Advertisement

Protective role of citric acid against oxidative stress induced by heavy metals in Caenorhabditis elegans

  • Shaojuan SongEmail author
  • Yan Han
  • Yun Zhang
  • Honglian Ma
  • Lei Zhang
  • Jing Huo
  • Peisheng Wang
  • Mengrui Liang
  • Ming Gao
Research Article

Abstract

The adverse effects of heavy metals, such as cadmium, zinc, and copper, occur due to the generation of reactive oxygen species (ROS). The use of Caenorhabditis elegans for the purposes of conservation and biomonitoring is of great interest. In the present study, ROS, malondialdehyde (MDA), and citric acid levels and superoxide dismutase (SOD) and glutathione peroxidase (GPx) activities in a model organism were tested to study toxicity. C. elegans was exposed to three different concentrations of cadmium (CdCl2, 5, 10, 50 μM), zinc (ZnSO4, 10, 100, 500 μM), and copper (CuSO4, 10, 100, 500 μM) for 3 days. ROS levels increased by 1.3- to 2.1-fold with increasing metal concentrations. The MDA content increased by approximately 7-, 5-, 2-fold after exposure to high concentrations of cadmium, zinc, and copper, respectively. Furthermore, the citric acid content increased by approximately 3-fold in the cadmium (Cd, 5 μM), zinc (Zn, 10 μM), and copper (Cu, 100 μM) treatment groups compared to that in untreated C. elegans. Therefore, citric acid may play an important role in heavy metal detoxification. Excess citric acid also slightly increased the LC50 by 1.3- to 2.0-fold, basic movements by 1.0- to 1.5-fold, decreased the ROS content by 2.4- to 2.1-fold, the MDA content by 4- to 2-fold, the SOD activity by 9- to 3-fold, the GPx activity by 4.0- to 3.0-fold, and the mRNA expression levels of GPxs by 3.2- to 1.8-fold after metals treatment. And it is most significantly in the alleviation of citric acid to cadmium. This study not only provides information to further understand the effects of heavy metal exposure on ROS, MDA, GPx, SOD, and citric acid in worms but also indicates that supplemental citric acid can protect animals from heavy metal stress and has broad application prospects in decreasing oxidative damage caused by heavy metals.

Keywords

Caenorhabditis elegans ROS MDA Citric acid Antioxidative enzymes Heavy metals 

Notes

Acknowledgements

The authors thank WormBase and WormBook.

Funding information

This work was supported by the Outstanding Graduate Science Innovation Foundation of Shanxi Province (Grant no. 021852901008), the Science and Technology Innovation Team Project of Changzhi Medical College (Grant no. CX201510), and an Innovation and Entrepreneurship Training Program for college students in Shanxi (Grant nos. 2016306, 2016309, 2016314).

