Toxicological Research

Korean Society of Toxicology & Korea Environmental Mutagen Society (한국독성학회)
권호 표지
DOI QR코드

DOI QR Code

α-lipoic acid protects testis and epididymis against linuron-induced oxidative toxicity in adult rats

  • Prathima, P. (Department of Biotechnology, Vikrama Simhapuri University) ;
  • Venkaiah, K. (Department of Biotechnology, Vikrama Simhapuri University) ;
  • Daveedu, T. (Department of Biotechnology, Vikrama Simhapuri University) ;
  • Pavani, R. (Department of Biotechnology, Vikrama Simhapuri University) ;
  • Sukeerthi, S. (Department of Biotechnology, Vikrama Simhapuri University) ;
  • Gopinath, M. (Department of Pharmacy, Ratnam Pharmacy College) ;
  • Sainath, Sri Bhashaym (Department of Biotechnology, Vikrama Simhapuri University)
  • Received : 2019.09.29
  • Accepted : 2019.12.10
  • Published : 2020.10.15
⟨ Previous Next ⟩

Abstract

Linuron is well known for its antiandrogenic property. However, the effects of linuron on testicular and epididymal pro- and antioxidant status are not well defined. On the other hand, α-lipoic acid is well known as universal antioxidant. Therefore, the purpose of this study was twofold: firstly to investigate whether linuron exposure alters antioxidant status in the testis and epididymis of rats and if so, whether the supplementation of α-lipoic acid mitigates linuron-induced oxidative toxicity in rats. To address this question, α-lipoic acid at a dose of 70 mg/Kg body weight (three times a week) was administered to linuron exposed rats (10 or 50 mg/Kg body weight, every alternate day over a period of 60 days), and the selected reproductive endpoints were analyzed after 60 days. Respective controls were maintained in parallel. Linuron at selected doses reduced testicular daily sperm count, and epididymal sperm count, sperm motility, sperm viability, and number of tail coiled sperm, reduced activity levels of 3β- and 17β-hydroxysteroid dehydrogenases, decreased expression levels of StAR mRNA, inhibition of testosterone levels, and elevated levels of testicular cholesterol in rats over controls. Linuron intoxication deteriorated the structural integrity of testis and epididymis associated with reduced the reproductive performance over controls. Conversely, α-lipoic acid supplementation enhanced sperm quality and improved the testosterone synthesis pathway in linuron exposed rats over its respective control. Administration of α-lipoic acid restored inhibition of testicular and epididymal enzymatic (superoxide dismutase, catalase, glutathione reductase, glutathione peroxidise) and non-enzymatic (glutathione content), increased lipid peroxidation and protein carbonyl content produced by linuron in rats. α-lipoic acid supplementation inhibited the expression levels of testicular caspase-3 mRNA levels and also its activity in linuron treated rats. To summate, α-lipoic acid-induced protection of reproductive health in linuron treated rats could be attributed to its antioxidant, and steroidogenic properties.

Keywords

Acknowledgement

We thank the Head, Dept. of Biotechnology, VSU, Nellore, AP, India for providing laboratory space and allowed us to utilize the equipments purchased under DST-FIST programme, New Delhi. We thank the Head, Department of Genetics, Narayana Medical College, Nellore for providing animals and the Dr. M. Gobinath, Ratnam Pharmacy College, Muthurkur for providing the animal house facilities. Our special thanks to the Head, Department of Marine Biology for allowing us to utilize ELISA Microplate Reader.

