Antioxidant attenuation of ROS-involved cytotoxicity induced by Paraquat on HL-60 cells
Wen-Hua Zhang, Yang Yang, Chang-Jun Lin, Qin Wang
Lanzhou University.
DOI: 10.4236/health.2010.23036   PDF    HTML     6,668 Downloads   12,051 Views   Citations


The cytotoxic effects of Paraquat, an herbicide refractory to treatment after intentional or accidental contact, were investigated on the human leukemia HL-60 cells. With the establishment of Paraquat injury model of HL-60 cells, trypan blue exclusion assaying was performed to have determined the effects of Paraquat-induced cytotoxicity on HL-60 cells in a concentration- dependent manner. Upon treatment with various concentrations of Paraquat, pronounced increase on the levels of intracellular production of O2?- and H2O2 was detected with employment of fluorescent probes. Indicative of the oxidative stress, levels of MDA and T-AOC were quantitated to have determined the causal role for Paraquat in subjecting HL-60 cells to oxidative damage. Based on this finding, effects of antioxidant enzymes including GSH, NAC, CAT and SOD on attenuating the Paraquat-induced oxidative damage on HL-60 cells were examined, aiming to identify the most effective antioxidant enzyme for alleviating the cytotoxicity induced by Paraquat. In conjunction with the determination of cytotoxicity exerted by all the antioxidant enzymes on HL-60 cells, GSH-with its least inherent cytotoxicity on HL-60 cells-was identified as a promising candidate ingredient for extenuating the Paraquat-induced cytotoxicity.

Share and Cite:

Zhang, W. , Yang, Y. , Lin, C. and Wang, Q. (2010) Antioxidant attenuation of ROS-involved cytotoxicity induced by Paraquat on HL-60 cells. Health, 2, 253-261. doi: 10.4236/health.2010.23036.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] Clark, D.G., McElligott, T.F. and Hurst, E.W. (1966) The toxicity of Paraquat. British Journal of Industrial Medicine, 23, 126-132.
[2] Bullivant, C.M. (1966) Accidental poisoning by Paraquat: Report of two cases in man. British Medical Journal, 1, 1272-1273.
[3] Somayajulu-Nitu, M., Sandhu, J.K., Cohen, J., Sikorska, M., Sridhar, T.S., Matei, A., Borowy-Borowski, H. and Pandey, S. (2009) Paraquat induces oxidative stress, neuronal loss in substantia nigra region and Parkinsonism in adult rats: Neuroprotection and amelioration of symptoms by water-soluble formulation of Coenzyme Q10. BMC Neuroscience, 10, 88.
[4] Roberts, T.R., Dyson, J.S. and Lane, M.C. (2002) Deactivation of the biological activity of Paraquat in the soil environment: A review of long-term environmental fate. Journal of Agricultural and Food Chemistry, 50, 3623- 3631.
[5] Onyon, L.J. and Volans, G.N. (1987) The epidemiology and prevention of Paraquat poisoning. Hum Toxicol, 6, 19-29.
[6] Bismuth, C., Garnier, R., Baud, F.J., Muszynski, J. and Keyes, C. (1990) Paraquat poisoning: An overview of the current status. Drug Safety, 5, 243-251.
[7] Botella de Maglia, J. and Belenguer Tarin, J.E. (2000) Paraquat poisoning: A study of 29 cases and evaluation of the effectiveness of the “Caribbean scheme”. Medicina Clinica (Barc), 115, 530-533.
[8] Luo, Z.Y. (2007) Treatment 16 cases of acute Paraquat poisoning. Chinese Journal of Labour Hygiene and Occupational Diseases, 25, 330.
[9] Zhao, F.L., Wang, J. and Guo, W. (2009) Clinical observation on 22 cases of acute Paraquat poisoning. Chinese Journal of Labour Hygiene and Occupational Diseases, 27, 56-57.
[10] Rose, M.S., Lock, E.A., Smith, L.L. and Wyatt, I. (1976) Paraquat accumulation: Tissue and species specificity. Biochemical Pharmacology, 25, 419-423.
[11] Rose, M.S., Smith, L.L. and Wyatt, I. (1974) Evidence for energy-dependent accumulation of Paraquat into rat lung. Nature, 252, 314-315.
[12] Melchiorri, D., Del Duca, C., Piccirilli, S., Trombetta, G., Bagetta, G. and Nistico, G. (1998) Intrahippocampal injection of Paraquat produces apoptotic cell death which is prevented by the lazaroid U74389G, in rats. Life Sciences, 62, 1927-1932.
