Lawsonia Inermis – from Traditional Use to Scientific Assessment

The African Journal of Pharmaceutical Sciences and Pharmacy

AJPSP 2:1, 145-165, April 30, 2011

Lawsonia inermis – From Traditional Use to Scientific Assessment

Inder K. Makhija*1, D.R. Dhananjaya2, Vijay Kumar S1, Raviraj Devkar1, Devang Khamar1, Nishant Manglani3 and Sandeep Chandrakar3

1Department of Pharmacognosy, Manipal College of Pharmaceutical Sciences, Manipal University, Manipal-576 104, Karnataka 2Department of Pharmacology, Mallige College of Pharmacy, Banglore-560 090, Karnataka. 3Department of Pharmacognosy, JSS College of Pharmacy, Ooty-643 001, TamilNadu. *Corresponding author email: [email protected]

ABSTRACT

The world is endowed with a rich heritage of medicinal plants. The use of medicinal agents presumably predates the earliest recorded history. Medicinal plants are widely used by traditional practitioners for various ailments. Lawsonia inermis (Lythraceae) commonly known as ‘Henna’ is a well-known plant used in Indian medicine. Various parts of this plant have been used in traditional Indian medicine. The plant has a wide range of phytochemicals including lawsone, isoplumbagin lawsoniaside, lalioside, lawsoniaside B, syringinoside, daphneside, daphnorin, agrimonolide 6-O-β-D-glucopyranoside, (+)-syringaresinol O-β-D-glucopyranoside, (+)-pinoresinol di-O-β-D-glucopyranoside, syringaresinol di-O-β-D-glucopyranoside, isoscutellarin3β, hennadiol, (20S)-3β, 30-dihydroxylupane, lawnermis acid, 3-methyl-nonacosan-1-ol, laxanthones I, II, III and lacoumarin. Various in-vitro and in-vivo studies of L. inermis have reported the plant to have antibacterial, antifungal, antiparasitic, antiviral, anticancer, antidiabetic, tuberculostatic, anti- inflammatory, antifertility and wound healing properties. This review discusses the botany, traditional use, phytochemistry and pharmacological data of the plant.

KEYWORDS: Traditional uses, Phytochemical, Pharmacology, Lawsonia inermis

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INTRODUCTION Henna grows better in tropical savannah and tropical arid zones, in latitude between 15°

and 25° N and S, produces highest dye content in In the past, medicinal plants were temperature between 35-45°C. The optimal soil used intensively in folkloric medicine for temperature range for germination is 25-30°C. treatment of various disorders. Today, it is Henna leaves are very popular natural dye to estimated that about 80 % of people in colour hand, finger, nails and hair. The dye developing countries rely on traditional molecule, lawsone is the chief constituents of the medicines for their primary health care. plant; its highest concentration is detected in the Traditional medicines are becoming petioles (0.5-1.5 %). In folk medicines, henna has popular, due to high toxicity and adverse been used as astringent, anti-hemorrhagic, effects of orthodox medications. This has intestinal antineoplastic, cardio-inhibitory, led to the sudden increase in the number of hypotensive, sedative and also as therapeutic herbal industries in the drug market. Several against amoebiasis, headache, jaundice and plant species are used by various leprosy (10-11). indigenous systems such as Siddha,

Ayurveda, Unani and Allopathy for the treatment of different ailments (1-7). This MORPHOLOGY review emphasizes the traditional uses and biological properties of Lawsonia inermis. Lawsonia inermis is a glabrous branched shrub or small tree (2 to 6 m in height). Leaves Lawsonia inermis Linn (Lythraceae) are small, opposite, entire margin elliptical to is a perennial plant commonly known as broadly lanceolate, sub-sessile, about 1.5 to 5 cm Henna, having different vernacular names long, 0.5 to 2 cm wide, greenish brown to dull in India viz., Mehndi in Hindi, Mendika, green, petiole short and glabrous acute or obtuse Journal of Pharmaceutical Sciences of and Pharmacy Sciences Pharmaceutical Journal Rakigarbha in Sanskrit, Mailanchi in apex with tapering base. New branches are green Malayalam, Muruthani in Tamil, Benjati in in colour and quadrangular, turn red with age.