References

  1. EFSA (European Food Safety Authority) (2016) Conclusion on the peer review of the pesticide risk assessment of the active substance linuron. EFSA J 14:4518
  2. Gupta PK (2018) Herbicides and fungicides. In: Veterinary toxicology, basic and clinical principle, 3rd edn. pp 553-567
  3. Gatidou G, Thomaidis NS, Stasinakis AS, Lekkas TD (2007) Simultaneous determination of the endocrine disrupting compounds nonylphenol, nonylphenol ethoxylates, triclosan and bisphenol A in wastewater and sewage sludge by gas chromatography-mass spectrometry. J Chromatogr A 1138:32-41 https://doi.org/10.1016/j.chroma.2006.10.037
  4. Pest management regulatory agency (2012) Proposed re-evaluation decision PRVD2012-02 linuron. Tecnical report prepared for pest management regulatory agency. Health Canada: Ottawa Ontario Canada
  5. Kojima H, Katsura E, Takeuchi S, Niiyama K, Kobayashi K (2004) Screening for estrogen and androgen receptor activities in 200 pesticides by in vitro reporter gene assays using Chinese hamster ovary cells. Environ Health Perspect 112:524-531 https://doi.org/10.1289/ehp.6649
  6. Orton F, Lutz I, Kloas W, Routledge EJ (2009) Endocrine disrupting effects of herbicides and pentachlorophenol: in vitro and in vivo evidence. Environ Sci Technol 43:2144-2150 https://doi.org/10.1021/es8028928
  7. Freyberger A, Witters H, Weimer M, Ahr HJ (2010) Screening for (anti) androgenic properties using a standard operation protocol based on the human stably transfected androgen sensitive PALM cell line first steps towards validation. Reprod Toxicol 30:9-17 https://doi.org/10.1016/j.reprotox.2009.10.002
  8. Wilson VS, Lambright CR, Furr JR, Howdeshell KL, Earl Gray L (2009) The herbicide linuron reduces testosterone production from the fetal rat testis during both in utero and in vitro exposures. Toxicol Lett 186:73-77 https://doi.org/10.1016/j.toxlet.2008.12.017
  9. Uren TM, Perry MH, Santos EM (2015) The herbicide linuron inhibits cholesterol biosynthesis and induces cellular stress responses in brown trout. Environ Sci Technol 49:3110-3118 https://doi.org/10.1021/es505498u
  10. Lambright C, Ostby J, Bobseine K, Wilson V, Hotchkiss AK, Mann PC, Gray LE (2000) Cellular and molecular mechanisms of action of linuron: an antiandrogenic herbicide that produces reproductive malformations in male rats. Toxicol Sci 56:389-399 https://doi.org/10.1093/toxsci/56.2.389
  11. McIntyre BS, Barlow NJ, Wallace DG, Maness SC, Gaido KW, Foster PM (2000) Effects of in utero exposure to linuron on androgen-dependent reproductive development in the male Crl:CD(SD)BR rat. Toxicol Appl Pharmacol 167:87-99 https://doi.org/10.1006/taap.2000.8998
  12. Waller CL, Uma BW, Gray LE Jr, Kelce WR (1996) Three dimensional quantitative structure-activity relationships for androgen receptor ligands. Toxicol Appl Pharmacol 137:219-227 https://doi.org/10.1006/taap.1996.0075
  13. Bauer ER, Meyer HH, Stahischmidt-Allner P, Sauerwein H (1998) Application of an androgen receptor assay for the characterisation of the androgenic or antiandrogenic activity of various phenylurea herbicides and their derivatives. Analyst 123:2485-2487 https://doi.org/10.1039/a804606i
  14. Bai J, Han H, Wang F, Ding H, Hu X, Hu B, Li H, Zheng W, Li Y (2017) Maternal linuron exposure alters testicular development in male offspring rats at the whole genome level. Toxicology 389:13-20 https://doi.org/10.1016/j.tox.2017.07.005
  15. Ding HW, Zheng W, Han H, Hu X, Hu B, Wang F, Su L, Li H, Li Y (2017) Reproductive toxicity of linuron following gestational exposure in rats and underlying mechanisms. Toxicol Lett 266:49-55 https://doi.org/10.1016/j.toxlet.2016.12.013
