Identification of polyphenols in leaf extracts of Lawsonia inermis L. with antioxidant, antigenotoxic and antiproliferative potential
DOI:
https://doi.org/10.22377/ijgp.v8i1.350Abstract
Background: Keeping in view the importance of natural plant products, bioactivity‑guided isolation of phytoconstituents was attempted from Lawsonia inermis L. Materials and Methods: Various polar and non‑polar extract/fractions were isolated and evaluated for the genotoxic and antigenotoxic potential against mutagens viz., 4‑nitroquinoline‑1‑oxide and nitrofurantoin in SOS chromotest using Escherichia coli PQ37 tester strain and against H2O2 in DNA protection assay. In order to decipher mode of action, antioxidant activity was evaluated using different assays. The extract/fractions were also investigated for their antiproliferative potential against PC‑3 and Colo205 cancer cell lines. Major polyphenolic constituents were identified using high-performance liquid chromatography (HPLC) technique.
Results: All the extract/fractions except Hex‑Li significantly decreased the SOS‑inducing potency of both the mutagens. In various in vitro assays, the polar fractions exhibited marked antioxidant activity. HPLC analysis of the extract/fractions showed that gallic acid, catechin, chlorogenic acid, epicatechin, rutin, ellagic acid, quercetin and kaempferol are the major constituents. The marked antigenotoxic activity of extract/fractions may be attributed to the polyphenolic compounds. The fractions viz., Hex‑Li and CHCl3‑LI were found to be highly cytotoxic against Colo‑205 cancer cell line. Conclusions: This is the first report of antigenotoxic and anticancer activities of Lawsonia inermis L. phytoconstituents. The antiradical potency of these extract/fractions may be responsible for the above activities.
Key words: 4‑Nitroquinoline‑1‑oxide, antioxidant activity, cancer cell lines, nitrofurantoin, SOS chromotest
Downloads
References
Steele VE. Current mechanistic approaches to the chemoprevention
of cancer. J Biochem Mol Biol 2003;36:78‑81.
Manson MM. Cancer prevention: The potential for diet to modulate
molecular signaling. Trends Mol Med 2003;9:11‑8.
Ames BN, Shigenaga MK, Gold LS. DNA lesions, inducible DNA
repair, and cell division: Three key factors in mutagenesis and
carcinogenesis. Environ Health Perspect 1993;101:35‑44.
Liu RH, Hotchkiss JH. Potential genotoxicity of chronically
elevated nitric oxide: A review. Mutat Res 1995;339:73‑89.
Sporn MB. Approaches to prevention of epithelial cancer during
the preneoplastic period. Cancer Res 1976;36:2699‑702.
Surh YJ. Cancer chemoprevention with dietary phytochemicals.
Nat Rev Cancer 2003;3:768‑80.
Bode AM, Dong Z. Targeting signal transduction pathways by
chemopreventive agents. Mutat Res 2004;555:33‑51.
Chun KS, Surh YJ. Signal transduction pathways regulating Cox‑2
expression: Potential molecular targets for chemoprevention.
Biochem Pharmacol 2004;68:1089‑100.
Bringmann G, Münchbach M, Feineis D, Faulhaber K,
Ihmels H. Studies on single‑strand scissions to cell‑free
p l a s m i d D N A b y t h e d o p a m i n e r g i c n e u r o t o x i n
‘TaClo’ (1‑trichloromethyl‑1,2,3,4‑tetrahydro‑beta‑carboline).
Neurosci Lett 2001;304:41‑4.
Halliwell B, Gutteridge JMC. In: Oxidative stress, in Free Radicals
in Biology and Medicine. 3rd ed. Halliwell B., Gutteridge J.M.C.,
editors. Oxford University Press; New York: 1999. pp. 246–350.
Valko M, Rhodes CJ, Moncol J, Izakovic M, Mazur M. Free radicals,
metals and antioxidants in oxidative stress‑induced cancer. Chem
Biol Interac 2006;160:1‑40.
Murphy MP. How mitochondria produce reactive oxygen species.
Biochem J 2009;417:1‑13.
Niciforovic N, Mihailovic V, Maskovic P, Solujic S, Stojkovic A,
Muratspahic DP. Antioxidant activity of selected plant species;
potential new sources of natural antioxidants. Food Chem Toxicol
;48:3125‑30.
Conforti F, Sosa S, Marrelli M, Menichini F, Statti GA, Uzunov D,
et al. In vivo anti‑inflammatory and in vitro antioxidant activities of
Mediterranean dietary plants. J Ethnopharmacol 2008;116:144‑51.
