Methanol Leaf Extract of <i>Diospyros chloroxylon</i> Attenuates Chlorpyrifos-Induced Toxicity in Rats


  • G. E. Adeleke
  • O. T. Adedosu
  • O. O. Arinde
  • O. E. Adeyemo
  • W. Bolarinwa
  • E. B. Oyewo


Background and Objective: Chlorpyrifos (CPF) is an Organophosphate insecticide commonly used for household and Agricultural purposes. Despite the several reported toxicity in humans, there has been virtually no alternative effective insecticide. Thus, the amelioration of the toxicities seems the best option in alternative medicine. This study, therefore, examine the effects of methanol extract of Diospyros chloroxylon leaf (MEDCL) on the brain and heart of rats administered with CPF.  Materials and Methods: Twenty-four rats were randomized into four groups of 6 rats each, and administered separately with distilled water (Control), CPF (5 mg/kg), MEDCL (100 mg/kg) and MEDCL (100 mg/kg) + CPF (5 mg/kg), following 7 days of acclimatization. After 4 weeks of treatments, the rats were sacrificed, and the levels of Superoxide dismutase (SOD), Catalase (CAT), Malondialdehyde (MDA), reduced glutathione (GSH), Glutathione peroxidase (GPx), glutathione S-transferase (GST) and DNA fragmentation were spectrophotometrically assessed in the brain and heart, while Acetylcholinesterase (AChE) activities were assessed in the serum and brain of the rats. Results: The results revealed that CPF significantly reduced the levels of SOD, CAT, GSH, GPx and GST, while that of MDA was elevated in brain and heart, compared with controls. The CPF significantly lowered AChE activities in serum and brain by 94% and 48% respectively, while the level of DNA fragmentation was significantly increased in the rats treated with CPF. Supplementation with MEDCL significantly ameliorated the changes in the rats. Conclusion: From the foregoing, the suppressive potential of methanol extract of Diospyros chloroxylon leaf is marked indicated in brain and cardiac redox imbalance induced on exposure to Chlorpyrifos. 


Fang, H., Y.L. Yu., W. Wang., M. Shan., X.M. Wu. and J.Q. Yu, 2006. Dissipation of chlopyrifos in pakchoi-vegetated soil in a greenhouse. J. Environ. Sci., 18: 760-764. PMID: 17078557.

Anderson, D.J, and R.A. Hites, 1988. Chlorinated pesticides in indoor air. Environ. Sci. Technol., 22 (6): 717-720.

Mackay, D., J.P. Giesy. and K.R. Solomon, 2014. Fate in the environment and long-range atmospheric transport of the organophosphorous pesticide, chlopyrifos and its oxon. Rev. Environ. Contam. Toxicol., 231: 35-76. doi: 10.1007/978-3-319-03865-0_3.

Sultatos, LG. and S.D. Murphy, 1983. Kinetic analyses of the microsomal biotransformation of the phosphotprothioate insecticides, chlopyrifos and parathion. Fundam. Appl. Toxicol., 3(1): 16.

Ma, T and J.E. Chambers, 1994. Kinetic parameters of desulfuration and dearylation of parathion and chlopyrifos by rat liver microsomes. Food Chem. Toxicol., 32(8): 763-767.

Giesy, J.P, K.R. Solomon., J.R. Coates., K.R. Dixon., J.M. Giddings. and E.E. Kenaga, 1999. Chlopyrifos: ecological risk assessment in North American aquatic environments. Rev. Environ. Contam. Toxicol., 160: 1-129. doi: 10.1007/978-3-319-03865-0_5

United State of America Environmental Protection Agency (USAEPA), 1999. Interim guidance on honey bee data requirements. United States Environmental Protection Agency, Environmental fate and effects division, Office of pesticide programs, Washington, DC.

Bakke, J.E, V.J Fell. and C.E. Price, 1976. Rat urinary metabolites FROM O, O-diethyl-O (3, 5, 6-trichloro-2-pyridyl) phosphorothioate. J. Environ Sci. Health Bull., 3: 225-230.