  16. Hotchkiss AK, Parks-Saldutti LG, Ostby JS, Lambright C, Furr J, Vandenbergh JG, Gray LE (2004) A mixture of the antiandrogens linuron and butyl benzyl phthalate alters sexual differentiation of the male rat in a cumulative fashion. Biol Reprod 71:1852-1861 https://doi.org/10.1095/biolreprod.104.031674
  17. Santos SM, Videira RA, Fernandes MA, Santos MS, Moreno AJ, Vicente JA, Jurado AS (2014) Toxicity of the herbicide linuron as assessed by bacterial and mitochondrial model systems. Toxicol In Vitro 28:932-939 https://doi.org/10.1016/j.tiv.2014.04.004
  18. Nam RK, Hang W, Siminovitch K, Shlien A, Kattan MW, Klotz LH, Trachtenberg J, Lu Y, Zhang J, Yu C, Toi A, Loblaw DA, Venkateswaran V, Stanimirovic A, Sugar L, Malkin D, Seth A, Narod SA (2011) New variants at 10q26 and 15q21 are associated with aggressive prostate cancer in a genome-wide association study from a prostate biopsy screening cohort. Cancer Biol Ther 12:997-1004 https://doi.org/10.4161/cbt.12.11.18366
  19. Bilska A, Wodek L (2005) Lipoic acid - the drug of the future? Pharmacol Rep 57:570-577
  20. El-Bishbishy HA, Aly HA, El-Shafey M (2014) Lipoic acid mitigates bisphenol A induced testicular mitochondrial toxicity in rats. Environ Occup Health 29:875-887
  21. Gules O, Eren U (2016) Protective role of alpha lipoic acid against polychlorobiphenyl (Aroclor 1254)-induced testicular toxicity in rats. Rom J Morph Embryol 57:451-459
  22. Prathima P, Venkaiah K, Pavani R, Daveedu T, Munikumar M, Gobinath M, Valli M, Sainath SB (2017) α-lipoic acid inhibits oxidative stress in testis and attenuates testicular toxicity in rats exposed to carbimazole during embryonic period. Toxicol Rep 4:373-381 https://doi.org/10.1016/j.toxrep.2017.06.009
  23. Prathima P, Pavani R, Sukeerthi S, Sainath SB (2018) α-Lipoic acid inhibits testicular and epididymal oxidative damage and improves fertility efficacy in arsenic-intoxicated rats. J Biochem Mol Toxicol. https://doi.org/10.1002/jbt.22016
  24. Koga T, Ishida T, Takeda T, Ishii Y, Uchi H, Tsukimori K, Yamamot M, Himeno M, Furue M, Yamada H (2012) Restoration of dioxin-induced damage to fetal steroidogenesis and gonadotropin formation by maternal co-treatment with α-lipoic acid. PLoS ONE 7:e40322 https://doi.org/10.1371/journal.pone.0040322
  25. CPCSEA (2003) Committee for the Purpose of control and supervision on experiments on animals PCSEA guidelines for laboratory animal facility. Indian J Pharmacol 35:257-274
  26. Dixit S, Dhar P, Mehra RD (2015) Alpha lipoic acid (ALA) modulates expression of apoptosis associated proteins in hippocampus of rats exposed during postnatal period to sodium arsenite (NaAsO2). Toxicol Rep 2:78-87 https://doi.org/10.1016/j.toxrep.2015.01.011
  27. O'Connor TG, Heron J, Golding J, Beveridge M, Glover V (2002) Maternal antenatal anxiety and children's behavioural emotional problems at 4 years. Br J Psychiatry 180:502-508 https://doi.org/10.1192/bjp.180.6.502
  28. Hayes W, Kruger CL (2014) Hayes' principles and methods of toxicology, 6th edn. CRC Press, Taylor and Francis Group, Boca Raton, Florida
  29. Belsey MA, Moghissi KS, Eliasson R, Paulsen CA, Gallegos AJ, Prasad MR (1980) Laboratory manual for the examination of human semen and semen cervical mucus interaction WHO 1211 Geneva-27 Switzerland
  30. Talbot P, Chacon RS (1981) A triple-stain technique for evaluating normal acrosome reactions of human sperm. J Exp Zool 215:201-208 https://doi.org/10.1002/jez.1402150210
  31. Jeyendran RS, Van der Ven HH, Zaneveld LJD (1992) The hypoosmotic swelling test: an update. Arch Androl 29:105-116 https://doi.org/10.3109/01485019208987714