References

  1. Acosta C, Barat JM, Martínezmáñez R, Sancenón F, Llopis S, González N, Genoves S, Ramon D, Martorell P (2018) Toxicological assessment of mesoporous silica particles in the nematode Caenorhabditis elegans. Environ Res 166:61–70CrossRefGoogle Scholar
  2. Agnello AC, Huguenot D, Hullebusch EDV (2016) Citric acid- and Tween® 80-assisted phytoremediation of a co-contaminated soil: alfalfa (Medicago sativa L.) performance and remediation potential. Environ Sci Pollut Res 2016:1–12Google Scholar
  3. Ahn JM, Eom HJ, Yang XY, Meyer JN, Choi J (2014) Comparative toxicity of silver nanoparticles on oxidative stress and DNA damage in the nematode, Caenorhabditis elegans. Chemosphere. 108:343–352CrossRefGoogle Scholar
  4. Arsenov D, Zupunski M, Borisev M, et al. (2016) Antioxidative response of Salix spp. in citric acid assisted phytoremediation of cadmium. Plant Biology Europe EPSO/FESPB 2016 CongressGoogle Scholar
  5. Auer T, Khoschsorur GA, Rabl H, Iberer F, Petutschnigg B, Wasler A, Tscheliessnigg KH (1995) Detection of lipid peroxidation products by malondialdehyde (MDA-TBA reaction) in organ transplantation. Transplant Proc 27(5):2749–2751Google Scholar
  6. Benáková M, Ahmadi H, Dučaiová Z, Tylová E, Clemens S, Tůma J (2017) Effects of Cd and Zn on physiological and anatomical properties of hydroponically grown Brassica napus plants. Environ Sci Pollut Res Int 241(25):1–12Google Scholar
  7. Brenner S (1974) The genetics of Caenorhabidits elegans. Genetics. 77:71–94Google Scholar
  8. Cheng D, Zhang X, Xu L, Li X, Hou L, Wang C (2017) Protective and prophylactic effects of chlorogenic acid on aluminum-induced acute hepatotoxicity and hematotoxicity in mice. Chem Biol Interact 273:125–132CrossRefGoogle Scholar
  9. Donkin SG, Williams PL (1995) Influence of developmental stage, salts and food presence on various end points using Caenorhabditis elegans for aquatic toxicity testing. Environ Toxicol Chem 14:2139–2147CrossRefGoogle Scholar
  10. Ehsan S, Ali S, Noureen S, Mahmood K, Farid M, Ishaque W, Shakoor MB, Rizwan M (2014) Citric acid assisted phytoremediation of cadmium by Brassica napus L. Ecotoxicol Environ Saf 106:164–172CrossRefGoogle Scholar
  11. Emre I, Kayis T, Coskun M, Dursun O, Cogun HY (2015) Changes in antioxidative enzyme activity, glycogen, lipid, protein, and malondialdehyde content in cadmium-treated Galleria mellonella larvae. Ann Entomol Soc Am 106(3):371–377CrossRefGoogle Scholar
  12. Feng J, Lin Y, Yang Y, Shen Q, Huang J, Wang S, Zhu X, Li Z (2018) Tolerance and bioaccumulation of combined copper, zinc, and cadmium in, Sesuvium portulacastrum. Mar Pollut Bull 131:416–421CrossRefGoogle Scholar
  13. Gandhi S, Santelli J, Mitchell DH, Stiles JW, Sanadi DR (1980) A simple method for maintaining large, aging populations of Caenorhabditis elegans. Mech Ageing Dev 12(2):137–150CrossRefGoogle Scholar
  14. García-Sevillano MA, Contreras-Acuña M, García-Barrera T, Navarro F, Gomez-Ariza JL (2014) Metabolomic study in plasma, liver and kidney of mice exposed to inorganic arsenic based on mass spectrometry. Anal Bioanal Chem 406:1455–1469CrossRefGoogle Scholar
  15. Han Y, Song S, Guo Y, Zhang J, Ma E (2016) Ace-3 plays an important role in phoxim resistance in Caenorhabditis elegans. Ecotoxicology 25(4):835–844Google Scholar
  16. Harrington JM, Boyd WA, Smith MV, Rice JR, Freedman JH, Crumbliss AL (2012) Amelioration of metal-induced toxicity in Caenorhabditis elegans: utility of chelating agents in the bioremediation of metals. Toxicol Sci 129(1):49–56CrossRefGoogle Scholar
  17. Hoogewijs D, Houthoofd K, Matthijssens F, Vandesompele J, Vanfleteren JR (2008) Selection and validation of a set of reliable reference genes for quantitative sod gene expression analysis in C. elegans. BMC Mol Biol 9:1–8CrossRefGoogle Scholar
  18. Hunt PR, Olejnik N, Sprando RL (2012) Toxicity ranking of heavy metals with screening method using adult Caenorhabditis elegans and propidium iodide replicates toxicity ranking in rat. Food Chem Toxicol 50(9):3280–3290CrossRefGoogle Scholar