Nolan, RJ., D.L. Rick. and N.L. Freshour, 1984. Chlorpyrifos pharmacokinetics in human volunteers. Toxicol. Appl. Pharmacol., 73(1): 8-15. PMID: 6200956, DOI: 10.1016/0041-008x(84)90046-2

Chambers, J.E. and H.W. Chambers, 1989. Oxidative desulfuration of Chlorpyrifos, chlorpyrifos-methyl and leptophos by rat brain and liver. J. Biochem. Toxicol., 4 (3): 201-203.

Amer, S.M. and F.A.E. Aly, 1992. Cytogenic effects of pesticides IV. Cytogenic effects of the inse3cticides, Gardona and Dursban. Mutat. Res., 279 (3): 165-170. PMID: 1377331, DOI: 10.1016/0165-1218(92)90063-6.

Sobti, R.C., A. Krishan. and C.D. Pfaffenberger, 1982. Cytokinetic and cytogenetic effects of some agricultural chemicals on human lymphoid cells in vitro: Organophosphates. Mutat Res., 102: 89-102.

Ali, D., N.S. Nagpure., R. Kumar. and B. Kushwaha, 2008. Genotoxicity assessment of acute exposure of chlorpyrifos to freshwater fish Channa punctatus (Bloch) using micronucleus assay and alkaline single-cell electrophoresis. Chemosphere., 7: 1823-1831. DOI: 10.1016/j.chemosphere.2008.02.007.

Yin, X.H., G.N. Zhu., X.B. Li. and S.Y. Liu, 2009. Genotoxicity evaluation of chlorpyrifos to amphibian Chinese toad (Amphibian: Anura) by Comet assay and micronucleus test. Mutat. Res./Genetic Toxicol Environ Mutagen., 680 (1-2): 2-6. PMID: 19524702, DOI: 10.1016/j.mrgentox.2009.05.018

Cui, Y., J. Guo. and Z. Chen, 2011. Genotoxicity of chlorpyrifos and cypermethrin in ICR mouse lymphocytes. Toxicol Mech. Meth., 21(1): 70-74.

United State of America Environmental Protection Agency (USAEPA), 2009. Chlrpyrifos summary Document Registration Review: Initial Docet Number; EPA-HQ-OPP-2008-0850. United States Environmental Protection Agency, Washington, D.C.

Alavanja, M.C.R., C. Samanic., M. Dosemeci., J. Lubin., R. Tarone. and C.F.Lynch, 2003. Use of Agricultural pesticides and prostate cancer risk in the Agricul;tural Heaith Study cohort. Am. J. Epidemiol., 157(9): 800-814. PMID:12727674, DOI: 10.1093/aje/kwg040.

Engel, L.S., D.A. Hill., J.H. Lubin., C.F. Lynch., J. Pierce. and C. Samanic, 2005. Pesticide use and breast cancer risk among farmer’s wives in the Agricultural Health Study. Am. J. Epiemiol., 161(2): 121-135. PMID: 15632262, DOI: 10.1093/aje/kwi022.

Ventura, C., M. Nunez., N. Miret., D.M. Lamas., A. Randi. and A. Venturino, 2012. Differential mechanisms of actions are involved in chlorpyrifos effects in estrogen-dependent or – independent breast cancer cells exposed to low or high concentrations of the pesticide. Toxicol. Letts., 2213: 184-193. doi: 10.1016/j.toxlet.2012.06.017.

Lee, W.J., D.P. Sandler., A. Blair., C. Samanic., A.J. Cross and M.C.R. Alavanja, 2007. Pesticide use and colorectal cancer risk in the Agricultural Health Study. Int, J. Cancer. 121: 339-346. DOI: 10.1002/ijc.22635

Usmani, K.A., R.L. Rose. and E. Hodgson, 2003. Inhibition and activation of the human liver and human cytochrome p450 3A4 metabolism of testosterone by deployment-related chemicals. Drug Metabol. Disp., 31: 384-391. PMID: 12642463, DOI: 10.1124/dmd.31.4.384.

Usmani, K.A., T.M. Cho., R.L. Rose. and E. Hodgson, 2006. Inhibition of the human liver microsomal and human cytochrome p450 1A2 and 3A4 metabolism of estradiol by deployment-related and other chemicals. Drug Metabol Disp., 34: 1606-1614. PMID: 16790556, DOI: 10.1124/dmd.106.010439.