  32. Ramu S, Jeyendran RS (2013) The hypo-osmotic swelling test for evaluation of sperm membrane integrity. Methods Mol Biol 927:21-25 https://doi.org/10.1007/978-1-62703-038-0_3
  33. Robb GW, Amann RP, Killian GJ (1978) Daily sperm production and epididymal sperm reserves of pubertal and adult rats. J Reprod Fertil 54:103-107 https://doi.org/10.1530/jrf.0.0540103
  34. Linder RE, Strader LF, Slott VL, Suarez JD (1992) Endpoints of spermatotoxicity in the rat after short duration exposures to fourteen reproductive toxicants. Reprod Toxicol 6:491-505 https://doi.org/10.1016/0890-6238(92)90034-Q
  35. Miranda-Spooner M, Paccola CC, Neves FM, de Oliva SU, Miraglia SM (2016) Late reproductive analysis in rat male offspring exposed to nicotine during pregnancy and lactation. Andrology 4:218-231 https://doi.org/10.1111/andr.12100
  36. Zlatkis A, Zak B, Boyle AJ (1953) A new method for the direct determination of serum cholesterol. J Lab Clin Med 41:486-492
  37. Bergmeyer HU (1974) Beta-hydroxysteroid dehydrogenase. In: Bergmeyer HU (ed.) Methods Enzy Analy Academic Press, New York, pp 447-489
  38. Misra HP, Fridovich I (1972) The role of superoxide anion in the autoxidation of epinephrine and a simple assay for superoxide dismutase. J Biol Chem 247:3170-3175 https://doi.org/10.1016/S0021-9258(19)45228-9
  39. Chance B, Maehly AC (1955) Assay of catalase and peroxidises. Methods Enzymol 2:764-775 https://doi.org/10.1016/S0076-6879(55)02300-8
  40. Flohe L, Gunzler WA (1984) Assays of glutathione peroxidise. Methods Enzymol 105:114-120 https://doi.org/10.1016/S0076-6879(84)05015-1
  41. Carlberg I, Mannervik B (1985) Glutathione reductase assay. Methods Enzymol 113:484-495 https://doi.org/10.1016/S0076-6879(85)13062-4
  42. Beutler E (1975) The preparation of red cells for assay. In: Beutler E (ed) Red cell metabolism: a manual of biochemical methods. Grune and Straton Company, New York, pp 8-18
  43. Ohkawa H, Ohishi N, Yagi K (1979) Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem 95:351-358 https://doi.org/10.1016/0003-2697(79)90738-3
  44. Levine RL, Garland D, Oliver CN, Amici A, Climent I, Lenz AG, Ahn BW, Shaltiel S, Stadtman ER (1990) Determination of carbonyl content in oxidatively modified proteins. Methods Enzymol 186:464-478 https://doi.org/10.1016/0076-6879(90)86141-H
  45. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193:265-275 https://doi.org/10.1016/S0021-9258(19)52451-6
  46. Cid C, Alvarez-Cermeno JC, Regidor I, Plaza J, Salinas M, Alcazar A (2003) Caspase inhibitors protect against neuronal apoptosis induced by cerebrospinal fluid from multiple sclerosis patients. J Neuroimmunol 136:119-124 https://doi.org/10.1016/S0165-5728(02)00467-8
  47. Brancraft JD, Stevens A (1982) Theory and practice of histological techniques, 2nd edn. Chruchill Livingstone, New York
  48. Turner TT, Lysiak JJ (2008) Oxidative stress: a common factor in testicular dysfunction. J Androl 29:488-498 https://doi.org/10.2164/jandrol.108.005132
  49. Stocco DM, Clark BJ (2000) Role of the steroidogenic acute regulatory protein (StAR) in steroidogenesis. Biochem Pharmacol 51:197-205 https://doi.org/10.1016/0006-2952(95)02093-4
  50. Clair E, Mesnage R, Travert C, Seralini GE (2012) A glyphosate-based herbicide induces necrosis and apoptosis in mature rat testicular cells in vitro and testosterone decrease at lower levels. Toxicol In Vitro 26:269-279 https://doi.org/10.1016/j.tiv.2011.12.009
  51. Victor-Costa AB, Bandeira SM, Oliveira AG, Mahecha GA, Oliveira CA (2010) Changes in testicular morphology and steroidogenesis in adult rats exposed to atrazine. Reprod Toxicol 29:323-331 https://doi.org/10.1016/j.reprotox.2009.12.006
