicity in rats. Life Sci 107:50–58. CrossRefGoogle Scholar
  35. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275Google Scholar
  36. Luck H (1965) Catalase. In: Bergmeyer HU (ed) Methods of enzymatic analysis. Academic Press, New York, pp 885–894.
  37. Mañas F, Peralta L, Raviolo J, Ovando HG, Weyers A, Ugnia L, Cid MG, Larripa I, Gorla N (2009) Genotoxicity of glyphosate assessed by the comet assay and cytogenetic tests. Environ Toxicol Pharmacol 28:37–41. CrossRefGoogle Scholar
  38. Mesnage R, Defarge N, Spiroux de Vendômois J, Séralini GE (2015) Potential toxic effects of glyphosate and its commercial formulations below regulatory limits. Food Chem Toxicol 84:133–153. CrossRefGoogle Scholar
  39. Mohammadi-Sardoo M, Mandegary A, Nabiuni M, Nematollahi-Mahani SN, Amirheidari B (2018) Mancozeb induces testicular dysfunction through oxidative stress and apoptosis: protective role of N-acetylcysteine antioxidant. Toxicol Ind Health 34:798–811. CrossRefGoogle Scholar
  40. Moon JM, Chun BJ, Cho YS, Lee SD, Hong YJ, Shin MH, Jung EJ, Ryu HH (2018) Cardiovascular effects and fatality may differ according to the formulation of glyphosate salt herbicide. Cardiovasc Toxicol 18:99–107. CrossRefGoogle Scholar
  41. Murussi CR, Costa MD, Leitemperger JW, Guerra L, Rodrigues CCR, Menezes CC, Severo ES, Flores-Lopes F, Salbego J, Loro VL (2016) Exposure to different glyphosate formulations on the oxidative and histological status of Rhamdia quelen. Fish Physiol Biochem 42:445–455. CrossRefGoogle Scholar
  42. Ohkawa H, Ohishi N, Yagi K (1979) Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem 95:351–358. CrossRefGoogle Scholar
  43. Pal S, Chatterjee AK (2004) Protective effect of N-acetylcysteine against arsenic-induced depletion in vivo of carbohydrate. Drug Chem Toxicol 27:179–189CrossRefGoogle Scholar
  44. Piola L, Fuchs J, Oneto ML, Basack S, Kesten E, Casabé N (2013) Comparative toxicity of two glyphosate-based formulations to Eisenia andrei under laboratory conditions. Chemosphere. 91:545–551. CrossRefGoogle Scholar
  45. Prescott LF (1983) New approaches in managing drug overdosage and poisoning. Br Med J 287:274–276CrossRefGoogle Scholar
  46. Roy NM, Carneiro B, Ochs J (2016) Glyphosate induces neurotoxicity in zebrafish. Environ Toxicol Pharmacol 42:45–54. CrossRefGoogle Scholar
  47. Samsel A, Seneff S (2013) Glyphosate, pathways to modern diseases II: celiac sprue and gluten intolerance. Interdiscip Toxicol 6:159–184. CrossRefGoogle Scholar
  48. Samsel A, Seneff S (2015) Glyphosate, pathways to modern diseases III: manganese, neurological diseases, and associated pathologies. Surg Neurol Int 6:45. Google Scholar
  49. Sun YI, Oberley L w, Ying L (1988) A simple method for clinical assay of superoxide dismutase. Clin Chem 34:497–500. Google Scholar
  50. Tizhe EV, Ibrahim NDG, Fatihu MY et al (2013) Haematogical changes induced by subchronic glyphosate exposure: ameliorative effect of zinc in Wistar rats. 11:28–35.
  51. Tizhe EV, Ibrahim NDG, Fatihu MY, Onyebuchi II, George BDJ, Ambali SF, Shallangwa JM (2014) Influence of zinc supplementation on histopathological changes in the stomach, liver, kidney, brain, pancreas and spleen during subchronic exposure of Wistar rats to glyphosate. Comp Clin Pathol 23:1535–1543CrossRefGoogle Scholar
  52. USEPA (2016) Report on the 2016 U.S. Environmental Protection Agency (EPA) International Decontamination Research and Development ConferenceGoogle Scholar
  53. Webster TMU, Santos EM (2015) Global transcriptomic profiling demonstrates induction of oxidative stress and of compensatory cellular stress responses in brown trout exposed to glyphosate and Roundup. BMC Genomics 16:32. CrossRefGoogle Scholar
  54. Whitehouse L, Wong L, Paul C (1985) Postabsorption antidotal effects of N-acetylcysteine on acetaminophen-induced hepatotoxicity in the mouse. Can J Physiol Pharmacol 63:431–437CrossRefGoogle Scholar
  55. Winterbourn CC, Hawkins RE, Brian MCR (1975) The estimation of red cell superoxide dismutase activity. J Lab Clin Med 85:337–341Google Scholar
  56. Wunnapuk K, Gobe G, Endre Z, Peake P, Grice JE, Roberts MS, Buckley NA, Liu X (2014) Use of a glyphosate-based herbicide-induced nephrotoxicity model to investigate a panel of kidney injury biomarkers. Toxicol Lett 225:192–200. CrossRefGoogle Scholar
  57. Yarsan E, Tanyuksel M, Celik S, Aydin A (1999) Effects of aldicarb and malathion on lipid peroxidation. Bull Environ Contam Toxicol 63:575–581. CrossRefGoogle Scholar
  58. Youness A, El-Toukhy et al (2016) The protective effect of orange juice on glyphosate toxicity in adult male mice. J Chem Pharm Res 8:13–28Google Scholar
  59. Yurumez Y, Cemek M, Yavuz Y, Birdane YO, Buyukokuroglu ME (2007) Beneficial effect of N-acetylcysteine against organophosphate toxicity in mice. Biol Pharm Bull 30:490–494. CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Pharmacology and Toxicology, Faculty of Veterinary MedicineAfyon Kocatepe UniversityAfyonkarahisarTurkey
  2. 2.Bayat Vocational SchoolAfyon Kocatepe UniversityAfyonkarahisarTurkey
  3. 3.Department of Internal Medicine, Faculty of Veterinary MedicineAfyon Kocatepe UniversityAfyonkarahisarTurkey

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