Viswanath, G., S. Chatterjee., S. Dabral., S.R. Nanguneri., G. Divya. and P. Roy, 2010. Anti-androgenic endocrine disrupting activities of chlorpyrifos and piperophos. J. Steroid Biochem. Mol. Biol., 120: 22-29. doi: 10.1016/j.jsbmb.2010.02.032.

Yonemori, K., A. Sugimura. and M. Amada, 2000. Persimmon genetics and breeding. Plant Breed. Rev., 19: 191-225.

Adzu, B., S. Amos., S. Dzarma., I. Muazzam. and K.S. Gamaniel, 2002. Pharmacological evidence favouring the folkloric use of Diospyros mespiliformis Hochst in the relief of pain and fever. J Ethnopharmacol., 82:191-195. PMID: 12241995, DOI: 10.1016/s0378-8741(02)00179-4

Trongsakul, S., A. Panthong., D. Kanjanapothi. and T. Taesotikul, 2003. The analgesic, antipyretic and anti-inflammatory activities of Diospyros variegate Kruz. J. Ethnopharmacol., 85: 221-225. PMID: 12639744, DOI: 10.1016/s0378-8741(03)00020-5

Reddy, K.N, G. Trimurthulu., and S.C. Reddy, 2008. Medicinal plants used by ethnic people of Medak district, Andhra Pradesh. Indian J. Traditional Knowledge. 9(1): 184-190.

Kim, D.S., Z. Chen., V.T. Nguyen., J.M. Pezzuto. and L.U. Qius, 1997. A concise semi-synthetic approach to Betulinic acid from Betulin. Synth commun., 27:1607.

Higa, M., K. Ogihara. and N. Yogis, 1998. Bioactive naphthoquinone derivatives from dispyros maritime blume. Chem. Pharm bull., 46:1189-1193

Ehrhardt, H., S. Fulda., M. Fuhrer., K.M. Debatin. and I. Jeremias, 2004. Betulinic acid induced apoptosis in leukemia cells. Leukemia. 18:1406-1412. PMID: 15201849, DOI: 10.1038/sj.leu.2403406.

Fulda, S, 2008. Betulinic acid for cancer treatment and prevention. Int. J. Mol. Sci., 9:1096-1107. PMID: 19325847, DOI: 10.3390/ijms9061096

Chintharlapalli, S, S. Papineni., P. Lei., S. Pathi. and S. Safe, 2011. Betulinic acid Inhibits colon cancer cell and tumor growth and induces proteasome: dependent and independent down regulation of specificity proteins (sp) transcription factors. BMC Cancer. 11, 371-382. PMID: 21864401, DOI: 10.1186/1471-2407-11-371

Adeleke, G.E. and O.A. Adaramoye, 2016. Modulatory role of Betulinic acid in N-nitrosodimethy lamine-induced toxicity in male rats. Hum and Exper Toxicol., 1-10. doi: 10.1177/0960327116661399.

Adeleke, G.E. and O.A. Adaramoye, 2017. Betulinic acid protects against N-nitrosodimethylamine-induced redox imbalance in testes of rats. Redox Rep., 22(6): 556-562. doi: 10.1080/13510002.2017.1322750.

Adeleke, G.E., J.A. Badmus., O.T. Adedosu., D.H. Abioye and O.O. Arinde, 2018. Betulinic acid attenuates hepatic and testicular redox imbalance and DNA damage in male rats exposed to crude oil. J. Pharmacog and Phytochem., 7(3): 316-322.

Adeleke, G.E., O.T. Adedosu., O.A. Adaramoye., A.S. Olagunju., O.J. Teibo., O.O. Arinde. and O.K. Afolabi, 2016. Hepatoprotective effect of methanol extract of Diospyros chloroxylon leaf in N-Nitrosodimethylamine-induced hepatotoxicity in Wistar rats. As J. Health Sci., 3 (3): 142-152. e-ISSN: 2349-0659, p-ISSN: 2350-0964

Adeleke, G.E, O.T. Adedosu., O.K. Afolabi,, O.O. Arinde. and T.M. Oyedokun, 2017. Methanolic extract of Diospyros chloroxylon modulates hepatic redox profile and cell proliferation in Dimethylamine-treated rats. Br. J. Med and Med. Res., 21(2): 1-12. DOI: 10.9734/BJMMR/2017/33242.

