In vitro protection of human lymphocytes from toxic effects of chlorpyrifos by selenium-enriched medicines

Document Type : Original Article


1 Pharmaceutical Sciences Research Center, Tehran University of Medical Sciences, Tehran, Iran. Department of Tissue Engineering, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran

2 Toxicology and Poisoning Research Center and Department of Toxicology and Pharmacology, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran

3 Pharmaceutical Sciences Research Center, Tehran University of Medical Sciences, Tehran, Iran

4 Pharmaceutical Sciences Research Center, Tehran University of Medical Sciences, Tehran, Iran. Toxicology and Poisoning Research Center and Department of Toxicology and Pharmacology, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran


Objective(s):  Chlorpyrifos (CP) is a broad-spectrum organophosphorus pesticide used extensively in agricultural and domestic pest control, accounting for 50% of the global insecticidal use. In the present study, protective effects of two selenium-enriched strong antioxidative medicines IMOD and Angipars were examined in human lymphocytes treated with CP in vitro.
Materials and Methods: Isolated lymphocytes were exposed to 12 µg/ml CP either alone or in combination with effective doses (ED50) of IMOD (0.2 µg/ml) and Angipars (1 µg/ml). After 3 days incubation, the viability and oxidative stress markers including cellular lipid peroxidation (LPO), myeloperoxidase (MPO), total thiol molecules (TTM), and total antioxidant power (TAP) were evaluated. Also, the levels of tumor necrosis factor-α (TNF-α), as inflammatory index along with acetylcholinesterase (AChE) activity and cell apoptosis were assessed by flow cytometry.
Results: Results indicated that effective doses of IMOD and Angipars reduced CP-exposed lymphocyte mortality rate along with oxidative stress. Both agents restored CP-induced elevation of TNF-α and protected the lymphocytes from CP-induced apoptosis and necrosis.
Conclusion: Overall, results confirm that IMOD and Angipars reduce the toxic effects associated with CP through free radical scavenging and protection from apoptosis and necrosis.