[13] Barone, S., Das, K.P., Lassiter, T.L. and White, L.D. (2000) Vulnerable processes of nervous system development: A review of markers and methods. Neurotoxicology, 21, 15-36.
[14] Gonzalez-Polo, R.A., Niso-Santano, M., Ortiz-Ortiz, M.A., Gomez-Martin, A., Moran, J.M., Garcia-Rubio, L., Francisco-Morcillo, J., Zaragoza, C., Soler, G. and Fuentes, J.M. (2007) Relationship between autophagy and apoptotic cell death in human neuroblastoma cells treated with Paraquat: Could autophagy be a “brake” in Paraquat-induced apoptotic death? Autophagy, 3, 366-367.
[15] Chen, P., Chen, Z., Li, A., Lou, X.C., Wu, X.K., Zhao, C.J., Wang, S.L. and Liang, L.P. (2008) Catalytic metalloporphyrin protects against Paraquat neurotoxicity in vivo. Biomedical and Environmental Sciences, 21, 233-238.
[16] Drechsel, D.A. and Patel, M. (2008) Role of reactive oxygen species in the neurotoxicity of environmental agents implicated in Parkinson’s disease. Free Radical Biology & Medicine, 44, 1873-1886.
[17] Kuo, M.L. and Lin, J.K. (1993) The genotoxicity of the waste water discharged from Paraquat manufacturing and its pyridyl components. Mutation Research, 300, 223-229.
[18] Takeyama, N., Tanaka, T., Yabuki, T. and Nakatani, T. (2004) The involvement of p53 in Paraquat-induced apoptosis in human lung epithelial-like cells. International Journal of Toxicology, 23, 33-40.
[19] Haddad, J.J. (2004) Redox and oxidant-mediated regulation of apoptosis signaling pathways: Immuno-pharmaco-redox conception of oxidative siege versus cell death commitment. International Immunopharmacology, 4, 475-493.
[20] Farrington, J.A., Ebert, M., Land, E.J. and Fletcher, K. (1973) Bipyridylium quaternary salts and related compounds V. Pulse radiolysis studies of the reaction of Paraquat radical with oxygen: Implications for the mode of action of bipyridyl herbicides. Biochimica et Biophysica Acta, 314, 372-381.
[21] Tsukamoto, M., Tampo, Y., Sawada, M. and Yonaha, M. (2000) Paraquat-induced membrane dysfunction in pulmonary microvascular endothelial cells. Pharmacol Toxicol, 86, 102-109.
[22] Tsukamoto, M., Tampo, Y., Sawada, M. and Yonaha, M. (2002) Paraquat-induced oxidative stress and dysfunction of the glutathione redox cycle in pulmonary microvascular endothelial cells. Toxicology and Applied Pharmacology, 178, 82-92.
[23] Zang, L.Y., van Kuijk, F.J. and Misra, H.P. (1995) EPR studies of spin-trapped free radicals in Paraquat-treated lung microsomes. Biochemistry & Molecular Biology International, 37, 255-262.
[24] Yang, W.L. and Sun, A.Y. (1998) Paraquat-induced cell death in PC12 cells. Neurochemical Research, 23, 1387- 1394.
[25] Solaini, G. and Harris, D.A. (2005) Biochemical dysfunction in heart mitochondria exposed to ischaemia and reperfusion. Biochemical Journal, 390, 377-394.
[26] Blomgren, K. and Hagberg, H. (2006) Free radicals, mitochondria, and hypoxia-ischemia in the developing brain. Free Radical Biology & Medicine, 40, 388-397.
[27] Dringen, R. (2000) Metabolism and functions of glutathione in brain. Progress in Neurobiology, 62, 649-671.
[28] Dringen, R. (2005) Oxidative and antioxidative potential of brain microglial cells. Antioxid Redox Signal, 7, 1223-1233.
[29] Floyd, R.A. and Hensley, K. (2002) Oxidative stress in brain aging: Implications for therapeutics of neurodegenerative diseases. Neurobiology of Aging, 23, 795-807.
[30] Frasca, S., Couvreur, P., Seiller, M., Pareau, D., Lacour, B., Stambouli, M. and Grossiord, J.L. (2009) Paraquat detoxication with multiple emulsions. International Journal of Pharmaceutics, 380, 142-146.