  19. Kanak EG, Dogan Z, Eroglu A, Atli G, Canli M (2014) Effects of fish size on the response of antioxidant systems of Oreochromis niloticus following metal exposures. Fish Physiol Biochem 2014:1–9Google Scholar
  20. Khanna N, Iii CP, Tatara CP, Williams PL (1997) Tolerance of the nematode Caenorhabditis elegans, to pH, salinity, and hardness in aquatic media. Arch Environ Contam Toxicol 32(1):110–114CrossRefGoogle Scholar
  21. Kumar R, Pradhan A, Khan FA, Lindstrom P, Ragnvaldsson D, Ivarsson P, Olsson PE, Jass J (2015) Comparative analysis of stress induced gene expression in Caenorhabditis elegans following Exposure to environmental and lab reconstituted complex metal mixture. PLoS One 10(7):e0132896CrossRefGoogle Scholar
  22. Kutrowska A, Małecka A, Piechalak A, Masiakowski W, Hanć A, Barałkiewicz D, Andrzejewska B, Zbierska J, Tomaszewska B (2017) Effects of binary metal combinations on zinc, copper, cadmium and lead uptake and distribution in Brassica juncea. J Trace Elem Med Biol 44:32–39CrossRefGoogle Scholar
  23. Lee M, Cho S, Roh K, Chae J, Park JH, Park J, Lee MA, Kim J, Auh CK, Yeom CH, Lee S (2017) Glutathione alleviated peripheral neuropathy in oxaliplatin-treated mice by removing aluminum from dorsal root ganglia[J]. Am J Transl Res 9(3):926–939Google Scholar
  24. Li YJ, Hu PJ, Zhao J, Dong CX (2014) Remediation of cadmium and lead-contaminated agricultural soil by composite washing with chlorides and citric acid. Environ Sci Pollut Res 22(7):5563–5571CrossRefGoogle Scholar
  25. Lin G, Cutright TJ (2015) Effect of citric acid and bacteria on metal uptake in reeds grown in a synthetic acid mine drainage solution. J Environ Manag 150:235–242CrossRefGoogle Scholar
  26. Luo J, Cai L, Qi S, Wu J, Gu XS (2017) A multi-technique phytoremediation approach to purify metals contaminated soil from e-waste recycling site. J Environ Manag 204:17–22CrossRefGoogle Scholar
  27. Mahmud JA, Hasanuzzaman M, Nahar K, Bhuyan MHMB, Fujita M (2018) Insights into citric acid-induced cadmium tolerance and phytoremediation in Brassica juncea L.: coordinated functions of metal chelation, antioxidant defense and glyoxalase systems. Ecotoxicol Environ Saf 147:990–1001CrossRefGoogle Scholar
  28. Mao L, Tang D, Feng HW, Gao Y, Zhou P, Xu LR, Wang LM (2015) Determining soil enzyme activities for the assessment of fungi and citric acid-assisted phytoextraction under cadmium and lead contamination. Environ Sci Pollut Res 2015:1–10Google Scholar
  29. Misra HP, Fridovich I (1972) The univalent reduction of oxygen by reduced flavins and quinones. J Biol Chem 247(1):188–192Google Scholar
  30. Nunes B, Capela RC, Sérgio T, Caldeira C, Goncalves F, Correia AT (2014) Effects of chronic exposure to lead, copper, zinc, and cadmium on biomarkers of the European eel, Anguilla anguilla. Environ Sci Pollut Res 21(8):5689–5700CrossRefGoogle Scholar
  31. Paglia DE, Valentine WN (1967) Studies on the quantitative and qualitative characterization of erythrocyte glutathione peroxidase. J Lab Clin Med 70(1):158–169Google Scholar
  32. Pitcher GW, Sherman CC, Vickeby HB (1936) A method to determine small amounts of citric acid in biological material. J Biol Chem 113(1):235–245Google Scholar
  33. Rao GN, Sadasivudu B, Cotlier E (2017) Studies on glutathione S-transferase, glutathione peroxidase and glutathione reductase in human normal and cataractous lenses. Ophthalmic Res 15(4):173–179CrossRefGoogle Scholar
  34. Read G (2017) The role of gpx-8 in DNA repair of C. elegans in biology. University of Iowa. 29Google Scholar
  35. Saglam D, Atli G, Dogan Z, Baysoy E, Gurler C, Eroglu A, Canli M (2014) Response of the antioxidant system of freshwater fish (Oreochromis niloticus) exposed to metals (Cd, Cu) in differing hardness. Turk J Fish Aquat Sci 14:43–52Google Scholar
  36. Smith PK, Krohn RI, Hermanson GT, Mallia AK, Gartner FH, Provenzano MD, Fujimoto EK, Goeke NM, Olson BJ, Klenk DC (1985) Measurement of protein using bicinchoninic acid. Anal Biochem 150:76–85CrossRefGoogle Scholar