1.Casida JE, Quistad GB. Organophosphate toxicology: safety aspects of nonacetylcholinesterase secondary targets. Chem Res Toxicol 2004; 17:983-998.
2.Tian Y, Ishikawa H, Yamaguchi T, Yamauchi T, Yokoyama K. Teratogenicity and developmental toxicity of chlorpyrifos: Maternal exposure during organogenesis in mice. Reprod Toxicol 2005; 20:267-270.
3.Casida JE. Pest toxicology: the primary mechanisms of pesticide action. Chem Res Toxicol 2009; 22:609-619.
4.Moser VC. Dose-response and time-course of neurobehavioral changes following oral chlorpyrifos in rats of different ages. Neurotoxicol Teratol 2000; 22:713-723.
5.Soltaninejad K, Abdollahi M. Current opinion on the science of organophosphate pesticides and toxic stress: a systematic review. Med Sci Monit 2009; 15(3):RA75-90.
6.Mostafalou S, Abdollahi M. Pesticides and human chronic diseases: evidences, mechanisms, and perspectives. Toxicol Appl Pharmacol 2013; 268(2):157-77.
7.Gupta SC, Mishra M, Sharma A, Deepak Balaji TG, Kumar R, Mishra R K, et al. Chlorpyrifos induces apoptosis and DNA damage in Drosophila through generation of reactive oxygen species. Ecotoxicol Environ Saf 2010; 73:1415-1423.
8.Ishii N, Senoo-Matsuda N, Miyake K, Yasuda K, Ishii T, Hartman PS, et al. Coenzyme Q10 can prolong C. elegans lifespan by lowering oxidative stress. Mech Ageing Dev 2004; 125:41-46.
9.Hundekari IA, Suryakar AN, Rathi DB. Acute organo-phosphorus pesticide poisoning in North Karnataka, India: oxidative damage, haemoglobin level and total leukocyte. Afr Health Sci 2013; 13:129-136.
10.Ambali SF, Abubakar AT, Shittu M, Yaqub LS, Anafi SB, Abdullahi A. Chlorpyrifos-induced alteration of hematological parameters in Wistar rats: ameliorative effect of zinc. Res J Environ Toxicol 2010; 4:55-66.
11.Gultekin F, Delibas N, Yasar S, Kilinc I. 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 of Toxicol 2001; 75:88-96.
12.Ambali S, Akanbi D, Igbokwe N, Shittu M, Kawu M, Ayo J. Evaluation of subchronic chlorpyrifos poisoning on hematological and serum biochemical changes in mice and protective effect of vitamin C. J Toxicol Sci 2007; 32:111-120.
13.Ambali SF, Ayo JO, Ojo SA, Esievo KA. Ameliorative effect of vitamin C on chlorpyrifos-induced increased erythrocyte fragility in Wistar rats. Hum Exp Toxicol 2010; 30:19-24.
14.Ambali SF, Abubakar AT, Shittu M, Yaqub LS, Kobo PI, Giwa A. Zinc ameliorates chlorpyrifos-induced increased erythrocyte fragility and lipoperoxidative changes in Wistar rats. New York Sci J 2010; 3:117-122.
15.Ambali SF, Ayo JO, Esievo KA, Ojo SA. Hemotoxicity induced by chronic chlorpyrifos exposure in Wistar rats: mitigating effect of vitamin C. Vet Med Int. 2011; 2011:945439.
16.Mohammadirad A, Khorram-Khorshid HR, Gharibdoost F, Abdollahi M. Setarud (IMOD™) as a multiherbal drug with promising benefits in animal and human studies: a comprehensive review of biochemical and cellular evidences. Asian J Anim Vet Adv 2011; 6:1185-1192.
17.Hasani-Ranjbar S, Larijani B, Abdollahi M. A systematic review of the potential herbal sources of future drugs effective in oxidant-related diseases. Inflamm Allergy Drug Targets 2009; 8:2-10.
18.Mohseni-Salehi-Monfared SS, Habibollahzadeh E, Sadeghi H, Baeeri M, Abdollahi M. Efficacy of Setarud (IMODTM), a novel electromagnetically-treated multi-herbal compound, in mouse immunogenic type-1 diabetes. Arch Med Sci 2010; 6:663-669.
19.Larijani B, Salimi M, Pourkhalili N, Mohammadirad A, Baeeri M, Nili-Ahmadabadi A, et al. Positive response of isolated rat pancreatic islets to IMOD; hopes for better transplant outcome and graft function. Asian J Anim Vet Adv 2011; 6:1019-1025.
20.Baghaei A, Esmaily H, Abdolghaffari AH, Baeeri M, Gharibdoost F , Abdollahi M. Efficacy of setarud (IMOD), a novel drug with potent anti-toxic stress potential in rat inflammatory bowel disease and comparison with dexamethasone and infliximab. Indian J Biochem Biophys 2010; 47:219-226.
21.Mahmoodpoor A, Eslami K, Mojtahedzadeh M, Najafi A, Ahmadi A, Dehnadi-Moghadam A, et al. Examination of setarud (IMODTM) in the management of patients with severe sepsis. Daru 2010; 18:23-28.
22.Mousavi-Jazi M, Aslroosta H, Moayer A R, Baeeri M, Abdollahi M. Effects of Angipars on oxidative inflammatory indices in a murine model of periodontitis. Daru 2010; 18:260-264.
23.Larijani B, Hasani Ranjbar S. Overview of diabetic foot; novel treatments in diabetic foot ulcer. Daru 2008; 16:1-6.
24.Plesca-Manea L, Parvu A E, Parvu M, Taamas M, Buia R, Puia M. Effects of Melilotus officinalis on acute inflammation. Phytother Res 2002; 16:316-319.
25.Ghanbari S, Yonessi M, Mohammadirad A, Gholami M, Baeeri M, Khorram-Khorshid HR, et al. Effects of IMOD™ and Angipars™ on mouse D-galactose-induced model of aging. Daru 2012; 20:68-75.
26.Shankarjit S, Aruna B. Biotherapeutic Potential of Ziziphus Mauritiana (Lamk.) Extract against chlorpyrifos induced oxidative stress (an in-vitro study). Biochem Mol Biol 2013; 1:58-62.