Lowry, O.H, 1951. Rosbrough NJ, Farr AL. Protein measurement with the Folin-phenol reagent. J. Biol. Chem., 193, 265-275. PMID: 14907713.

Ohkawa, H., N. Ohishi. and K. Yagi, 1979. Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem., 95: 351-358. ISSN : 0003-2697.

Misra, H.P. and J. Fridovich, 1972. The role of superoxide anion in the autoxidation of epinephrine and a simple assay for superoxide dismutase. J. Biol. Chem., 247: 3170-3175. PMID: 4623845.

Aebi, H, 1984. Catalase in vitro. In: Packer L. Editor. Methods in Enzymology. Orlando FL: Academic Press. p. 121-126.

Andersen, H.R., J.B. Nielsen. and F. Nielsen, 1997. Antioxidative enzyme activities in human erythrocytes. Clin. Chem., 43 (4): 562-568. PMID:9105255.

Mitchell, J.R., D.J. Jollow. and W.Z. Potter, 1973. Acetaminophen-induced hepatic necrosis I Role of drug metabolism. J Pharmacol Expt Therap., 87:185-194. PMID:4746327.

Habig, W., M. Pabst. and W. Jakoby, 1974. Glutathione S-transferases: The first enzymatic step in mercapturic acid formation. J Biol Chem., 249: 7130–7139. PMID: 4436300.

Ellman, G.L., K.D. Courtney., V. Jr. Andres. and Y. Featherstone, 1961. A new and rapid colorimetric determination of acetylcholinesterase activity. Biohem. Pharmacol., 7: 88-95. PMID:13726518. DOI: 10.1016/0006-2952(61)90145-9.

Wu, B., A. Ootani., R. Iwakiri., Y. Sakata., T. Fujise., S. Amemori., F. Yokoyama., S. Tsunada . and K. Fujimoto, 2005. T-cell deficiency leads to liver carcinogenesis in Azoxymethane-treated rats. Exp Biol Med., 231: 91-98.

Peerera, F.P., V. Rauh., W.Y. Tsai., P. Kinney., D. Camann., D. Barr., T. Bernert., R. Garfinkel. and Y.H. Tu, 2003. Effects of transplacental exposure to environmental pollutants on birth outcomes in a multiethnic population. Environ. Health Perspect., 111(2): 201-205. doi: 10.1289/ehp.5742.

Whyatt, R.M, V. Rauh., D.B. Barr., D. Camann., H.F. Andrews., R. Garginkel., L.A. Hoepner., D. Diaz. and J. Dietrich, 2004. Prenatal insecticide exposures and birth weight and length among an urban minority cohort. Environ. Health Perspect., 112(10): 1125-1132. DOI: 10.1289/ehp.6641.

Neuget, A.L., M. Hayek. and G. Howe, 1996. Epidemiology of gastic cancer. Semin oncol., 23: 281. PMID: 8658212

Allen, R.G. and M. Tresini, 2000. Oxidative stress and gene regulation. Free Radic. Biol. Med., 28, 463-499. DOI: 10.1016/s0891-5849(99)00242-7.

Toyokuni, S., 2006. Novel aspects of oxidative stress-associated carcinogenesis. Antioxid. Redox. Signal. 8, 1373-1377.

Ishikawa, K., K. Takenaga., M. Akimoto., N. Koshikawa., A. Yamaguchi.,, H. Imanishi., K. Nakada, Y. Honma. and J. Hayashi, 2008. ROS-generating mitochondrial DNA mutations can regulate tumor cell metastasis. Sci., 320: 661-664. doi: 10.1126/science.1156906.

Kumar, B., S. Koul., L. Khandrika., R.B. Meacham. and H.K. Koul, 2008. Oxidative stress is inherent in prostate cancer cells and is required for aggressive phynotype. Cancer Res., 68: 1777-1785. doi: 10.1158/0008-5472.

Demir, E., B. Kaya. and C. Soriano, 2011. Genotoxic analysis of four lipid-peroxidation products in the mouse lymphoma assay. Mutat. Res., 726: 98–103. doi: 10.1016/j.mrgentox.2011.07.001.