  52. Knez J (2013) Endocrine-disrupting chemicals and male reproductive health. Reprod Biomed Online 26:440-448 https://doi.org/10.1016/j.rbmo.2013.02.005
  53. Mehrpour O, Karrari P, Zamani N, Tsatsakis AM, Abdollahi M (2014) Occupational exposure to pesticides and consequences on male semen and fertility: a review. Toxicol Lett 230:146-156 https://doi.org/10.1016/j.toxlet.2014.01.029
  54. Espinoza Navarro O, Bustos Obregon E (2014) Effect of Malathion on cellularity and sperm differentiation in testis and eididymis of adult rats. Int J Morphol 32:119-124 https://doi.org/10.4067/S0717-95022014000100020
  55. Nakai M, Hess RA, Moore BJ, Guttroff RF, Strader LF, Linder RE (1992) Acute and long-term effects of a single dose of the fungicide carbenduim (metyl-2-benzemidazole carbamate) on the male reproductive system in the rat. J Androl 13:507-518
  56. Hess RA, Gist DH, Bunick D, Lubahn DB, Farrell A, Bahr J, Cooke PS, Greene GL (1997) Estrogen receptor (alpha and beta) expression in the excurrent ducts of the adult male rat reproductive tract. J Androl 18:602-611
  57. Spirhanzlova P, De Groef B, Nicholson FE, Grommen SVH, Marras G, Sebillot A, Demeneix BA, Pallud-Mothre S, Lemkine GF, Tindall AJ, Du Pasquier D (2017) Using short-term bioassays to evaluate the endocrine disrupting capacity of the pesticides linuron and fenoxycarb. Comp Biochem Physiol C Toxicol Pharmacol 200:52-58 https://doi.org/10.1016/j.cbpc.2017.06.006
  58. U S EPA (1995) Reregistration Eligibility Decision (RED): Linuron. US Environmental Protection Agency, Office of Prevention, Pesticides and Toxic Substances, Office of Pesticide. Programs US Government Printing Office, Washington DC, EPA 738-R-95-003
  59. Pratap Reddy K, Girish BP, Sreenivasula Reddy P (2014) Reproductive and paternal mediated developmental toxicity of Benzo(a)Pyrene in adult male Wistar rats. Toxicol Res (Camb). https://doi.org/10.1039/c4tx00121d
  60. Hamdy BG, Nabil T, Saad NN, Abdelwahab M, Mohamed L (2016) Protective role of alpha lipoic acid against the deleterious effects of both natural and artificial sweetener (Sucrose and Aspartame) in albino rats. Alex J Vet Sci 49:105-115
  61. Lebda M, Gad S, Gaafar H (2014) Effects of lipoic acid on acrylamide induced testicular damage. Mater Soc Med 26:208-212 https://doi.org/10.5455/msm.2014.26.208-212
  62. Holmquist L, Stuchbury G, Berbaum K, Muscat S, Young S, Hager K, Engel J, Munch G (2007) Lipoic acid as a novel treatment for alzheimer's disease and related dementias. Pharmacol Ther 113:154-164 https://doi.org/10.1016/j.pharmthera.2006.07.001
  63. Biewenga GP, Haenen GR, Bast A (1997) The pharmacology of the antioxidant lipoic acid. Gen Pharmacol 29:315-331 https://doi.org/10.1016/S0306-3623(96)00474-0
  64. Shila S, Kokilavani V, Subathra M, Panneerselvam C (2005) Brain regional responses in antioxidant system to alpha-lipoic acid in arsenic intoxicated rat. Toxicology 210:25-36 https://doi.org/10.1016/j.tox.2005.01.003
  65. McIlwain DR, Berger T, Mak TW (2013) Caspase functions in cell death and disease. Cold Spring Harb Perspect Biol 5:a008656 https://doi.org/10.1101/cshperspect.a008656
  66. Santos PS, Campelo LM, Freitas CM, Saldanha GB, Freitas RM (2011) Lipoic acid effects on glutamate and taurine concentrations in rat hippocampus after pilocarpine-induced seizures. Arq Neuropsiquiatr 69:360-364 https://doi.org/10.1590/S0004-282X2011000300018
  67. Astiz M, de Alaniz MJT, Marra CA (2012) The oxidative damage and inflammation caused by pesticides are reverted by lipoic acid in rat brain. Neurochem Int 61:1231-1241 https://doi.org/10.1016/j.neuint.2012.09.003