Oriya, Mayilanchi in Kannada and Mehedi Young barks are greyish brown, older plants have

African

in Bengali (8). It is native to North Africa spine-tipped branchlets (Fig. 1). Inflorescence has and South East Asia, and often cultivated as large pyramid shaped cyme. Flowers are small, an ornamental plant throughout India, numerous, aromatic, white or red coloured with AJPSP: Persia, and along the African coast of the four crumbled petals. Calyx has 0.2 cm tube and Mediterranean Sea (9). 0.3 cm spread lobes. The fruits are small, brown globose capsule, opening irregularly and split into four sections with a permanent style. Seeds have typical, pyramidal, hard and thick seed coat with brownish coloration (12-14).

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Figure 1: Morphology of Lawsonia inermis plant

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ETHNOPHARMACOLOGY

the Indian traditional folk medicines is well documented. Table 1 indicates the use of different AJPSP: Lawsonia inermis is a well known ethnomedicinal plant used cosmetically and parts of L. inermis in traditional system of medicinally for over 9,000 years. Its use in medicines.

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Table 1: Ethnomedicinal uses of different parts of L. inermis

Plant Parts Traditional Uses (as/in) Root Bitter, depurative, diuretic, emmenagogue, abortifacient, burning sensation, leprosy, skin diseases, amenorrhoea, dysmenorrhoea and premature greying of hair (15).

Leaves Bitter, astringent, acrid, diuretic, emetic, edema, expectorant, anodyne, anti- inflammatory, constipating, depurative, liver tonic, haematinic, styptic, febrifuge, trichogenous, wound, ulcers, strangury, cough, bronchitis, burning sensation, cephalalgia, hemicranias, lumbago, rheumatalgia, inflammations, diarrhoea, dysentery, leprosy, leucoderma, scabies, boils, hepatopathy, splenopathy, anemia, hemorrhages, hemoptysis, fever, ophthalmia, amenorrhoea, falling of hair, greyness of hair, jaundice (15-18).

Flowers Cardiotonic, refrigerant, soporific, febrifuge, tonic, cephalalgia, burning sensation, cardiopathy, amentia, insomnia, fever (15).

Seeds Antipyretic, intellect promoting, constipating, intermittent fevers, insanity, amentia, diarrhea, dysentery and gastropathy (15).

Journal of Pharmaceutical Sciences of and Pharmacy Sciences Pharmaceutical Journal PHYTOCHEMISTRY OF L. inermis are napthoquinone derivatives, phenolic

compounds, terpenoids, sterols, aliphatic African

INERMIS

derivatives, xanthones, coumarin, fatty acids, amino acids and other constituents. Phytochemicals reported in L. inermis L. are listed Much work is done in the field AJPSP: in (Table 2), with their structures in (Fig. 2). of phytochemical investigation of the plant. The chemical constituents isolated from L.

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Table 2: Phytochemicals found in various parts of L. inermis

Compounds Plant Parts References Napthoquinone derivatives (19) Lawsone (2-hydroxy 1,4-naphthoquinone) Leaves 1,3-dihydroxy naphthalene, 1,4-napthaquinone, 1,2-dihydroxy-4- Leaves (20) glucosylnaphthalene Isoplumbagin Stem bark (21) Phenolic conpounds Lawsoniaside (1,3,4-trihydroxynaphthalene 1,4-di-β-D-gluco-pyronoside), Bark, Leaves (22,23) Lalioside (2,3,4,6-tetrahydroxyacetoxy-2-β-D-glucopyranoside) Lawsoniaside B (3-(4-O-a-D-glucopyranosyl-3,5-dimethoxy) phenyl-2E- propenol), syringinoside, daphneside, daphnorin, agrimonolide 6-O-β-D- glucopyranoside, (+)-syringaresinol O-β-D-glucopyranoside, (+)- pinoresinol di-O-β-D-glucopyranoside, syringaresinol di-O-β-D- glucopyranoside, isoscutellarin Terpenoids 3β, 30-dihydroxylup-20(29)-ene (hennadiol), (20S)-3β, 30- Bark, Seeds (24, 25) dihydroxylupane, Lupeol, 30-nor-lupan-3β-ol-20-one, betulin, betulinic acid, lawnermis acid (3β-28β-hydroxy-urs-12,20-diene-28-oic acid) and its methyl ester Sterols Lawsaritol ( 24β-ethycholest-4-en-3β-ol) Roots, Leaves (26) Stigmasterol and β-sitosterol Aliphatic constituents 3-methyl-nonacosan-1-ol, n-tricontyl n-tridecanoate Stem bark (27, 28) Xanthones

