Efficacy of crude extract of Emblica officinalis (amla) in arsenic‑induced oxidative damage and apoptosis in splenocytes of mice


Affiliations

  • King George Medical University, Department of Pharmacology
  • Dr. Harisingh Gour Central University, Department of Criminology and Forensic Science, School of Applied Sciences, Dr. Harisingh Gour Central University, Sagar, Madhya Pradesh, India
  • CSIR‑Indian Institute of Toxicology Research, Proteomics Laboratory, Lucknow, Uttar Pradesh, India

Abstract

Introduction: Arsenic, an environmental contaminant naturally occurred in groundwater and has been found to be associated with immune‑related health problems in humans. Objective: In view of increasing risk of arsenic exposure due to occupational and non‑occupational settings, the present study has been focused to investigate the protective efficacy of amla against arsenic‑induced spleenomegaly in mice. Results: Arsenic exposures (3 mg/kg body weight p.o for 30 days) in mice caused an increase production of ROS (76%), lipid peroxidation (84%) and decrease in the levels of superoxide dismutase (53%) and catalase (54%) in spleen as compared to controls. Arsenic exposure to mice also caused a significant increase in caspases‑3 activity (2.8 fold) and decreases cell viability (44%), mitochondrial membrane potential (47%) linked with apoptosis assessed by the cell cycle analysis (subG1‑28.72%) and annexin V/PI binding in spleen as compared to controls. Simultaneous treatment of arsenic and amla (500 mg/kg body weight p.o for 30 days) in mice decreased the levels of lipid peroxidation (33%), ROS production (24%), activity of caspase‑3 (1.4 fold), apoptosis (subG112.72%) and increased cell viability (63%), levels superoxide dismutase (80%), catalase (77%) and mitochondrial membrane potential (66%) as compared to mice treated with arsenic alone. Conclusions: Results of the present study indicate that the effect of arsenic is mainly due to the depletion of glutathione in liver associated with enhanced oxidative stress that has been found to be protected following simultaneous treatment of arsenic and amla.

Keywords

Amla, apoptosis, arsenic, mice, oxidative stress, spleen

Full Text:

References

Leonard SS, Harris GK, Shi X. Metal‑induced oxidative stress and signal transduction. Free Radic Biol Med 2004;37:1921‑42.

WHO (World Health Organization). Arsenic and Arsenic Compounds. Inter‑organization Programm for the Sound Management of Chemicals. 2nd ed.. Environmental Health Criteria 224. Geneva: World Health Organization; 2001.

Yadav RS, Sankhwar ML, Shukla RK, Chandra R, Pant AB, Islam F, et al. Attenuation of arsenic neurotoxicity by curcumin in rats. Toxicol Appl Pharmacol 2009;240:367‑76.

Andrew AS, Jewell DA, Mason RA, Whitfield ML, Moore JH, Karagas MR. Drinking‑water arsenic exposure modulates gene expression in human lymphocytes from a U.S. population. Environ Health Perspect 2008;116:524‑31.

Vega L, Montes de Oca P, Saavedra R, Ostrosky‑Wegman P. Helper T cell subpopulations from women are more susceptible to the toxic effect of sodium arsenite in vitro. Toxicology 2004;199:121‑8.

Martin‑Chouly C, Morzadec C, Bonvalet M, Galibert MD, Fardel O, Vernhet L. Inorganic arsenic alters expression of immune and stress response genes in activated primary human T lymphocytes. Mol Immunol 2011;48:956‑65.

Stepnik M, Stańczyk M, Arkusz J, Lewińska D. Assessment of apoptosis in thymocytes and splenocytes from mice exposed to arsenate in drinking water: Cytotoxic effects of arsenate on the cells in vitro. J Environ Sci Health A Tox Hazard Subst Environ Eng 2005;40:369‑84.

Moore SE, Prentice AM, Wagatsuma Y, Fulford AJ, Collinson AC, Raqib R, et al. Early‑life nutritional and environmental determinants of thymic size in infants born in rural Bangladesh. Acta Paediatr 2009;98:1168‑75.

