Isolation of a novel quercetin derivative from Terminalia chebula and RT-PCR-assisted probing to investigate its DNA repair in hepatoma cells

Kallyadan Soumya , Karickal Raman Haridas, Jesna James, Sudhakaran Sudheesh

Abstract


Background and purpose: DNA damage can lead to carcinogenesis if replication proceeds without proper repair. This study focused on the purification of a novel quercetin derivative present in Terminalia chebula fruit and studied its protective role in hepatoma cells due to H2O2-DNA damage. 

Experimental approach: The pure compound obtained from the silica gel column was subjected to structural characterization using spectroscopic techniques. MTT assay was employed to select a non-toxic concentration of the isolated compounds on HepG2 and Chang liver cells. The antigenotoxic property of the compound on HepG2 and Chang liver cells was carried out by alkaline comet assay. Analyses of expression levels of mRNA for two DNA repair enzymes, OGG1 and NEIL1, in HepG2 and Chang liver cells, were carried out using the RT-PCR method.

Findings/Results: The pure compound obtained from the fraction-5 of diethyl ether extract was identified as a novel quercetin derivative and named 7-(but-2-en-1-yloxy)-2-(4(but-2-en-1-yloxy)-3-hydroxyphenyl)-3-(hexa-2,4-dien-1-yloxy)-6-hydroxy-4H-chromen-4-one. This compound recorded modest toxicity at the highest concentration tested (percentage cell viability at 100 µg/mL was 64.71 ± 0.38 for HepG2 and 45.32 ± 0.07 for Chang liver cells). The compound has demonstrated noteworthy protection against H2O2-induced DNA damage in both cell lines. Analyses of mRNA expression levels for enzymes OGG1 and NEIL1 enzymes in HepG2 and Chang liver cells asserted the protective role of the isolated compound against H2O2-induced DNA damage.

Conclusion and implication: The protective effect of a novel quercetin derivative isolated from T. chebula in the hepatoma cells is reported here for the first time.


Keywords


Antigenotoxicity; Chang liver cell; HepG2 cell; NEIL1, OGG1; Quercetin derivative; T. chebula.

Full Text:

PDF

References


Maynard S, Schurman SH, Harboe C, de Souza-Pinto NC, BohrM VA. Base excision repair of oxidative DNA damage and association with cancer and aging. Carcinogenesis. 2009;30(1):2-10. DOI: 10.1093/carcin/bgn250.

Poetsch AR, Boulton SJ, Luscombe NM. Genomic landscape of oxidative DNA damage and repair reveals regioselective protection from mutagenesis. Genome Biol. 2018;19(1):215,1-23.DOI: 0.1186/s13059-018-1582-2.

Lee MR, Kim SH, Cho HJ, Lee KY, Moon AR, Jeong HG, et al. Transcription factors NF-YA regulate the induction of human OGG1 following DNA-alkylating agent methylmethane sulfonate (MMS) treatment. J Biol Chem. 2004;279(11):9857-9866.DOI: 10.1074/jbc.M311132200.

Whitaker AM, Schaich MA, Smith MR, Flynn TS, Freudenthal BD. Base excision repair of oxidative DNA damage: from mechanism to disease. Front Biosci. 2017;22(9):1493-1522.DOI: 10.2741/4555.

Aguiar PHN, Furtado C, Repolês BM, Ribeiro GA, Mendes IC, Peloso EF, et al. Oxidative stress and DNA lesions: the role of 8-oxoguanine lesions in Trypanosoma cruzi cell viability. PLoS Negl Trop Dis. 2013;7(6):e2279,1-13.DOI: 10.1371/journal.pntd.0002279.

Hu J, de Souza-Pinto NC, Haraguchi K, Hogue BA, Jaruga P, Greenberg MM, et al. Repair of formamidopyrimidines in DNA involves different glycosylases: role of the OGG1, NTH1, and NEIL1 enzymes. J Biol Chem. 2005;280(49):40544-40551.DOI: 10.1074/jbc.M508772200.

Saleem A, Husheem M, Härkönen P, Pihlaja K. Inhibition of cancer cell growth by crude extract and the phenolics of Terminalia chebula retz. fruit. J Ethnopharmacol. 2002;81(3):327-336.DOI: 10.1016/S0378-8741(02)00099-5.