Journal of Pharmaceutical Sciences of and Pharmacy Sciences Pharmaceutical Journal Laxanthone I (1,3 dihydroxy-6,7 dimethoxy xanthone), Laxanthone II (1- Whole plant (29,30) hydroxy-3,6 diacetoxy-7-methoxyxanthone), Laxanthone III ( 1-hydroxy- 6-acetoxy xanthone)

African Coumarins

Lacoumarin (5-allyoxy-7-hydroxycoumarin) Whole plant (31) Flavonoids Apigenin-7-glucoside, apigenin-4-glycoside, luteolin-7-glucoside, luteolin- Leaves (32)

AJPSP: 3-glucoside Essential oil -(Z)-2-hexenol, linalool, α ionone, β ionone, α-terpineol, terpinolene, δ-3- Leaves (33, 34) carene and γ-terpineol Other chemical constituents Hennotannic acid, glucose, gallic acid, amino acid Whole plant (35) Trace metal – Cu, Ni, Mo, V, Mn, Sr, Ba, Fe and Al Minerals – Na2O, CaO and K2O

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Figure 2: Shows biologically active constituents present in L. inermis

Lawsone Isoplumbagin Lawsoniaside

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Lalioside AJPSP:

Lawsoniaside B, R = α-D-glucopyranoside

Syringinoside, R = β-D-glucopyranosyl-(1 6)-β-D-glucopyranoside

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Daphneside

Daphnorin

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AJPSP:

Agrimonolide 6-O-β-D-glucopyranoside

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1 2 (+)-Syringaresinol O-β-D-glucopyranoside, R = OCH3, R = H

1 2 (+)-Pinoresinol di-O-β-D-glucopyranoside, R = H, R = β-D-glucopyranoside

1 2

Syringaresinol di-O-β-D-glucopyranoside, R = OCH3, R = β-D-glucopyranoside

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Isoscutellarin Hennadiol AJPSP:

Lupeol 30-nor-lupan-3β-ol-20-one

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Betulin Betulinic acid

Laxanthone - I Laxanthone - II Journal of Pharmaceutical Sciences of and Pharmacy Sciences Pharmaceutical Journal

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Laxanthone – III Lacoumarin AJPSP:

Apigenin-7-glucoside

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Luteolin Stigmasterol

β-sitosterol Linalool

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AJPSP:

α-terpineol α-ionone β-ionone Gallic acid

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PHARMACOLOGICAL found to be significantly elevated in liver. Within the extrahepatic organs examined (forestomach, ACTIVITY OF L. INERMIS kidney and lung) glutathione S-transferase and DT-diaphorase levels were increased in a dose Several researchers have reported independent manner (37). Another study was the different biological actions of L. inermis carried out to assess the effect of aqueous and

in various in-vitro and in-vivo test models. methanolic extracts of L. inermis extract on

Henna leaves, flower, seeds, stem bark, chromium (VI)–induced cellular and DNA roots have been found to exhibit toxicity. The extracts showed significant (P<0.05) antioxidant, antidiabetic, hepatoprotective, potential in scavenging free radicals of DPPH, hypoglycemic, antimicrobial, anticancer ABTS, Fe3+ and also in inhibition of lipid and wound healing properties. These are peroxidation. Extracts also showed significantly, described in greater details in the following properties of DNA and cyto-protection (38). sections. Different constituents isolated from the leaves of L. inermis were tested for their antioxidant ANTIOXIDANT ACTIVITY activity using ABTS. The IC50 value of henna constituents are p-coumaric acid (2.6 mM), Lawsone (2-hydroxy-1, 4 cosmosiin (2.9 mM) apiin (1.6 mM) respectively. napthoquinone) is the main ingredient of L. These fundings depicted that all isolated inermis. On oxidation of 100 µM compounds exhibited antioxidant activity phenanthridine by guinea pigs aldehyde comparable to that of ascorbic acid (2.5 mM) oxidase, superoxide anion and hydrogen (39). peroxide formation was found to be 6-10 %

and 85-90 % respectively. The mechanism ANTIBACTERIAL ACTIVITY of action was believed to be flavin