Rahman A, Vahter M, Ekström EC, Persson LÅ. Arsenic exposure in pregnancy increases the risk of lower respiratory tract infection and diarrhea during infancy in Bangladesh. Environ Health Perspect 2011;119:719‑24.

Mc‑Collum G, Keng PC, States JC, McCabe MJ Jr. Arsenite delays progression through each cell cycle phase and induces apoptosis following G2/M arrest in U937 myeloid leukemia cells.J Pharmacol Exp Ther 2005;313:877‑87.

Tenorio EP, Saavedra R. Differential effect of sodium arsenite during the activation of human CD4+and CD8+T lymphocytes.Int Immunopharmacol 2005;5:1853‑69.

Soto‑Peña GA, Luna AL, Acosta‑Saavedra L, Conde P, López‑Carrillo L, Cebrián ME, et al. Assessment of lymphocyte subpopulations and cytokine secretion in children exposed to arsenic. FASEB J 2006;20:779‑81.

Biswas R, Ghosh P, Banerjee N, Das JK, Sau T, Banerjee A, et al. Analysis of T‑cell proliferation and cytokine secretion in the individuals exposed to arsenic. Human Exp Toxicol 2008;27:381‑6.

Hernández‑Castro B, Doníz‑Padilla LM, Salgado‑Bustamante M, Rocha D, Ortiz‑Pérez MD, Jiménez‑Capdeville ME, et al. Effect of arsenic on regulatory T cells. J Clin Immunol 2009;29:461‑9.

Fry RC, Navasumrit P, Valiathan C, Svensson JP, Hogan BJ, Luo M, et al. Activation of inflammation/NF‑kappaB signaling in infants born to arsenic‑exposed mothers. PLoS Genet 2007;3:e207.

Aggarwal M, Naraharisetti SB, Dandapat S, Degen GH, Malik JK.Perturbations in immune responses induced by concurrent subchronic exposure to arsenic and endosulfan. Toxicology 2008;251:51‑60.

Pillai CK, Pillai KS. Antioxidants in health. Indian J Physiol Pharmacol 2002;46:1‑5.

Vahter M. Effects of arsenic on maternal and fetal health. Annu Rev Nutr 2009;29:381‑99.

Chowdhury R, Chatterjee R, Giri AK, Mandal C, Chaudhuri K.Arsenic‑induced cell proliferation is associated with enhanced ROS generation, ERK signaling and CyclinA expression. Toxicol Lett 2010;198:263‑71.

Flora SJ. Arsenic‑induced oxidative stress and its reversibility.Free Radic Biol Med 2011;51:257‑81.

Ling YH, Jiang JD, Holland JF, Perez‑Soler R. Arsenic trioxide produces polymerization of microtubules and mitotic arrest before apoptosis in human tumor cell lines. Mol Pharmacol 2002;62:529‑38.

Patra PH, Bandyopadhyay S, Bandyopadhyay MC, Mandal TK.Immunotoxic and genotoxic potential of arsenic and its chemicals species in goats. Toxicol Int 2013;20:6‑10.

Singh MK, Yadav SS, Gupta V, Khattri S. Immunomodulatory role of Emblica officinalis in arsenic induced oxidative damage and apoptosis in thymocytes of mice. BMC Complement Altern Med 2013;13:193.

Singh MK, Dwivedi S, Yadav SS, Sharma P, Khattri S Arsenic‑induced hepatic toxicity and its attenuation by fruit extract of Emblica‑officinalis (Amla) in mice. Indian J Clin Biochem 2014;29;29-37

Oluwaseun AA, Ganiyu O. Antioxidant properties of methanolic extracts of mistletoes (Viscum album) from cocoa and cashew trees in Nigeria. Afr J Biotechnol 2008;7:3138‑42.

Ihantola‑Vormisto A, Summanen J, Kankaanranta H, Vuorela H, Asmawi ZM, Moilanen E. Anti‑inflammatory activity of extracts from leaves of Phyllanthus emblica. Planta Med 1997;63:518‑24.

Riccioni G, Bucciarelli T, Mancini B, Di Ilio C, Capra V, D’Orazio N. The Role of the antioxidant vitamin supplementation in the prevention of cardiovascular diseases. Expert Opin Investig Drugs 2007;16:25‑32.

