Reveal Cytotoxicity and Antigenotoxicity of Piper nigrum L. Ethanolic Extract and its Combination with Doxorubicin on CHO-K1 Cells

Nur Fitra Sari, Beni Lestari, Dian Saputri, Anisa Fauzia Ahsani, Ragil Anang Santoso, Ediati Sasmito, Edy Meiyanto

Abstract


Black pepper (Piper nigrum L.), one of the most popular Indonesian spices has been reported to possess various therapeutic effects. The aim of this study is to evaluate the cytotoxicity and antigenotoxicity of black pepper ethanolic extract (BPE) and its combination with doxorubicin (Dox) on CHO-K1 cells. Based on thin layer chromatographyanalysis, BPE contained piperine.Under MTT assay, BPE showed cytotoxic effect with the IC50 value of 68 μg/mL and performed synergism in combination with Dox. In vitro micronucleus test using Giemsa staining revealed that BPE did not cause morphological changes qualitatively on CHO-K1 cells at concentration of 8.5 μg/mL, whereas using flow cytometry analysis showed that BPE could decrease the number of micronucleus (MN) formation induced by doxorubicin. In addition, BPE reduced the ROS level on the CHO-K1 cells which observed by reactive oxygen species (ROS) intracellular assay. The decrease in ROS level indicated that the antioxidant activity of BPE contribute to the antigenotoxicity. Furthermore, molecular docking performed that piperine interacted with DNA Topoisomerase II with docking score of -80.68. Overall,BPE performed cytotoxic effect in single treatment, increased the cytotoxicity and reduced the genotoxicity of doxorubicin. Thus, BPE has potential to be developed further as co-chemotherapeutic and antigenotoxic agent.

Keywords: Cytotoxic, genotoxic, Piper nigrum L., CHO-K1, micronucleus


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Ahmad, N., Fazal, H., Abbasi, B. H., Farooq, S., Ali, M. and Khan, M. A., 2012, Biological role of Piper nigrum L. (Black pepper): A review, Asian Pac. J. Trop. Biomed., 2(3), S1945-S1953. CrossRef

Andrews, A.W., Fornwald, J.A. and Lijinsky, W., 1980, Nitrosation and Mutagenicity of Some Amine Drugs, Toxicol. Appl. Pharmacol., 52(2), 237–244. CrossRef

Avlasevich, S.L., Bryce, S.M., Cairns, S.E. and Dertinger, S.D., 2006, In Vitro Micronucleus Scoring by Flow Cytometry: Differential Staining of Micronuclei Versus Apoptotic and Necrotic Chromatin Enhances Assay Reliability, Environ. Mol. Mutagen., 47(1), 56-66. CrossRef

Bajad, S., Bedi, K.L., Singla, A.K. and Johri, R.K., 2001, Antidiarrhoeal Activity of Piperine in Mice, Planta Medica, 67(3), 284-287. CrossRef

Balakrishnan, S., Vellaichamy, L., Menon, V.P. and Manoharan, S., 2008, Antigenotoxic Effects of Curcumin and Piperine Alone or in Combination Against 7,12- dimethylbenz(a)anthracene Induced Genotoxicity in Bone Marrow of Golden Syrian Hamsters, Toxicol. Mech. Methods, 18(9), 691–696. CrossRef

Bruton, L., Lazo, J.S. and Parker, K.L., 2005, Goodman & Gilman’s The Pharmacological Basis of Therapeutics, 11th Edition, Pennsylvania: McGrawHill.

Bryce, S.M., Shi, J., Nicolette, J., Diehl, M., Sonders, P., Avlasevich, S. and Dertinger, S.D., 2010, High Content Flow Cytometric Micronucleus Scoring Method is Applicable to Attachment Cell Lines, Environ. Mol. Mutagen., 51(3), 260-266. CrossRef

Carvalho, C., Santos, R.X., Cardoso, S., Correia, S., Oliveira, P.J., Santos, M.S., et al, 2009, Doxorubicin: The Good, the Bad and the Ugly Effect, Curr. Med. Chem., 16(25), 3267-3285. CrossRef

Damanhouri, Z. A. and Ahmad, A., 2014, A Review on Therapeutic Potential of Piper nigrum L. (Black Pepper): The King of Spices, Med Aromat Plant, 3(3), 1-6. CrossRef

Fenech, M., 2000, The in Vitro Micronucleus Technique, Mutat. Res. Fundam. Mol. Mech. Mutagen., 455(1), 81-95. CrossRef

Gülçin, İ., 2005, The Antioxidant and Radical Scavenging Activities of Black Pepper (Piper nigrum) Seeds, Int. J. Food Sci. Nutr., 56(7), 491-499. CrossRef

Karekar, V.R., Mujumdar, A.M., Joshi, S.S., Dhuley, J., Shinde, S.L. and Ghaskadbi, S., 1996, Assessment of Genotoxic Effect of Piperine using Salmonella typhimurium and Somatic and Germ Cells of Swiss Albino Mice, Arzneimittel-Forschung/Drug Res., 46(10), 972–975.