Journal of Pharmaceutical Sciences of and Pharmacy Sciences Pharmaceutical Journal semiquinone (FADH). The IC50 value of Forty-five species of 29 plant families lawsone was 9.3 ± 1.1 µm, which in excess used in the traditional medicine by Iranian people

of 15 fold of maximal plasma concentration African

showed antibacterial activities against bacterial of lawsone, indicating high degree of safety species, Bacillus cereus, Bacillus pumilus, margin (36). The modulator effect of 80 % Bordetella bronchiseptica, Escherichia coli, ethanol extract of leaves of L. inermis were

AJPSP: Kiebsiella pneumonia, Micrococcus luteus, studied on drug metabolizing phase-I and Pseudomonas aeruginosa, Pseudomonus phase-II enzymes, antioxidant enzymes, fluorescens, Serratia marcescens, Staphylococcus glutathione content, lactate dehydrogenase aureus and Staphylococcus epidermidis. Henna and lipid peroxidation in the liver of swiss showed strong activity against Bordetella albino mice. The hepatic glutathione S- bronchiseptica. These findings indicated that L. transferase and DT-diaphorase specific inermis can be used in the treatment of bacterial activities were elevated above basal level infections (40). Crude extracts of fresh dry leaves by L. inermis extract treatment. With and seeds were investigated for their antimicrobial reference to antioxidant enzymes the activity against three standard strains and the investigated doses were effective in eleven patients’ isolated strains. Henna dry leaves increasing the hepatic glutathione reductase demonstrated the best in-vitro antimicrobial (GR), superoxide dismutase (SOD) and activity and in particular against Shigella sonnei catalase activities significantly at both the (41). Ethanol extracts of 20 plants species used by

dose levels. Reduced glutathione (GSH) Yemeni traditional healers to treat infectious measured as non-protein sulphydryl was

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diseases were screened for their schenckii. Ethanol, methanol and aqueous extract antibacterial activity. The ethyl acetate of leaves of L. inermis are involved in defensive extract of L. inermis was found to be the mechanism against spore germination of most active against all the bacteria in the Drechslera oryzae (45). During screening of barks test system (42). Genotoxic studies on main of 30 plant species for activity against constituent of henna suggested that it was a Microsporum gypseum and Trichophyton

weak bacterial mutagen for Salmonella mentagrophytes, only L. inermis extract exhibited

typhimurium strain TA98 and was more absolute toxicity. The extract showed broad mutagenic for strain TA2637. It suggested fungitoxic spectrum when tested against 13 other that hydroxyl napthaquinone have no dermatophytes. Further the fungitoxicity of the genotoxic risk to the consumer (43). extract remained unaltered at high temperature on Primary invaders of burn wounds viz autoclaving and after long storage (46). Aqueous Staphylococcus aureus, Pseudomonas extract of leaves of L. inermis was tested for the aeruginosa, Candida albicans, Fusarium antifungal potential against eight important oxysporum and Aspergillus niger were species of Aspergillus which were isolated from treated with aqueous and chloroform extract sorghum, maize and paddy seed samples. A. obtained from leaves of L. inermis, using flavus recorded high susceptibility and hence in-vitro agar incorporation and well solvent extracts viz., petroleum ether, benzene, diffusion methods. Extract inhibit growth chloroform, methanol and ethanol extract of the pattern of all microbes except C. albicans. plant showed significant antifungal activity (47). Overall, study suggested that henna may be These finding suggested that henna extract could effective in the management of wound be used as alternative source of antifungal agents infections (35). The antibacterial activity of for protection of plants or crops against fungal methanolic extract of L. inermis was infection.