Newman DJ, Cragg GM, Snader KM. Natural products as
sources of new drugs over the period 1981‑2002. J Nat Prod
;66:1022‑37.
Dorai T, Aggarwal BB. Role of chemopreventive agents in cancer
therapy. Cancer Lett 2004;215:129‑40.
Chetty KM. Flowering plants of Chittoor. 1st ed. Andhra Pradesh;
p. 132.
Chopra RN, Nayer SL, Chopra IC. Glossary of Indian medicinal
plants. New Delhi: CSIR Publications; 1956. p. 151.
Reddy KR. Folk medicine from Chittoor district Andhra Pradesh,
India used in the treatment of jaundice. Int J Crude Drug Res
;26:137‑40.
Yu L, Haley S, Perrat J, Harris M, Wilson J, Qian M. Free radical
scavenging properties of wheat extracts. J Agric Food Chem
;50:1619‑24.
Kim D, Jeong S, Lee CY. Antioxidant capacity of phenolic
phytochemicals from various cultivars of plums. Food Chem
;81:321‑6.
Quillardet P, Hofnung M. The SOS Chromotest, a colorimetric
bacterial assay for genotoxins: Procedures. Mutat Res
;147:65‑78.
Kevekordes S, Merch‑Sundermann V, Burghaus CM, Spielberg J,
Schmeiser H, Arlt VM, et al. SOS induction of selected naturally
occurring substances in Escerichia coli (SOS chromotest). Mutat
Res 1998;445:81‑91.
Lee JC, Kim HR, Kim J, Jang YS. Antioxidant property of an
ethanol extract of the stem of Opuntia ficus‑indica var. Saboten.
J Agric Food Chem 2002;50:6490‑6.
Marcocci L, Maguire JJ, Droy‑Lefaix MT, Packer L. The nitric
oxide‑scavenging properties of Ginkgo biloba extract EGb 761.
Biochem Biophys Res Commun 1994;201:748‑55.
Re R, Pellegrini N, Proteggente A, Pannala A, Yang M,
Rice‑Evans C. Antioxidant activity applying an improved
ABTS radical cation decolorization assay. Free Radic Biol Med
;26:1231‑7.
Blois MS. Antioxidant determinations by the use of a stable free
radical. Nature 1958;18:1199‑200.
Oyaizu M. Studies on product of browning reaction prepared
from glucose amine. Jpn J Nutr 1986;44:307‑15.
Nishikimi M, Appaji N, Yagi K. The occurrence of superoxide
anion in the reaction of reduced phenazine methosulfate and
molecular oxygen. Biochem Biophys Res Commun 1972;46:849‑54.
Prieto P, Pineda M, Aguilar M. Spectrophotometric
quantitation of antioxidant capacity through the formation of
a phosphomolybdenum complex: Specific application to the
determination of vitamin E1. Anal Biochem 1999;269:337‑41.
Mickisch G, Fajta S, Keilhauer G, Schlick E, Tschada R, Alken P.
Chemosensitivity testing of primary human renal cell carcinoma
by a tetrazolium based microculture assay (MTT). Urol Res
;18:131‑6.
Storz P. Reactive oxygen species in tumor progression. Front
Biosci 2005;10:1881‑96.
Radtke F, Clevers H. Self‑renewal and cancer of the gut: Two
sides of a coin. Science 2005;307:1904‑9.
Slupphaug G, Kavli B, Krokan HE. The interacting pathways
for prevention and repair of oxidative DNA damage. Mutat Res
;531:231‑51.
Nakahara W, Fukuoka F, Sugimura T. Carcinogenic action of
‑nitroquinoline‑N‑oxide. Gan 1957;48:129‑37.
Kitano M. Host genes controlling the susceptibility and
resistance to squamous cell carcinoma of the tongue in a rat
model. Pathol Int 2000;50:353‑62.
Tada M. Seryl‑tRNA synthetase and activation of the carcinogen
‑nitroguinoline 1‑oxide. Nature 1975;255:510‑2.
Kohda K, Kawazoe Y, Minoura Y, Tada M. Separation and
identification of N4‑(guanosin‑7‑yl)‑4‑aminoquinoline 1‑oxide, a
novel nucleic acid adduct of carcinogen 4‑nitroquinoline 1‑oxide.
Carcinogenesis 1991;12:1523‑5.
