Gallic Acid Isolated from Pomegranate Peel Extract Induces Reactive Oxygen Species Mediated Apoptosis in A549 Cell Line
Santhini Elango, Ramji Balwas, Viswanadha Vijaya Padma
DOI: 10.4236/jct.2011.25085   PDF   HTML     6,336 Downloads   13,049 Views   Citations


Antioxidant properties elicited by plant species have a full range of perspective applications in human health care. In recent years, the prevention of cancer and cardiovascular diseases has been associated with the ingestion of fresh fruits, vegetables or teas rich in natural antioxidants [1]. Extracts from the different part of the pomegranate plant such as juice, seed and peel have been reported to exhibit a potent antioxidant activity. But the anticarcinogenic activity of active principle from the pomegranate peel extract was not studied so far. Hence the present study was planned to explore the molecular mechanism of the anticarcinogenic activity pomegranate peel on A549 cell line. In this study, GC-MS analysis was carried out for the methanolic extract of pomegranate peel which revealed gallic acid (GA) as the major antioxidant compound in the extract. Hence GA was purified further through RP-HPLC and evaluated its anticancer potential by studying its effect on mitochondrial respiration, cell-membrane integrity, apoptotic body formation and the DNA fragmentation in cultured A549 cells. We observed increased level of reactive oxygen species in the cells treated with GA at the concentrations of 10 and 20 ug/ml. Further analysis of caspase activation (caspase 8 and 9) revealed activation of caspases 9 in the cells treated with GA at a concentration of 20 ug/ml. Thus the present study revealed that the GA isolated from the pomegranate peel extract (Kabul variety) induced apoptosis in A549 cells through intrinsic pathway.