Soumya K, Haridas KR, James J, Kumar VBS, Edatt L, Sudheesh S. Study of in vitro antioxidant and DNA damage protection activity of a novel luteolin derivative isolated from Terminalia chebula. J Taibah Univ Sci. 2019;13:755-763.DOI: 10.1080/16583655.2019.1630892.

Soumya K, Jesna J, Sudheesh S. Screening study of three medicinal plants for their antioxidant and cytotoxic activity. Int J Pharm Sci Res. 2018;9(9):3781-3787.DOI: 10.13040/IJPSR.0975-8232.9(9).3781-87.

Malekzadeh F, Ehsanifar H, Shahamat M, Levin M, Colwell RR. Antibacterial activity of black myrobalan (Terminalia chebula Retz) against helicobacter pylori. Int J Antimicrob Agents. 2001;18(1):85-88.DOI: 10.1016/S0924-8579(01)00352-1.

Dutta BK, Rahman I, Das TK. Antifungal activity of Indian plant extracts. Mycoses. 1998;41(11-12):535-536. DOI: 10.1111/j.1439-0507.1998.tb00718x.

Shabrina R, Elya B, Noviani A. Antioxidant activities of fractions from ethyl acetate extracts of Garcinia fruticosa lauterb leaves. Int J Appl Pharm. 2018;10(1):44-50.DOI: 10.22159/ijap.2018.v10s1.10.

Swamy SM, Tan BK. Cytotoxic and immunopotentiating effects of ethanolic extract of Nigella sativa L. seeds. J Ethnopharmacol. 2000;70(1):1-7.DOI: 10.1016/S0378-8741(98)00241-4.

Auddy B, Ferreira M, Blasina F, Lafon L, Arredondo F, Dajas F, et al. Screening of antioxidant activity of three indian medicinal plants, traditionally used for the management of neurodegenerative diseases. J Ethnopharmacol. 2003;84(2-3):131-138.DOI: 10.1016/S0378-8741(02)00322-7.

Kang C, Lee H, Yoo YS, Hah DY, Kim CH, Kim E, et al. Evaluation of oxidative DNA damage using an alkaline single cell gel electrophoresis (SCGE) comet assay, and the protective effects of N-acetylcysteine amide on zearalenone-induced cytotoxicity in chang liver cells. Toxicol Res. 2013;29(1):43-52.DOI: 10.5487/TR.2013.29.1.043.

Leandro LF, Munari CC, Sato VLFL, Alves JM, de Oliveira PF, Mastrocola DFP, et al. Assessment of the genotoxicity and antigenotoxicity of (+)-usnic acid in V79 cells and Swiss mice by the micronucleus and comet assays. Mutat Res. 2013;753(2):101-106.DOI: 10.1016/j.mrgentox.2013.03.006.

Lei YX, Lu Q, Shao C, He CC, Lei ZN, Lian YY. Expression profiles of DNA repair-related genes in rat target organs under subchronic cadmium exposure. Genet. Mol. Res.2015;14(1):515-524. DOI: 10.4238/2015.January.26.5.

Pilařová V, Kuda L, Vlčková HK, Nováková L, Gupta S, Kulkarni M, et al. Carbon dioxide expanded liquid: an effective solvent for the extraction of quercetin from South African medicinal plants. Plant Methods. 2022;18(1):87,1-13.DOI: 10.1186/s13007-022-00919-6.

Silva JP, Gomes AC, Proença F, Coutinho OP. Novel nitrogen compounds enhance protection and repair of oxidative DNA damage in a neuronal cell model: comparison with quercetin. Chem Biol Interact. 2009;181(3):328-337.DOI: 10.1016/j.cbi.2009.07.024.

Ramos AA, Azqueta A, Pereira-Wilson C, Collins AR. Polyphenolic compounds from salvia species protect cellular DNA from oxidation and stimulate DNA repair in cultured human cells. J Agric Food Chem. 2010;58(12):7465-7471. DOI: 10.1021/jf100082p.

