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Lawsonia Inermis A tica nal eu yt c ic a a m A r a c t Babili et al., Pharmaceut Anal Acta 2013, 4:1 h a P DOI: 10.4172/2153-2435.1000203 ISSN: 2153-2435 Pharmaceutica Analytica Acta Research Article Open Access Lawsonia Inermis: Its Anatomy and its Antimalarial, Antioxidant and Human Breast Cancer Cells MCF7 Activities Fatiha El Babili1,3*, Alex Valentin2 and Christian Chatelain1 1Faculté des Sciences Pharmaceutiques de Toulouse-Laboratoire de Botanique-Mycologie-Faculté de Pharmacie, 35 ch. des Maraîchers - F- 31062 Toulouse, France 2Laboratoire de Pharmacochimie des Substances Naturelles et Pharmacophores Redox, UMR 152, UPS, Université de Toulouse, 118 Route de Narbonne, F-31062 Toulouse Cedex 9, France 3Laboratoire des Interactions Moléculaires et Réactivité Chimique et Photochimique UMR CNRS 5623, Université de Toulouse, 118 Route de Narbonne, F-31062 Toulouse Cedex 9, France Abstract Anatomical features were highlighted to improve its identification, because although henna has some benefits, it has a wide spectrum of adverse effects from contact dermatitis to severe angioneurotic oedema and haemolysis. Serial extractions in petroleum ether, ethyl acetate, ethanol and water were performed on leaves of Lawsonia inermis and were studied. In these extracts, different chemical families were measured such as polyphenols (Gallic acid equivalent 71.7-129.6 g/Kg), tannins (Catechin equivalent 31.3-477.9 g/Kg), antocyanins (Cyanidin equivalent 0.75-5.48 mg/Kg) and flavonoids (Quercetin equivalent 16.2-85.6 g/Kg). This present study reports ABTS/DPPH assay and antimalarial activity. We also checked activity against human breast cancer cells MCF7. The best extract with an antioxidant activity (IC50=6.9 ± 0.1 mg/L) was obtained by ethanol. We found an antimalarial activity of the petroleum ether extract (27 mg/L) of henna. Henna extracts showed activity against human breast cancer cells (MCF7) with the ethyl acetate extract (27 mg/L) and petroleum extract (22 mg/L). Keywords: Lawsonia inermis; Antioxidant activity; Antimalarial Apigenin-7-glucoside, apigenin-4’-glucoside, luteolin-7-glucoside, activity; Cytotoxicity MCF7 and luteolin-3’-glucoside were isolated from leaf extracts of L. innermis. Its methanol extract yielded β-sitosterol, stigmasterol and Introduction 1,2-dihydroxy-4-glucosyloxynaphthalene. The leaves of L. inermis Lawsonia inermis L. (= L. alba Lamk.), belonging to the Lythraceae yielded luteolin, acacetin-7-O-glucoside, glucoside of β-sitosterol family, is a very widespread medicinal plant and natural dye in the [6]. Two pentacyclic triterpenes isolated from the bark of L. inermis world. This herb which has interesting dyeing properties was used (henna) were identified as β3 ,30-dihydroxylup-20(29)-ene (hennadiol) traditionally for centuries in Asia, in Africa for dyeing hair and painting and (20S)-3β,30-dihydroxylupane [6]. skin and nails. Since 1890, it has been widely distributed to Europe and In 1992, a sterol, namely lawsaritol, was isolated from the roots nowadays available in worldwide market as ingredient in many hair of L. inermis and elucidated as 24β-ethylcholest-4-en-3β-ol and from dyes and hair-care products. Lawsonia. inermis, also called “henna”, the stem bark of L. inermis 3-methylnonacosan-1-ol [7]. The methanol contains many kinds of chemical compounds, such as coumarin, extract of the defatted L. inermis seeds has led to the isolation of two quinones, flavonoids etc. and has stronger physiological activity. Due triterpenoids lawnermis acid and its methyl ester [8]. Two pentacyclic to its natural and harmless characters, the Henna is used as a kind triterpenoids have been isolated from the aerial parts of Lawsonia alba. of natural dye, and is used as a raw material for natural hair dyes. In The structures of the new constituents, named as lawsonic acid (I) and addition, Henna can be used as dyes for textile and tattoo material. The lawsonin (II) have been elucidated as 3β-E-ferulyloxy-lup-20(29)- medicinal value of L. inermis is very high and it is used for treatments en-28-oic acid and 3β-E-ferulyloxy-urs-11-en-13β-ol respectively, of headache, jaundice, leprosy [1]. In addition, Henna was shown as an through spectroscopic studies [9]. unusual cause of children suicide [2]. The biological properties of henna were also studied as follows: The phytochemistry of henna was largely studied and revealed interesting information. Already in 1920 the dye principle was known. Crude ethanol extract of Lawsonia inermis L. produced anti- Lawsone, C10H6O3, the colouring matter contained in henna leaves, is fixed well by wool, silk and tenaciously by the skin [3]. In 1973, four fractions with antibacterial activity were isolated by thin-layer *Corresponding author: Fatiha El Babili, Laboratoire de Botanique-Mycologie, chromatography of the ethanol extract of Lawsonia inermis leaves. 