Poltanov EA, Shikov AN, Dorman HJ, Pozharitskaya ON, Makarov VG, Tikhonov VP, et al. Chemical and antioxidant evaluation of Indian gooseberry (Emblica officinalis Gaertn., syn. Phyllanthus emblica L.) supplements. Phytother Res 2009;23:1309‑15.

Ogasawara M, Matsunaga T, Suzuki H. Differential effects of antioxidants on the in vitro invasion, growth and lung metastasis of murine colon cancer cells. Biol Pharm Bull 2007;30:200‑4.

Suresh K, Vasudevan DM. Augmentation of murine natural killer cell and antibody dependent cellular cytotoxicity activities by Phyllanthus emblica, a new immunomodulator. J Ethnopharmacol 1994;44:55‑60.

Liu X, Cui C, Zhao M, Wang J, Luo W, Yang B, et al. Identification of phenolics in the fruit of emblica (Phyllanthus emblica L.) and their anti‑oxidant activities. Food Chem 2008;109:909‑15.

Luo W, Zhao M, Yang B, Shen G, Rao G. Identification of bioactive compounds in Phllenthus emblica L. fruit and their free radical scavenging activities. Food Chem 2009;114:499‑504.

Schulz H, Nagymajtényi L, Institoris L, Papp A, Siroki O. A study on behavioral, neurotoxicological, and immunotoxicological effects of subchronic arsenic treatment in rats. J Toxicol Environ Health A 2002;65:1181‑93.

Khandelwal S, Shukla LJ, Shanker R. Modulation of acute Cadmium toxicity by Emblica officinalis fruit in rat. Indian J Exp Biol 2002;40:564‑70.

Suja RS, Nair AM, Sujith S, Preethy J, Deepa AK. Evaluation of immunomodulatory potential of Emblica officinalis fruit pulp extract in mice. Indian J Anim Res 2009;43:103‑6.

Sharma A, Sharma MK, Kumar M. Modulatory role of Emblica officinalis fruit extract against arsenic induced oxidative stress in Swiss albino mice. Chem Biol Interact 2009;180:20‑30.

Gao S, Wang Y, Zhang P, Dong Y, Li B. Subacute oral exposure to dibromoacetic acid induced immunotoxicity and apoptosis in the spleen and thymus of mice. Toxicol Sci 2008;105:331‑41.

Mosmann T. Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays.J Immunol Methods 1988;65:55‑63.

Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem 1951;193:263‑75.

Wang JF, Jerrells TR, Spitzer JJ. Decreased production of reactive oxygen intermediates is an early event during in vitro apoptosis of rat thymocytes. Free Radic Biol Med 1996;20:533‑42.

Ohkawa H, Osishi N, Yagi K. Assay for lipid peroxides in animal tissue by thiobarbituric acid reaction. Anal Biochem 1979;95:351‑8.

Marklund S, Marklund G. Involvement of superoxide anion radical in the autooxidation of pyrogallol and a convenient assay for superoxide dismutase. Eur J Biochem 1974;47:469‑74.

Bai J, Rodriguez AM, Melendez JA, Cederbaum AI. Overexpression of catalase in cytosolic or mitochondrial compartment protects HepG2 cells against oxidative injury. J Biol Chem 1999;274:26217‑24.

Darzynkiewicz Z, Juan G, Li X, Gorczyca W, Murakami T, Traganos F. Cytometry in cell necrobiology: Analysis of apoptosis and accidental cell death (necrosis). Cytometry 1997;27:1‑20.

Vermes I, Haanen C, Steffens‑Nakken H, Reutelingsperger C.A novel assay for apoptosis. Flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein labelled Annexin V. J Immunol Methods 1995;184:39‑51.

Burchiel SW, Mitchell LA, Lauer FT, Sun X, McDonald JD, Hudson LG, et al. Immunotoxicity and biodistribution analysis of arsenic trioxide in C57Bl/6 mice following a 2‑wk inhalation exposure. Toxicol Appl Pharmacol 2009;241:253‑9.