B a i l l e u l B , D a u b e r s i e s P , G a l i e g u e ‑ Z o u i t i n a S ,
Loucheux‑Lefebvre MH. Molecular basis of 4‑nitroquinoline
‑oxide carcinogenesis. Jpn J Cancer Res 1989;80:691‑7.
Nagao M, Sugimura T. Molecular biology of the carcinogen,
‑nitroquinoline 1‑oxide. Adv Cancer Res 1976;23:131‑69.
Miao ZH, Rao VA, Agama K, Antony S, Kohn KW, Pommier Y.
‑Nitroquinoline‑1‑Oxide Induces the Formation of Cellular
Topoisomerase I‑DNA Cleavage Complexes. Cancer Res
;66:6540‑5.
Huang MH, Tai HM, Wang BS, Chang LW. Inhibitory effects of
water extract of flos inulae on mutation and tyrosinase. Food
Chem 2013;139:1015‑20.
Kang Z-C, Yen M-T, Chiu C-K, Wu H-C, Huang S-L, Tai S-P,
et al. The inhibitory effects of aqueous extract from guava twigs,
Psidium guajava L., on mutation and oxidative damage. J Chem
; doi:10.1155/2013/561905.
Tanaka T, Kojima T, Kawamori T, Wang A, Suzui M, Okamoto K,
et al. Inhibition of 4‑nitroquinoline‑1‑oxide‑induced rat tongue
carcinogenesis by the naturally occurring plant phenolics
caffeic, ellagic, chlorogenic and ferulic acids. Carcinogenesis
;14:1321‑5.
Tu Y, McCalla DR. Effect of activated nitrofurans on DNA.
Biochim Biophys Acta 1975;402:142‑9.
Bryant B, Knights K. Pharmacology for health professionals. In:
rd ed. Unit‑14 Drugs Affecting Microorganism; 2010. p. 868.
Bhouri W, Sghaier MB, Kilani S, Bouhlel I, Dijoux‑Franca MG,
Ghedira K, et al. Evaluation of antioxidant and antigenotoxic
a c t i v i t y o f t w o f l a v o n o i d s f r o m Rhamnus alaternus
L. (Rhamnaceae): kaempferol 3‑O‑β‑isorhamninoside and
rhamnocitrin 3‑O‑β‑isorhamninoside. Food Chem Toxicol
;49:1167‑73.
Kada T, Inoue T, Ohta T, Shirasu Y. Antimutagens and their
modes of action. Basic Life Sci 1986;39:181‑96.
Kaur S, Grover IS, Kumar S. Antimutagenic potential of
ellagic acid isolated from Terminalia arjuna. Indian J Exp Biol
;35:478‑82.
Festa F, Aglitti T, Duranti G, Ricordy R, Perticone P, Cozzi R.
Strong antioxidant activity of ellagic acid in mammalian
cells in vitro revealed by the comet assay. Anticancer Res
;21:3903‑8.
Talcott ST, Lee JH. Ellagic acid and flavonoid antioxidant
content of muscadine wine and juice. J Agric Food Chem
;50:3186‑92.
Seeram NP, Adams LS, Henning SM. In vitro antiproliferative,
apoptotic and antioxidant activities of punicalagin, ellagic
acid and a total pomegranate tannin extract are enhanced in
combination with other polyphenols as found in pomegranate
juice. J Nutr Biochem 2005;16:360‑7.
Han DH, Lee MJ, Kim JH. Antioxidant and apoptosis inducing
activities of ellagic acid. Anticancer Res 2006;26:3601‑6.
Soong YY, Barlow PJ. Quantification of gallic acid and
ellagic acid from longan (Dimocarpus longan Lour.) seed
and mango (Mangifera indica L.) kernel and their effects on
antioxidant activity. Food Chem 2006;97:524‑30.
Torel J, Cillard J, Cillard P. Antioxidant activity of flavonoids and
reactivity with peroxy radical. Phytochemistry 1986;25:383‑5.
Chu YH, Chang CL, Hsu HF. Flavonoid content of several
vegetables and their antioxidant activity. J Sci Food Agric
;80:561‑6.
Singh R, Singh B, Singh S, Kumar N, Kumar S, Arora S. Anti‑freeradical activities of kaempferol isolated from Acacia nilotica (L.)
Willd. E ×. Del. Toxicol In vitro 2008;22:1965‑70.
Choi JS, Park KY, Moon SH, Rhee SH, Young HS. Antimutagenic
effect of plant flavonoids in the Salmonella assay system. Arch
Pharm Res 1994;17:71‑5.