3 Faculté de Pharmacie, Université de Toulouse, 118 Route de Narbonne, F-31062 of which were identified as gallic acid [149-91-7], lawsone (2-hydroxy- Toulouse Cedex 9, France, Tel: +33562259817; E-mail: [email protected] 1,4-naphthoquinone)(I) [83-72-7], and 1,4-naphthoquinone [130-15- Received November 26, 2012; Accepted January 23, 2013; Published January 4] [4]. 25, 2013 Then in 1976, a coumarin, lacoumarin, was isolated from L. in- Citation: Babili FE, Valentin A, Chatelain C (2013) Lawsonia Inermis: Its Anatomy armis. It was then the turn of xanthones; two xanthones isolated from and its Antimalarial, Antioxidant and Human Breast Cancer Cells MCF7 Activities. Pharmaceut Anal Acta 4: 203. doi:10.4172/2153-2435.1000203 L. inermis were characterized as 1,3-dihydroxy-6,7-dimethoxyxantho- ne and 1-hydroxy-3,6-diacetoxy-7-methoxyxanthone and named lax- Copyright: © 2013 Babili FE, et al. This is an open-access article distributed under anthone I and II, respectively. Another xanthone, named laxanthone the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and III was identified from chemical and spectral data [5]. source are credited. Pharmaceut Anal Acta ISSN: 2153-2435 PAA, an open access journal Volume 4 • Issue 1 • 1000203 Citation: Babili FE, Valentin A, Chatelain C (2013) Lawsonia Inermis: Its Anatomy and its Antimalarial, Antioxidant and Human Breast Cancer Cells MCF7 Activities. Pharmaceut Anal Acta 4: 203. doi:10.4172/2153-2435.1000203 Page 2 of 6 inflammatory, analgesic, molluscicidal, antibacterial and antipyretic Total amount of phenolic compounds effects. The ethanolic extract of Lawsonia inermis leaves and lawsone The total phenolic amount of each extract was determined by the tested for trypsin inhibitory activity showed an IC50 value of 64.87 and 48.6 microg/ml, respectively [10]. Its ethanol leaf extract was Folin-Ciocalteu method [16]. A diluted solution of each extract (0.5 active against a wide range of microbes, such as Staphylococcus, mL) was mixed with Folin Ciocalteu reagent (0.2 N, 2.5 mL). This Streptococcus, Brucella and Salmonella, but was inactive against mixture was allowed to stand at room temperature for 5 min and then Pseudomonas aeruginosa and Candida albicans [4]. sodium carbonate solution (75 g/L in water, 2 mL) was added. After 1 h of incubation, the absorbance was measured at 765 nm against Its ethyl acetate extracts exhibit cytotoxic activity [11]. In fact, blank using a Helios spectrophotometer (Unicam, Cambridge, UK). A Isoplumbagin isolated from the air-dried stem bark of Lawsonia standard calibration curve was plotted using gallic acid (0-300 mg/L). inermis exhibited cytotoxic activity (against melanoma and colon Results were expressed as g of gallic acid equivalents (GAE)/Kg of dry cancer cell lines as well as against several of the non-small cell lungs, mass. colon, CNS and renal cell lines) [12]. As we can see, this plant presents strong potential. That is why Condensed tannin content we seek to complement the knowledge of henna with chemical and Catechins and proanthocyanidins reactive with vanillin were biological studies. The aim of our work on Henna was to study to analyzed by the vanillin method [17]. One milliliter (1 mL) of each study its anatomical features, the chemical composition of its extracts extract solution was mixed in a test tube with 2 mL of vanillin (1% in 7 in the chemicals families such as polyphenols, tannins, antocyanins M H2SO4) in an ice bath. Then the mix was incubated at 25°C. After 15 and flavonoids. The activity of Henna extracts was also evaluated as an minutes, the solution absorbance was read at 500 nm. Concentrations antioxidant and anti-malarial. Finally, the toxicity against cancer cells were calculated as g catechin equivalents (CE)/Kg dry mass from a of human breast (MCF7) was also studied for these samples. calibration curve. Materials and Methods Total flavonoids determination Plant material The total flavonoids were estimated according to the Dowd method Lawsonia inermis L. collected in Morocco during 2009. Voucher as adapted by Arvouet-Grand [18]. A diluted methanolic solution (4 specimens of leaves identified by Professor C. Chatelain and deposited mL) of each extract was mixed with a solution (4 mL) of aluminium at the herbarium of the Faculty of Pharmacy in Toulouse. trichloride (AlCl3) in methanol (2%). The absorbance was read at 415 nm after 15 minutes against a blank sample consisting of a methanol (4 Preparing slides mL) and extract (4 mL) without AlCl3. Quercetin was used as reference Observations are based on microscopic studies of sectioned and compound to produce the standard curve, and the results were stained material of tissues. expressed as g of quercetin equivalents (QE)/Kg of dry mass. Transverse sections are prepared with a sliding microtome (MSE) Determination of total anthocyanin content and stained in alun carmine-green combination or Mirande reagent [13] during 2 to 3 minutes then washed with water.
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