Li Q, Lauer FT, Liu KJ, Hudson LG, Burchiel SW. Low‑dose synergistic immunosuppression of T‑dependent antibody responses by polycyclic aromatic hydrocarbons and arsenic in C57BL/6J murine spleen cells. Toxicol Appl Pharmacol 2010;245:344‑51.

Das S, Pan D, Bera AK, Rana T, Bhattacharya D, Bandyapadyay S, et al. Sodium arsenite mediated immune‑disruption through alteration of transcription profile of cytokines in chicken splenocytes under in vitro system. Mol Biol Rep 2011;38:171‑6.

Mishra D, Mehta A, Flora SJ. Reversal of arsenic‑induced hepatic aoptosis with combined administration of DMSA and its analogues in guinea pigs: Role of glutathione and linked enzymes. Chem Res Toxicol 2008;21:400‑7.

Habib A, Hayashi T, Sato KK, Hata A, Ikebe M, Rahman F, et al. Effectiveness of arsenic mitigation program in Bangladesh‑‑relationship between arsenic concentrations in well water and urine. Osaka City Med J 2007;53:97‑103.

Pulido MD, Parrish AR. Metal‑induced apoptosis: Mechanisms.Mutat Res 2003;533:227‑41.

Fulda S, Debatin KM. Extrinsic versus intrinsic apoptosis pathways in anticancer chemotherapy. Oncogene 2006;25:4798‑811.

Cadenas E. Mitochondrial free radical production and cell signaling. Mol Aspects Med 2004;25:17‑26.

Hossain K, Akhand AA, Kato M, Du J, Takeda K, Wu J, et al. Arsenite induces apoptosis of murine T lymphocytes through membrane raft‑linked signaling for activation of c‑Jun amino‑terminal kinase. J Immunol 2000;165:4290‑7.

Li D, Morimoto K, Takeshita T, Lu Y. Formamidopyrimidine‑DNA glycosylase enhances arsenic‑induced DNA strand‑breaks in PHA‑stimulated and unstimulated human lymphocytes. Environ Health Perspect 2001;109:523‑6.

Maeda H, Hori S, Nishitoh H, Ichijo H, Ogawa O, Kakehi Y, et al.Tumor growth inhibition by arsenic trioxide (As2O3) in the orthotopic metastasis model of androgen‑independent prostate cancer. Cancer Res 2001;61:5432‑40.

Banerjee N, Banerjee M, Ganguly S, Bandyopadhyay S, Das JK, Bandyopadhay A, et al. Arsenic‑induced mitochondrial instability leading to programmed cell death in the exposed individuals. Toxicology 2008;246:101‑11.

Flora S, Bhatt K, Mehta A. Arsenic moiety in gallium arsenide is responsible for neuronal apoptosis and behavioral alterations in rats. Toxicol Appl Pharmacol 2009;240:236‑44.

Sai‑Ram M, Neetu D, Yogesh B, Anju B, Dipti P, Pauline T, et al. Cyto‑protective and immunomodulating properties of Amla (Emblica officinalis) on lymphocytes: An in‑vitro study.J Ethnopharmacol 2002;81:5‑10.

Pramyothin P, Samosorn P, Poungshompoo S, Chaichantipyuth C.The protective effects of Phyllanthus emblica Linn. extract on ethanol induced rat hepatic injury. J Ethnopharmacol 2006;107:361‑4.

Anilkumar KR, Nagaraj NS, Santhanam K. Reduction of hexachlorocyclohexane‑induced oxidative stress and cytotoxicity in rat liver by Emblica officinalis Gaertn. Indian J Exp Biol 2007;45:450‑4.

Haque R, Bin‑Hafeez B, Ahmad I, Parvez S, Pandey S, Raisuddin S. Protective effects of Emblica officinalis Gaertn in cyclophosphamide‑treated mice. Human Exp Toxicol 2001;20:643‑50.

Bandyopadhyay SK, Pakrashi SC, Pakrashi A. The role of antioxidant activity of Phyllanthus emblica fruits on prevention from indomethacin induced gastric ulcer. J Ethnopharmacol 2000;70:171‑6.


Refbacks

  • There are currently no refbacks.