Khan, M., Maryam, A., Mehmood, T., Zhang, Y. and Ma, T., 2015, Enhancing Activity of Anticancer Drugs in Multidrug Resistant Tumors by Modulating P-Glycoprotein through Dietary Nutraceuticals, Asian Pac. J. Cancer Prev., 16(16), 6831-6839. CrossRef

Li, S., Wang, C., Wang, M., Li, W., Matsumoto, K. and Tang, Y., 2007. Antidepressant Like Effects of Piperine in Chronic Mild Stress Treated Mice and Its Possible Mechanisms, Life Sci., 80(15), 1373-1381. CrossRef

Lu, J.J., Bao, J.L., Chen, X.P., Huang, M. and Wang, Y.T., 2012, Alkaloids Isolated from Natural Herbs as The Anticancer Agents, Evid. Based Complement. Alternat. Med., 2012, 485042, doi: 10.1155/2012/485042. CrossRef

Manoharan, S., Balakrishnan, S., Menon, V., Alias, L. and Reena, A., 2009, Chemopreventive Efficacy of Curcumin and Piperine during 7,12-dimethylbenz[a]anthracene-induced Hamster Buccal Pouch Carcinogenesis, Singapore Med. J., 50(2), 139-146.

Meghwal, Murlidhar and Goswami., T.K., 2013, Piper nigrum and Piperine: An Update, Phytother. Res., 27(8), 1121-1130. CrossRef

Mosmann, T., 1983, Rapid Colorimetric Assay for cellular growth and Survival: Application to Proliferation and Cytotoxicity Assays, J. Immunol. Methods., 65(1-2), 65-59. CrossRef

Muralidhara and Narasimhamurthy, K., 1990, Lack of Genotoxic Effects of Piperine, (The Active Principle of Black Pepper) in Albino Mice, J. Food Saf., 11(1), 39–48. CrossRef

Paarakh, P.M., Sreeram, D.C., Shruthi, S.D. and Ganapathy, P.S., 2015, In Vitro Cytotoxic and in Silico Activity of Piperine Isolated from Piper nigrum fruits Linn, In Silico Pharmacology, 3(1), 9. CrossRef

Parganiha, R., Verma, S., Chandrakar, S., Pal, S., Sawarkar, H.A. and Kashyap, P., 2011, In Vitro Anti-asthmatic Activity of Fruit Extract of Piper nigrum (Piperaceae), Inter. J. Herbal Drug Res., 1(1), 15-18.

Prayong, P., Barusrux, S. and Weerapreeyakul, N., 2008, Cytotoxic Activity Screening of Some Indigenous Thai Plants, Fitoterapia, 79(7), 598-601. CrossRef

Rasheed, M., Afshan, F., Tariq, R.M., Siddiqui, B.S., Gulzar, T., Mahmood, A., et al, 2005, Phytochemical Studies on The Seed Extract of Piper nigrum Linn., Nat. Prod. Res., 19(7), 703–712.

Sehgal, A., Kumar, M., Jain, M. and Dhawan, D.K., 2013, Modulatory Effect of Curcurmin in Conjunction with Piperine on benzo(A)pyrene-mediated DNA Adducts and Biotransformation Enzymes, Nutr. Cancer, 65(6), 885–890. CrossRef

Selvendiran, K., Singh, J.P.V., Krishnan, K.B. and Sakthisekaran, D., 2003, Cytoprotective Effect of Piperine against Benzo[a]pyrene Induced Lung Cancer with Reference to Lipid Peroxidation and Antioxidant System in Swiss Albino Mice, Fitoterapia, 74(1-2), 109–115. CrossRef

Singh, J., Reen, R.K. and Wiebel, F.J., 1994, Piperine, A Major Ingredient of Black and Long Peppers, Protects Against AFB1-induced Cytotoxicity and Micronuclei Formation in H4IIEC3 Rat Hepatoma Cells, Cancer Lett., 86(2), 195–200. CrossRef

Taqvi, S.I., Shah, A.J. and Gilani, A.H., 2008, Blood Pressure Lowering and Vasomodulator Effects of Piperine, J. Cardiovasc. Pharmacol., 52(5), 452-458. CrossRef

Thiel, A., Buskens, C., Woehrle, T., Etheve, S., Schoenmakers, A., Fehr, M., et al., 2014, Black Pepper Constituent Piperine: Genotoxicity Studies in Vitro and in Vivo, Food Chem. Toxicol., 66, 350-357. CrossRef

Vellaichamy, L., Balakrishnan, S., Panjamurthy, K., Manoharan, S. and Alias, L.M., 2009, Chemopreventive Potential of Piperine in 7,12-dimethylbenz[a]anthracene-induced Skin Carcinogenesis in Swiss Albino Mice, Environ. Toxicol. Pharmacol., 28(1), 11–18. CrossRef




DOI: http://dx.doi.org/10.14499/indonesianjcanchemoprev8iss3pp110-119

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