27.Caughlan A, Newhouse K, Namgung U, Xia Z. Chlorpyrifos induces apoptosis in rat cortical neurons that is regulated by a balance between p38 and ERK/JNK MAP kinases. Toxicol Sci 2004; 78:125-134.
28.Sharma V, Singh M. Attenuation of N-nitrosodimethylamine induced hepatotoxicity by Operculina turpethum in Swiss Albino mice. Iran J Basic Med Sci 2014; 17:73-80.
29.Ghazanfari G, Minaie B, Yasa N, Nakhai LA, Mohammadirad A, Nikfar S, et al. Biochemical and histopathological evidences for beneficial effects of Satureja khuzestanica jamzad essential oil on the mouse model of inflammatory bowel diseases. Toxicol Mech Meth 2006; 16:365-372.
30.Aliabadi A, Foroumadi A, Mohammadi-Farani A, Garmsiri Mahvar M. Synthesis and evaluation of anti-acetylcholinesterase activity of 2-(2-(4-(2-Oxo-2-phenylethyl) piperazin-1-yl) ethyl) isoindoline-1,3-dione derivatives with potential anti-Alzheimer effects. Iran J Basic Med Sci 2013; 16:1049-1054.
31.Kumar A, Malik F, Bhushan S, Sethi VK, Shahi AK, Kaur J, et al. An essential oil and its major constituent isointermedeol induce apoptosis by increased expression of mitochondrial cytochrome c and apical death receptors in human leukaemia HL-60 cells. Chem Biol Interact 2008; 171:332-347.
32.Elelaimy IA, Ibrahim HM, Abdel Ghaffar FR, Alawthan YS. Evaluation of sub-chronic chlorpyrifos poisoning on immunological and biochemical changes in rats and protective effect of eugenol. J Appl Pharm Sci 2012; 2:51-61.
33.Blakley BR, Yole MJ, Brousseau P, Boermans H, Fournier M. Effect of chlorpyrifos on immune function in rats. Vet Hum Toxicol 1999; 41:140-144.
34.Thrasher JD, Heuser G, Broughton A. Immunological abnormalities in humans chronically exposed to chlorpyrifos. Arch Environ Health 2002; 57:181-187.
35.Węgorek P, Mrowczyński M, Zamojska J. Resistance of pollen beetle (Meligethes aeneus F.) to selected active substances of insecticides in Poland. J Plant Protect Res 2009; 49:199-127.
36.Gultekin F, Ozturk M, Akdogan M. The effect of organophosphate insecticide chlorpyrifos-ethyl on lipid peroxidation and antioxidant enzymes (in vitro). Arch Toxicol 2000; 74:553-538.
37.Kehrer JP. Free radicals as mediators of tissue injury and disease. Crit Rev Toxicol 1993; 23:21-48.
38.Datta C, Gupta J, Sarkar A, Senqupta D. Effects of organophosphorus insecticide phosphomidon on antioxidant defence components of human erythrocyte and plasma. Indian J Exp Biol 1992; 30:65-67.
39.Uzun FG, Demir F, Kalender S, Bas H, Kalender Y. Protective effect of catechin and quercetin on chlorpyrifos-induced lung toxicity in male rats. Food Chem Toxicol 2010; 48:1714-1720.
40.Zhang R, Brennan ML, Shen Z, MacPherson JC, Schmitt D, Molenda CE, et al. Myeloperoxidase functions as a major enzymatic catalyst for initiation of lipid peroxidation at sites of inflammation. J Biol Chem 2002; 277:46116-4622.
41.Mehri A, Hasani-Ranjbar S, Larijani B, Abdollahi M. A systematic review of efficacy and safety of Urtica dioica in the treatment of diabetes. Int J Pharmacol 2001; 7:161-170.
42.Köhrle J, Jakob F, Contempré B, Dumont JE. Selenium, the thyroid, and the endocrine system. Endocr Rev 2005; 26:944-984.
43.Hassani-Ranjbar S, Nayebi N, Larijani B, Abdollahi M. A systematic review of the efficacy and safety of Teucrium species; from anti-oxidant to anti-diabetic effects. Int J Pharmacol 2010; 6:315-325.
44.Miroliaee AE, Esmaily H, Vaziri-Bami A, Baeeri M, Shahverdi AR, Abdollahi M. Amelioration of experimental colitis by a novel nanoselenium–silymarin mixture. Toxicol Mech Methods 2011; 21:200-208.
45.Tabatabaei-Malazy O, Larijani B, Abdollahi M. A novel management of diabetes by means of strong antioxidants’ combination. J Med Hypotheses Ideas 2013; 7:25-30.
46.Nakadai A, Li Q, Kawada T. Chlorpyrifos induces apoptosis in human monocyte cell line U937. Toxicology 2006; 224:202-209.
47.Greenlee AR, Ellis TM, Berg RL. Low-dose agrochemicals and lawn-care pesticides induce developmental toxicity in murine preimplantation embryos. Environ Health Perspect 2004; 112:703-709.
48.Thrasher JD, Madison R, Broughton A. Immunologic abnormalities in humans exposed to chlorpyrifos: preliminary observations. Arch Environ Health 1993; 48:89-93.
49.Corcoran GB, Fix L, Jones DP, Moslen MT, Nicotera P, Oberhammer FA, et al. Apoptosis: molecular control point in toxicity, Toxicol Appl Pharmacol 1994; 128:169-181.
50.Karami-Mohajeri S, Abdollahi M. Mitochondrial dysfunction and organophosphorus compounds. Toxicol Appl Pharmacol 2013; 270(1):39-44.
51.Crumpton TL, Seidler FJ, Slotkin TA, Developmental neurotoxicity of chlorpyrifos in vivo and in vitro: effects on nuclear transcription factors involved in cell replication and differentiation. Brain Res 2000; 857:87-98.
52.Song X, Seidler FJ, Saleh JL, Zhang J, Padilla S, Slotkin TA. Cellular mechanisms for developmental toxicity of chlorpyrifos: targeting the adenylyl cyclase signaling cascade. Toxicol Appl Pharmacol 1997; 145:158-174.
53.Palmer HJ, Paulson KE. Reactive oxygen species and antioxidants in signal transduction and gene expression. Nutr Rev 1997; 55:353-361.