  37. Song S, Guo Y, Yin K, Wu H, Zhang X, Yang M, Ma E (2008) Copper on long-term role of nematode physiological characteristics. Sichuan Journal of Zoology 27:832–834Google Scholar
  38. Song S, Guo Y, Zhang X, Zhang X, Zhang J, Ma E (2014a) Changes to cuticle surface ultrastructure and some biological functions in the nematode Caenorhabditis elegans exposed to excessive copper. Arch Environ Contam Toxicol 66(3):390–399CrossRefGoogle Scholar
  39. Song S, Zhang X, Wu H, Han Y, Zhang J, Ma E, Guo YP (2014b) Molecular basis for antioxidant enzymes in mediating copper detoxification in the nematode Caenorhabditis elegans. PLoS One 9:e107685CrossRefGoogle Scholar
  40. Souid G, Souayed N, Yaktiti F, Maaroufi K (2013) Effect of acute cadmium exposure on metal accumulation and oxidative stress biomarkers of Sparus aurata. Ecotoxicol Environ Saf 89:1–7CrossRefGoogle Scholar
  41. Stylianou M, Björnsdotter MK, Olsson PE, Ericson Jogsten I, Jass J (2019) Distinct transcriptional response of Caenorhabditis elegans to different exposure routes of perfluorooctane sulfonic acid. Environ Res 168:406–413CrossRefGoogle Scholar
  42. Wang DY, Shen LK, Wang Y (2007) The phenotypic and behavioral defects can be transferred from zinc-exposed nematodes to their progeny. Environ Toxicol Pharmacol 24(3):223–230CrossRefGoogle Scholar
  43. Wang JC, Zhu HL, Liu XZ, Liu ZP (2014) N-acetylcysteine protects against cadmium-induced oxidative stress in rat hepatocytes. J Vet Sci 15(4):485–493CrossRefGoogle Scholar
  44. Wu Y, Yang J, Zhou XY, Lei M, Gao D, Qiao PW, Du GD (2015) Risk assessment of heavy metal contamination in farmland soil in Du’an Autonomous County of Guangxi Zhuang Autonomous Region, China. Huan Jing Ke Xue 36(8):2964–2971Google Scholar
  45. Xiong QL, Zhao WJ, Guo XY, Shu TT, Chen FT, Zheng XX, Gong ZN (2015) Dustfall heavy metal pollution during winter in north China. Bull Environ Contam Toxicol 95(4):1–7CrossRefGoogle Scholar
  46. Xu H, Zhang X, Li H, Li C, Huo XJ, Hou LP, Gong Z (2018) Immune response induced by major environmental pollutants through altering neutrophils in zebrafish larvae. Aquat Toxicol 201:99–108CrossRefGoogle Scholar
  47. You X, Xi J, Cao YY, Zhang JF, Luan Y (2017) 4-Bromodiphenyl ether induces germ cell apoptosis by induction of ROS and DNA damage in Caenorhabditis elegans. Toxicol Sci 158:240CrossRefGoogle Scholar
  48. Yu Z-Y, Zhang J, Yin D-Q (2012) Toxic and recovery effects of copper on Caenorhabditis elegans by various food-borne and water-borne pathways. Chemosphere 87(11):1361–1367Google Scholar
  49. Zeitoun-Ghandour S, Charnock JM, Hodson ME, Leszczyszyn OI, Blindauer CA, Stürzenbaum SR (2010) The two Caenorhabditis elegans metallothioneins (CeMT-1 and CeMT-2) discriminate between essential zinc and toxic cadmium. FEBS J 277:2531–2542CrossRefGoogle Scholar
  50. Zhang HC, Shi CY, Sun LQ, Wang F, Chen GW (2015) Toxic effects of ionic liquid l-octyl-3-methylimidazolium bromide on the antioxidant defense system of freshwater planarian, Dugesia japonica. Toxicol Ind Health 2015:1–9Google Scholar
  51. Zhang H, Gao Y, Xiong H (2017) Removal of heavy metals from polluted soil using the citric acid fermentation broth: a promising washing agent. Environ Sci Pollut Res 24(10):1–9Google Scholar
  52. Zhang X, Zhong B, Shafi M, Guo J, Liu C, Guo H, Peng D, Wang Y, Liu D (2018) Effect of EDTA and citric acid on absorption of heavy metals and growth of Moso bamboo. Environ Sci Pollut Res 25(10):1–7Google Scholar
  53. Zhao G, Ye S, Yuan H, Ding X, Wang J (2017) Surface sediment properties and heavy metal pollution assessment in the Pearl River estuary, China. Environ Sci Pollut Res 24:2966CrossRefGoogle Scholar
  54. Zhou Q, Gu YL, Yue X, Mao GC, Wang YF, Su H, Xu J, Shi HB, Zou BB, Zhao JS, Wang RY (2017) Combined toxicity and underlying mechanisms of a mixture of eight heavy metals. Mol Med Rep 15(2):859CrossRefGoogle Scholar

Copyright information

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

Authors and Affiliations

  1. 1.Changzhi Medical CollegeChangzhiChina
  2. 2.School of Life ScienceShanxi UniversityTaiyuanChina

Personalised recommendations