Journal of Pharmaceutical Sciences of and Pharmacy Sciences Pharmaceutical Journal investigated by agar well diffusion method using S. aureus (MTCC 087), E. coli HEPATOPROTECTIVE ACTIVITY

(MTCC 729), K. pneumonia (MTCC 432), African

P. aeruginosa (MTCC 1688) and P. Alcoholic extract of the bark of L. inermis

mirablis (MTCC 425). According to results showed hepatoprotective effect against CCl4. The in the lowest tested concentration of 62.5 extract causes elevation in serum marker enzymes

AJPSP: μg/ml and 125 μg/ml, 7.2 % of the plant (GOT and GPT), serum bilirubin, liver lipid extract were active, 5 % active in the peroxidation and reduction in total serum protein, concentration of 250 μg/ml, 75.7 % active liver glutathione, glutathione peroxidase, in the concentration of 500 μg/ml and 92.8 glutathione-s-transferase, glycogen, superoxide % active at the concentration of 1000 μg/ml dismutase and catalase activity (16,48,49). The in a dose dependent manner (44). hepatoprotective activity of the ethanolic extract of the dried leaves of L. inermis and its fractions ANTIFUNGAL ACTIVITY (petroleum ether, ethyl acetate, butanol and

butanone fractions) was evaluated against CCl4 Sensitivity toward henna was strong induced hepatotoxicity in mice. The ethanolic in Trichophyton mentagrophytes, T. extract and its fractions significantly reduced the rubrum, T. tonsurans, T. violaceum, T. total bilirubin content and SGOT, SGPT and SAL verrocosum, T. schoenleinii, activities, and reduced liver weight compared to Epidermophyton floccosum, Microsporum LIV-52 used as control (50, 51).

ferrugineum, M. canis, sporotrichum

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high skin breaking strength, a significant increase ANTIDIABETIC ACTIVITY in the granulation tissue weight and hydroxyproline content. Histological studies of Ethanol (70 %) extract of L. inermis the tissue showed increased well organized bands showed significant hypoglycemic and of collagen, more fibroblasts and few hypolipidemic activities in alloxan induced inflammatory cells when compared with the

diabetic mice after oral administration. The controls which showed inflammatory cells, scanty

feeding of 0.8 g/kg of L. inermis extract collagen fibres and fibroblasts. These findings decreased the concentration of glucose, suggest the use of L. inermis in the management cholesterol and triglycerides to normal (52). of wound healing (54). Chloroform and aqueous extracts of leaves of the plant were capable of PROTEIN GLYCATION INHIBITORY inhibiting the growth of microorganisms that are ACTIVITY involved in causing burn wound infections (55, 35). Ethanol extract of the plant tissues was evaluated in-vitro for protein glycation ANTI-INFLAMMATORY ACTIVITY inhibitory activity using the model system of bovine serum albumin and glucose. The Isoplumbagin and lawsaritol, isolated extract and its components showed from stem bark and root of L. inermis was significant effect on protein damage screened for anti-inflammatory activity against induced by a free radical generator during carrageenan induced paw edema in rats. The in-vitro assay system. It was found that the compounds phenylbutazone, isoplumbagin and alcoholic extract, lawsone and gallic acid lawsaritol at the oral dose of 100 mg/kg exhibited showed significant inhibition of Advanced 61, 60 and 40 % inhibition respectively. The

Journal of Pharmaceutical Sciences of and Pharmacy Sciences Pharmaceutical Journal Glycated End Products (AGEs) formation results showed that isoplumbagin, which and exhibit 77.95 %, 79.10 % and 66.98 % exhibited significant activity, was compared to

inhibition at a concentration of 1500 that of phenylbutazone (56). Butanol and African

μg/mL, 1000 μg/mL and 1000 μgM/ml chloroform fractions showed potent, anti- respectively. L. inermis constituents were inflammatory, analgesic, and antipyretic effects found to be glycation inhibitors with IC50 than aqueous fraction of crude ethanol extract of

AJPSP: 82.06±0.13 μg/mL, 67.42±1.46 μM and L. inermis in a dose dependent manner (57). 401.7±6.23 μM respectively (53). TUBERCULOSTATIC ACTIVITY WOUND HEALING ACTIVITY The tuberculostatic activity of henna was Ethanol extract of the plant (200 tested in-vitro and in-vivo using Lowenstein mg/kg/) was used to evaluate the wound Jensen medium; the growth of Tubercle bacilli healing activity on rats using excision, from sputum and of Mycobacterium tuberculosis incision and dead space wound models. H37Rv was inhibited by 6 μg/ml of the herb. In- Topical application was made in the case of vivo studies on guinea pigs and mice showed that excision wound model. Whereas, oral the plant at a dose of 5 mg/kg body weight led to treatment was done with incision and dead a significant resolution of experimental space wound model. Extract of L. inermis tuberculosis following infection with M. showed high rate of wound contraction, a tuberculosis H37Rv (58).