Mehta, A., .R.S. Verma. and N. Srivastava, 2009. Chlorpyrifos induced alterations in the levels of hydrogen peroxide, nitrate and nitrite in rat brain and liver. Pestic Biochem. Physiol., 94(2-3): 55-59.

Verma, R.S., A. Mehta. and N. Srivastava, 2009. Comparative studies on Chlorpyrifos- and methylparathion- induced oxidative stress in different parts of rat brain: Attenuation by antioxidant vitamins. Pestic Biochemi Physiol., 95(3): 152-158. DOI: 10.1016/j.pestbp.2009.08.004

Mansour, S.A. and A. Mossa, 2009. Lipid peroxidation and oxidative stress in rat erythrocytes induced by chlorpyrifos and protective effect of Zinc. Pestic Biochem Physiol.,93(1): 34-39.

Kalendar, Y., S. Kaya., D. Durak., F.G. Uzun. and F. Demir F, 2012. Protective effects of catechin and quercetin on antioxidant status, lipid peroxidation and testis-histoarchitecture induced by chlorpyrifos in male rats. Environ Toxicol Pharmacol., 32(2): 141-148. doi: 10.1016/j.etap.2011.12.008.

Attia, A.A., R.H. ElMazoudy. and N.S. El-Shenawy, 2012. Antioxidant role of propolis extract against oxidative damage of testicular tissue induced by insecticide chlorpyrifios in rats. Pestic Biochem Physiol., 103: 87-93.

Elsharkawy, E.E., D. Yahia. and N.A. ElNisr, 2013. Sub-chronic exposure to chlorpyrifos induces heamatological, metabolic disorders and oxidative stress in rat: attenuation by glutathione. Environ Toxicol Pharmacol., 35(2): 218-227. doi: 10.1016/j.etap.2012.12.009.

Bebe, F.N. and M. Panemangalore, 2013. Exposure to low doses of endosulfan and chlorpyrifos modifies endogenous antioxidants in tissues of rats. J. Environ Sci. Health B 38(3): 349-363. DOI: 10.1081/PFC-120019901

Hunt, C.R., J.E. Sim., S.J. Sullivan., T. Featherstone, C. Von Kapp-Herr., R.A. Hock., R.A. Gomez., A.J. Parsian. and D.R. Spitz, 1998. Genomic instability and catalase gene amplification induced by chronic response to oxidative stress. Cancer Res., 58: 3986-3992. PubMed 9731512

Arthur, J.R., 2000. The glutathione peroxidases. Cell Moll Life Sci., 57: 1825-1835. DOI: 10.1007/pl00000664

Lauterburg, B.H. and M.E. Velez, 1998. Glutathione deficiency in alcoholics: risk factor for paracetamol hepatotoxicity. Gut., 29(9): 1153-1157. doi: 10.1136/gut.29.9.1153.

Edwards, R., D.P. Dixon. and V. Walbot, 2000. Plant glutathione S-transferases: enzymes with multiple functions in sickness and in health. Trends in plant Sci., 5 (5): 193-198. DOI: 10.1016/s1360-1385(00)01601-0

Binková, B., P. Strejc., O. Boubelík., Z. Stávková., I. Chvátalová. and R.J. Srám, 2001. DNA adducts and human atherosclerotic lesions. International Journal of Hygene and Environmental Health., 204 (1): 49–54.

Hodgson, E. and R.L. Rose, 2008. Metabolic interactions of Agrochemicals in humans. Pest Manag Sci., 64(6): 617-621. doi: 10.1002/ps.1563.

Flakos, J, 2012. The developmental neurotoxicity of organophosphorus insecticides: A direct role for the oxon metabolites. Toxicol Lett., 209(1): 86-93. doi: 10.1016/j.toxlet.2011.11.026.

Li, J., G. Pang., F. Ren. and B. Fang, 2019. Chlorpyrifos-induced reproductive toxicity in rats could bee partly relieved under high-fat diet. Chemosphere. 229: 94-102. Doi: 10.1016/j. Chemosphere. 2019.05.020.