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S. Elango, R. Balwas and V. Padma, "Gallic Acid Isolated from Pomegranate Peel Extract Induces Reactive Oxygen Species Mediated Apoptosis in A549 Cell Line," Journal of Cancer Therapy, Vol. 2 No. 5, 2011, pp. 638-645. doi: 10.4236/jct.2011.25085.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] D. F. Wang, J. Li, C. H. Wang, G. W. Zhao, Y. Jin, D. D. Chen and S. Ye, “Study on the Component and Immune Activity of Polysaccharides from Tea,” Journal of Tea Science, Vol. 20, No. 1, 2000, pp. 45-50.
[2] T. Sugimura, “Food and Cancer,” Toxicology, Vols. 181-182, 2002, pp. 17-21.
[3] M. I. Gil, F. A. Tomas-Barberan, B. Hess-Pierce, D. M. Holcroft and A. A. Kader, “Antioxidant Activity of Pomegranate Juice and Its Relationship with Phenolic Composition and Processing,” Journal of Agricultural and Food Chemistry, Vol. 48, No. 10, 2000, pp. 4581-4589. doi:10.1021/jf000404a
[4] M. Aviram, L. Dornfield, M. Rosenblatt, N. Volkova, M. Kaplan, R. Coleman, et al., “Pomegranate Juice Consumption Reduces Oxidative Stress, Atherogenic Modifications to LDL, and Platelet Aggregation: Studies in Humans and in Atherosclerotic Apolipoprotein E-Deficient Mice,” American Journal of Clinical Nutrition, Vol. 71, No. 5, 2000, pp. 1062-1076.
[5] M. Kaplan, T. Hayek, A. Raz, R. Coleman, L. Dornfield, J. Vayan, et al., “Pomegranate Juice Supplementation to atherosclerotic Mice Reduces Macrophage Lipid Peroxidation, Cellular Cholesterol Accumulation and Development of Atherosclerosis,” Journal of Nutrition, Vol. 131, No. 8, 2001, pp. 2082-2089.
[6] N. D. Kim, R. Mehta, W. Yu, I. Neeman, T. Livney, A. Amichay, et al., “Chemopreventive and Adjuvant Therapeutic Potential of Pomegranate (Punica granatum) for Human Breast Cancer,” Breast Cancer Research and Treatment, Vol. 71, No. 3, 2002, pp. 203-217. doi:10.1023/A:1014405730585
[7] B. Cerda, J. J. Ceron, F. A. Tomas-Barberan and J. C. Espin, “Repeated Oral Administration of High Doses of Pomegranate Ellagitannin Punicalagin to Rats for 37 Days Is Not Toxic” Journal of Agricultural and Food Chemistry, Vol. 51, No. 11, 2003, pp. 3493-3501. doi:10.1021/jf020842c
[8] B. Cerda, R. Llorach, J. J. Ceron, J. C. Espin and F. A. Tomas-Barberan “Evaluation of the Bioavailability and Metabolism in the Rat of Punicalagin, an Antioxidant Polyphenol from Pomegranate Juice,” European Journal of Nutrition, Vol. 42, No. 1, 2003, pp. 18-28. doi:10.1007/s00394-003-0396-4
[9] A. K. Batta and S. Rangaswami, “Crystalline Chemical Components of Some Vegetable Drugs” Phytochemistry, Vol. 12, No. 1, 1973, pp. 214-216. doi:10.1016/S0031-9422(00)84654-3
[10] R. P. Singh, K. N. C. Murthy and G. K. Jayaprakasha, “Studies on Antioxidant Activity of Pomegranate (Punica granatum) Peel and Seed Extracts Using in Vitro Models,” Journal of Agricultural and Food Chemistry, Vol. 50, No. 1, 2002, pp. 81-86. doi:10.1021/jf010865b
[11] T. Mossman, “Rapid Colorimetric Assay for Cellular Growth of Survival, Application to Proliferation of Cytotoxicity Assays,” Journal of Immunological Methods, Vol. 65, No. 1-2, 1983, pp. 55-63. doi:10.1016/0022-1759(83)90303-4
[12] A. A. Nieland, “Lactic Dehydrogenase of Heart Muscle,” Academic Press, Cambridge, 1959.
[13] J. A. Royall and H. Ischiropoulos, “Evaluation of 2’, 7’-Dichlorofluorescein and Dihydrorhodamine 123 as fluorescent Probes for Intracellular H2O2 in Cultured Endothelial Cells,” Archives in Biochemistry and Biophysics, Vol. 302, No. 2, 1993, pp. 348-352. doi:10.1006/abbi.1993.1222
[14] M. S. Moron, J. W. Depierre and B. Mannervik, “Levels of Glutathione, Glutathione Reductase and Glutathiones S-Transferase Activities in Rat Lung and Liver,” Biochemica et Biophysica Acta, Vol. 582, No. 1, 1979, pp. 741-744.
[15] F. Oberhammer, J. W. Wilson, C. Dive, I. D. Morris, J. A. Hickman, A. E. Wakeling, P. R. Walker and M. Sikorska, “Apoptosis in Epithelial Cells: Cleavages of DNA to 300 and/or 50 kb Fragments Prior to or in the Absence of Inter Nucleosomal Fragmentation,” EMBO Journal, Vol. 12, No. 9, 1993, pp. 3679-3684.
[16] R. J. Anto, M. Venkatraman and D. Karunagaran, “Inhibition of NF ?B sensitizes A431 Cells to Epidermal Growth Factor Induced Apoptosis, Whereas Its Activation by Ectopic Expression of Re1A Confers Resistance,” Journal of Biological Chemistry, Vol. 278, 2003, pp. 25490-25498. doi:10.1074/jbc.M301790200
[17] Y. Y. Soong and P. J. Barlow, “Antioxidant Activity and Phenolic Content of Selected Fruit Seeds,” Food Chemistry, Vol. 88, No. 3, 2004, pp. 411-417. doi:10.1016/j.foodchem.2004.02.003
[18] G. K. Jayaprakasha, R. P. Sigh and K. K. Sakariah, “Antioxidant Activity of Grape Seed (Vitis vinifera) Extracts on Peroxidation Models in Vitro,” Food Chemistry, Vol. 73, No. 3, 2001, pp. 285-290. doi:10.1016/S0308-8146(00)00298-3
[19] Y. F. Li, C. J. Guo, J. J. Yang, J. Y. Wei, J. Xu and S. Cheng, “Evaluation of Antioxidant Properties of Pome-Granate Peel Extract in Comparison with Pomegranate Pulp Extract,” Food Chemistry, Vol. 96, No. 2, 2006, pp. 254-260. doi:10.1016/j.foodchem.2005.02.033
[20] S. Okonogi, C.Duangrat, S. Anuchpreeda, S. Tachakittirungrod and S. Chowwanapoonpohn, “Comparison of Antioxidant Capacities and Cytotoxicities of Certain Fruit Peels,” Food Chemistry, Vol. 103, No. 3, 2007, pp. 839-846. doi:10.1016/j.foodchem.2006.09.034
[21] E. P. Lansky and R. A. Newman “Punica granatum (Pomegranate) and Its Potential for Prevention and Treatment of Inflammation and Cancer,” Journal of Ethnopharmacology, Vol. 109, No. 2, 2007, pp. 177-206. doi:10.1016/j.jep.2006.09.006
[22] M. M. Compton, “A Biochemical Hallmark of Apoptosis: Internucleosomal Degradation of the Genome,” Cancer Metastasis Reviews, Vol. 11, No. 2, 1992, pp. 105-119. doi:10.1007/BF00048058
[23] A. R. Grivell and M. N. Berry, “The Effects of Phosphate-And Substrate-Free Incubation Conditions on Glycolysis in Ehrlich Ascites Tumour Cells,” Biochemica et Biophysica Acta, Vol. 1291, No. 1, 1996, pp. 83-88. doi:10.1016/0304-4165(96)00049-9
[24] T. M. Buttke and P. A. Sandstrom, “Oxidative Stress as a Mediator of Apoptosis,” Immunology Today, Vol. 15, No. 1, 1994, pp. 7-10. doi:10.1016/0167-5699(94)90018-3
[25] T. Arimura, A. Kojima-Yuasa, S. Watanabe, M. Suzuki, D. O. Kennedy and I. Matsui-Yuasa, “Role of Intracellular Reactive Oxygen Species and Mitochondrial Dysfunction in Evening Primrose Extract-Induced Apoptosis in Ehrlich Ascites Tumour Cells,” Chemico-Biological Interaction, Vol. 145, No. 3, 2003, pp. 337-347.
[26] G. S. Salvesen and V. M. Dixit, “Caspases: Intracellular Signaling by Proteolysis,” Cell, Vol. 91, No. 4, 1997, pp. 443-446. doi:10.1016/S0092-8674(00)80430-4
[27] Y. A. Lazebnik, S. H. Kaufmann, S. Desnoyers, G. G. Poirier and W. C. Earnsham, “Cleavage of Poly (ADP-Robose) Polymerase by a Proteinase with Properties Like ICE,” Nature, Vol. 371, 1994, pp. 347-348. doi:10.1038/371346a0
[28] M. Salucci, L. A. Stivala, G. Maiani, R. Bugianesi and V. Vannini, “Flavonoids Uptake and Their Effect on Cell Cycle of Human Colon Adenocarcinoma Cells (CaCO2)” British Journal of Cancer, Vol. 86, 2002, pp. 1645-1651.

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