Chen ZY, Chan PT, Ho KY, Fung KP, Wang J. Antioxidant activity
of natural flavonoids is governed by number and location of their
aromatic hydroxyl groups. Chem Phys Lipids 1996;79:157‑63.
Kweon MH, Hwang HJ, Sung HC. Identification and
antioxidant activity of novel chlorogenic acid derivatives from
bamboo (Phyllostachys edulis). J Agric Food Chem 2001;49:4646‑55.
Sato Y, Itagaki S, Kurokawa T, Ogura J, Kobayashi M, Hirano T,
et al. In vitro and in vivo antioxidant properties of chlorogenic acid
and caffeic acid. Int J Pharm 2011;403:136‑8.
Golumbic C, Mattill HA. The antioxidant properties of gallic acid
and allied compounds. J Am Oil Chem Soc 1942;19:144‑5.
Aruoma OI, Murcia A, Butler J, Halliwell B. Evaluation of the
antioxidant and prooxidant actions of gallic acid and its derivatives.
J Agric Food Chem 1993;41:1880‑5.
Brand‑Williams W, Cuvelier ME, Berset C. Use of a free radical
method to evaluate antioxidant activity. LWT‑Food Sci Technol
;28:25‑30.
Yen GC, Duh PD, Tsai HL. Antioxidant and pro‑oxidant properties
of ascorbic acid and gallic acid. Food Chem 2002;79:307‑13.
Abdelwahed A, Bouhlel I, Skandrani I, Valenti K, Kadri M,
Guiraud P, et al. Study of antimutagenic and antioxidant activities
of gallic acid and 1, 4, 6‑pentagalloylglucose from Pistacia lentiscus
confirmation by microarray expression profiling. Chem Biol
Interact 2007;165:1‑13.
Scalzo RL. Measurement of free radical scavenging activity of gallic
acid and unusual antioxidants as sugars and hydroxyacids. Elec
J Env Agricult Food Chem 2010;9:1360‑71.
Taubert D, Breitenbach T, Lazar A, Censarek P, Harlfinger S,
Berkels R, et al. Reaction rate constants of superoxide scavenging
by plant antioxidants. Free Radic Biol Med 2003;35:1599‑607.
Adhami VM, Malik A, Zaman N, Sarfaraz S, Siddiqui IA, Syed DN,
et al. Combined inhibitory effects of green tea polyphenols and
selective cyclooxygenase‑2 inhibitors on the growth of human
prostate cancer cells both in vitro and in vivo. Clin Cancer Res
;13:1611‑9.
Plochmann K, Korte G, Koutsilieri E, Richling E, Riederer P,
Rethwilm A, et al. Structure‑activity relationships of
flavonoid‑induced cytotoxicity on human leukemia cells. Arch
Biochem Biophys 2007;460:1‑9.
Middleton E Jr, Kandaswami C, Theoharides TC. The effect of
plant flavonoids on mammalian cell: Implication for inflammation,
heart disease and cancer. Pharmacol Rev 2000;52:673‑751.
Sharma V, Joseph C, Ghosh S, Agarwal A, Mishra MK, Sen E.
Kaempferol induces apoptosis in glioblastoma cells through
oxidative stress. Mol Cancer Ther 2007;6:2544‑53.
Luo H, Daddysman MK, Rankin GO, Jiang BH, Chen YC.
Kaempferol enhances cisplatin’s effect on ovarian cancer cells
through promoting apoptosis caused by down regulation of cMyc.
Cancer Cell Int 2010;10:16.
Bosetti C, Bravi F, Talamini R, Parpinel M, Gnagnarella P, Negri E,
et al. Flavonoids and prostate cancer risk: A study in Italy. Nutr
Cancer 2006;56:123‑7.
Theodoratou E, Kyle J, Cetnarskyj R, Farrington SM, Tenesa A,
Barnetson R, et al. Dietary flavonoids and the risk of colorectal
cancer. Cancer Epidemiol Biomarkers Prev 2007;16:684‑93.
Bosetti C, Rossi M, McLaughlin JK, Negri E, Talamini R, Lagiou P,
et al. Flavonoids and the risk of renal cell carcinoma. Cancer
Epidemiol Biomarkers Prev 2007;16:98‑101.
Mertens‑Talcott SU, Talcott ST, Percival SS. Low concentrations of
quercetin and ellagic acid synergistically influence proliferation,
cytotoxicity and apoptosis in MOLT‑4 human leukemia cells.
J Nutr 2003;133:2669‑74.