García-Rodríguez A, Gosálvez J, Agarwal A, Roy R, Johnston S. DNA damage and repair in human reproductive cells. Int J Mol Sci. 2019;20(1): 31,1-22.DOI: 10.3390/ijms20010031.

Li S, Tan HY, Wang N, Zhang ZJ, Lao L, Wong CW, et al. The role of oxidative stress and antioxidants in liver diseases. Int J Mol Sci. 2015;16(11):26087-26124.DOI: 10.3390/ijms161125942.

Madrigal-Santillán E, Madrigal-Bujaidar E, Álvarez-González I, Sumaya-Martinez MT, Gutiierrez-Salinas J, Bautisa M, et al. Review of natural products with hepatoprotective effects. World J Gastroenterol. 2014;20(40):14787-14804.DOI: 10.3748/wjg.v20.i40.14787.

Srivastava R, Srivastava P. Hepatotoxicity and the role of some herbal hepatoprotective plants in present scenario. Global J Dig Dis. 2018;3:1-4.DOI: 10.4172/2472-1891.100034.

Linares V, Alonso V, Albina ML, Bellés M, Sirvent JJ, Domingo JL, et al. Lipid peroxidation and antioxidant status in kidney and liver of rats treated with sulfasalazine. J Toxicol. 2009;256(3):152-156.DOI: 10.1016/j.tox.2008.11.010.

Kilani-Jaziri S, Bhouri W, Skandrani I, Limem I, Chekir-Ghedira L, Ghedira K. Phytochemical, antimicrobial, antioxidant and antigenotoxic potentials of Cyperus rotundus extracts. S. Afr. J. Bot. 2011;77(3):767-776.DOI: 10.1016/j.sajb.2011.03.015.

Prajitha V, Thoppi J.E. Genotoxic and antigenotoxic potential of the aqueous leaf extracts of Amaranthus spinosus Linn. using Allium cepa assay. S Afr J Bot 2016;102:18-25.DOI: 10.1016/j.sajb.2015.06.018.

Prieto AM, dos Santos AG, Oliveira APS, Cavalheiro AJ, Silva DHS, Bolzani VS, et al. Assessment of the chemopreventive effect of casearin B, a clerodane diterpene extracted from Casearia sylvestris (Salicaceae). Food Chem Toxicol. 2013;53:153-159.DOI: 10.1016/j.fct.2012.11.029.

Benhusein GM, Mutch E, Aburawi S, Williams FM. Genotoxic effect induced by hydrogen peroxide in human hepatoma cells using comet assay. Libyan J Med. 2010;5:1-6.DOI: 10.3402/ljm.v5i0.4637.

Ismail IH, Nyström S, Nygren J, Hammarsten O. Activation of ataxia telangiectasia mutated by DNA strand break-inducing agents correlates closely with the number of DNA double strand breaks. J Biol Chem. 2005;280(6):4649-4655.DOI: 10.1074/jbc.M411588200.

Kim KC, Lee IK, Kang KA, Cha JW, Cho SJ, Na SY, et al. 7,8-Dihydroxyflavone suppresses oxidative stress-induced base modification in DNA via induction of the repair enzyme 8-oxoguanine DNA glycosylase-1. Biomed Res Int. 2013;2013:863720,1-10.DOI: 10.1155/2013/863720.

Kang JS, Choi IW, Han MH, Kim GY, Hong SH, Park C, et al. The cytoprotective effects of 7,8-dihydroxyflavone against oxidative stress are mediated by the upregulation of Nrf2-dependent HO-1 expression through the activation of the PI3K/Akt and ERK pathways in C2C12 myoblasts. Int J Mol Med. 2015;36(2): 501-510.DOI: 10.3892/ijmm.2015.2256.

Silva JP, Gomes AC, Coutinho OP. Oxidative DNA damage protection and repair by polyphenolic compounds in PC12 cells. Eur J Pharmacol. 2008;601(1-3):50-60. DOI: 10.1016/j.ejphar.2008.10.046.


Refbacks

  • There are currently no refbacks.


Creative Commons LicenseThis work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License which allows users to read, copy, distribute and make derivative works for non-commercial purposes from the material, as long as the author of the original work is cited properly.