[31] Hays, P.G. (1995) VIP interview: Patrick G. Hays. Interview by Maria R. Traska. Managed Care Quarterly, 3, 1-6.
[32] LeBel, C.P., Ischiropoulos, H. and Bondy, S.C. (1992) Evaluation of the probe 2’, 7’-dichlorofluorescin as an indicator of reactive oxygen species formation and oxidative stress. Chemical Research in Toxicology, 5, 227-231.
[33] Benov, L., Sztejnberg, L. and Fridovich, I. (1998) Critical evaluation of the use of hydroethidine as a measure of superoxide anion radical. Free Radical Biology & Medicine, 25, 826-831.
[34] Oubrahim, H., Stadtman, E.R. and Chock, P.B. (2001) Mitochondria play no roles in Mn(II)-induced apoptosis in HeLa cells. Proceedings of the National Academy of Sciences USA, 98, 9505-9510.
[35] Lee, C.S., Han, E.S. and Lee, W.B. (2003) Antioxidant effect of phenelzine on MPP+-induced cell viability loss in differentiated PC12 cells. Neurochemical Research, 28, 1833-1841.
[36] Miyata, K., Rahman, M., Shokoji, T., Nagai, Y., Zhang, G.X., Sun, G.P., Kimura, S., Yukimura, T., Kiyomoto, H., Kohno, M., Abe, Y. and Nishiyama, A. (2005) Aldosterone stimulates reactive oxygen species production through activation of NADPH oxidase in rat mesangial cells. Journal of American Society of Nephrology, 16, 2906-2912.
[37] Ohkawa, H., Ohishi, N. and Yagi, K. (1979) Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Analytical Biochemistry, 95, 351-358.
[38] Matthew, H., Logan, A., Woodruff, M.F. and Heard, B. (1968) Paraquat poisoning-lung transplantation. British Medical Journal, 3, 759-763.
[39] McDonagh, B.J. and Martin, J. (1970) Paraquat poisoning in children. Archives of Disease in Childhood, 45, 425-427.
[40] Wesseling, C., Hogstedt, C., Picado, A. and Johansson, L. (1997) Unintentional fatal Paraquat poisonings among agricultural workers in Costa Rica: Report of 15 cases. American Journal of Industrial Medicine, 32, 433-441.
[41] Hantson, P. (2008) Paraquat poisoning. Rev Prat, 58, 866-867.
[42] Zheng, X.B. and Sun, X.L. (2008) Treatment of Paraquat poisoning. Chinese Journal of Labour Hygiene and Occupational Diseases, 26, 635-637.
[43] Ikebuchi, J. (1987) Evaluation of Paraquat concentrations in Paraquat poisoning. Archives of Toxicology, 60, 304- 310.
[44] Senarathna, L., Eddleston, M., Wilks, M.F., Woollen, B.H., Tomenson, J.A., Roberts, D.M. and Buckley, N.A. (2009) Prediction of outcome after Paraquat poisoning by measurement of the plasma Paraquat concentration. Oxford Journal of Medicine, 102, 251-259.
[45] Lee, K.H., Gil, H.W., Kim, Y.T., Yang, J.O., Lee, E.Y. and Hong, S.Y. (2009) Marked recovery from Paraquat- induced lung injury during long-term follow-up. Korean Journal of Internal Medicine, 24, 95-100.
[46] Fabiani, R., Fuccelli, R., Pieravanti, F., De Bartolomeo, A. and Morozzi, G. (2009) Production of hydrogen peroxide is responsible for the induction of apoptosis by hydroxytyrosol on HL60 cells. Molecular Nutrition & Food Research, 53, 887-896.
[47] Schmuck, G., Rohrdanz, E., Tran-Thi, Q.H., Kahl, R. and Schluter, G. (2002) Oxidative stress in rat cortical neurons and astrocytes induced by Paraquat in vitro. Neurotoxicity Research, 4, 1-13.
[48] Mates, J.M., Segura, J.M., Perez-Gomez, C., Rosado, R., Olalla, L., Blanca, M. and Sanchez-Jimenez, F.M. (1999) Antioxidant enzymatic activities in human blood cells after an allergic reaction to pollen or house dust mite. Blood Cells, Molecules, and Diseases, 25, 103-109.
[49] Mates, J.M., Perez-Gomez, C. and Blanca, M. (2000) Chemical and biological activity of free radical ‘scavengers’ in allergic diseases. Clinica Chimica Acta, 296, 1-15.

Copyright © 2023 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.