Kaplan, J.G., J. Kessler. and N. Rosenberg, 1993. Sensory neuropathy associated with Dursban (Chlorpyrifos) exposure. Neurol., 43(11): 2193-2196.

Gotoh, M., I. Saito., J. Huang., Y. Fukaya., T. Matsumoto., N. Hosanaga., E. Shibata., G. Ichihara., M. Kamujima. and Y. Takeuchi, 2001. Chnges in cholinesterase activity, nerve conduction velocity, and clinical signs and symptoms in termite control operators exposed to chlorpyrifos. J. Occup. Health., 43: 157-164. DOI: 10.1539/joh.43.157.

Farag, A.T., A.M El Okazy. and A.F. El-Aswd, 2003. Developmental toxicity study of chlorpyrifos in rats. Repo Toxicol., 17: 203-208. DOI: 10.1016/s0890-6238(02)00121-1.

Cutler, G.C., J. Purdy., J.P. Giesy. and K.R. Solomon, 2014. Risk to pollinators from the use of chlorpyrifos in the United States. Rev. Environ Contam Toxicol., 231: 219-265. doi: 10.1007/978-3-319-03865-0_7.

Colović, M.B., D.C. Krstić., T.D. Lazarević-Pašti., A.M. Bondžić.. and V.M. Vasić, 2013. Acetylcholinesterase inhibitors: pharmacology and toxicology. Current neuropharmacol., 11(3):315-35. doi:10.2174/1570159X11311030006

Song, X., J.D. Violin., F.J. Seidler. and T. Slotkin, 1998. Modeling the developmental neurotoxicity of chlorpyrifos in vitro: macromolecule synthesis in PC12 cells. Toxicol Appl Pharmacol., 151(1): 182-191. DOI: 10.1006/taap.1998.8424

Slotkin, T.A, 2004. Guidlines for developmental neurotoxicity and their impact on organophosphate pesticides: a personal view from an academic perspective. Neurotoxicol., 25(4): 631-640. DOI: 10.1016/S0161-813X(03)00050-0.

Slotkin, T.A., E.D. Levin. and F.J. Seidler, 2006. Comparative developmental neurotoxicity of organophosphate insecticides: effects on brain development are separable from systemic toxicity. Environ Health Perspect, 114: 746-751. doi: 10.1289/ehp.8828.

Qiao, D., F.J. Seidler., S. Padilla. and T.A. Slotkin, 2002. Developmental neurotoxicity of chlorpyrifos: What is the vulnerable period? Environ Health Perspect., 110: 1097-1103. doi: 10.1289/ehp.021101097.

California Environmental Protection Agency, 2008. Evidence on the developmental and reproductive toxicity of chorpyrifos.Reproductive and Cancer hazard Assessment Branch, Office of environmental Health Hazard Assessment, California Environmental Protection Agency.

Sandoval, L., A. Rosca., A. Oniga., A. Zambrano., J.J. Ramos., M.C. Gonzelez., I. Liste. and M. Motas, 2019. Effects of chlorpyrifos on cell death and cellular phenotypic specification of human neural stem cells. Sci. Total Environ., 683: 445-454. Doi. 10.1016/j. scitotenv.2019.05.270.

Lieberman, A.D., M.R. Craven., H.A. Lewis., J.H. Nemenzo, 1998. Genotoxicity from domestic use of organophosphate pesticides. J. Occup Environ Med., 40(11): 954-957. DOI: 10.1097/00043764-199811000-00003

Yu, F., Z. Wang., B. Ju., J. Wang. and D. Bai, 2008. Apoptotic effects of organophosphorus insecticide chlorpyrifos on mouse retina in vivo via oxidative stress and protection of combination of vitamins C and E. Exp Toxicol Pathol., 59(6): 415-423. doi: 10.1016/j.etp.2007.11.007] [pmid: 18222074].




How to Cite

Adeleke, G. E., Adedosu, O. T., Arinde, O. O., Adeyemo, O. E., Bolarinwa, W., & Oyewo, E. B. (2021). Methanol Leaf Extract of <i>Diospyros chloroxylon</i> Attenuates Chlorpyrifos-Induced Toxicity in Rats. Innoriginal: International Journal of Sciences, 8(2), 34–46. Retrieved from