decrease in the period of epithelialization,

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(CP) alone and in combination with L. inermis ANTIVIRAL ACTIVITY was studied in B16F10 melanoma tumour bearing mice. The Inhibition rate was 25.9% in the CP The ethanol soluble fraction of L. treated group but these increased to 35.14% with inermis fruits displayed highly potent L. inermis. The life span time and volume of activity against Sembiki forest virus (SFV) tumour doubling time were also increased. Results

in Swiss mice and chick embryo models from two models showed that L. inermis extract

exhibiting 100 to 65 % activities after 10 to have protective potential against skin tumour (62). 25 days of virus challenge (59).

ANTICANCER ACTIVITY MOLLUSCICIDAL ACTIVITY

The anticarcinogenic activity of The molluscicidal activity of leaf, bark and chloroform extract L. inermis leaves was seed of henna against Lymnaea acuminata and carried using microculture tetrazolium salt Indoplanorbis exustus were studied. Seed powder assay on the human breast (MCF-7), colon was more toxic than leaf and bark against L. (Caco-2), liver (HepG2) carcinoma cell exustus. Binary combinations of henna seed with lines and normal human liver cell lines Cedrus deodara Roxh and Azadirachta indica A (Chang Liver). The preliminary results Juss oil, or powdered Allium sativum, or Zingiber showed that the henna extract displayed the officinale rhizome oleoresin was more toxic to cytotoxic effects against HepG2 and MCF-7 snails L. acuminata and I. exustus than their single and IC-value of 0.3 and 24.85 µg/ml treatment. The highest increase in the toxicity was respectively (60). Isoplumbagin at a observed when henna seeds powder and C. concentration of 10.5–10.8 M, the deodara oil (1:1) were tested against both the

Journal of Pharmaceutical Sciences of and Pharmacy Sciences Pharmaceutical Journal compound typically produced LC50 – level snails. The combination with neem oil was also responses against a majority of the more toxic than their individual components and

melanoma and colon cancer cell lines as other combinations (63). African

well as against several of the non-small cell

lungs, colon, CNS, and renal cell lines. ANTITRYPANOSOMAL ACTIVITY Isoplumbagin showed an interesting profile

AJPSP: of cytotoxic activity (61). Crude methanolic extract of leaf of L. inermis showed in-vitro activity against The antitumour activity of L. Trypanosoma brucei at a concentration of 8.3 inermis leaf extract was studied on 7, 12- mg/ml of blood in mice but not in-vivo. The dimethylbenzanthracene (DMBA) induced treatment tended to ameliorate the disease 2-stage skin carcinogenesis and B16F10 condition, but did not affect the level of melanoma tumour model using swiss albino parasitaemia and packed cell volume (64). mice. Topical application of L. inermis leaf extract at a dose level of 1000 mg/kg body ANTIDERMATOPHYTIC ACTIVITY weight was found to be effective in reducing the number of the papillomas. The The antidermatophytic activity of ethanol, tumor yield was significantly decreased to ethyl acetate and hexane extracts of L. inermis 1.6 as compared with the DMBA treated were tested on 5 strains each of Tinea rubrum and control group decrease of 3.5. In other Tinea mentagrophytes. All these extracts showed

experiment the effect of cyclophosphamide

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significant antidermatophytic properties in- assay (LTA)-guided fractionation of the total vitro (65). methanolic extract of henna leaves (39). ANTIPARASITIC ACTIVITY Naphthoquinone fraction obtained from leaves of L. inermis showed significant immunomodulatory During an ethnopharmacological effect (68). survey of antiparasitic medicinal plants

used in Ivory Coast, 17 plants were

identified and collected. Polar, non-polar NOOTROPIC ACTIVITY and alkaloidal extracts of various parts of these species were evaluated in-vitro in an The effect of acetone soluble fraction of antiparasitic drug screening. Antimalarial, petroleum ether extract of L. inermis leaves was leishmanicidal, trypanocidal, investigated on memory, anxiety and behaviour antihelminthiasis and antiscabies activities mediated via monoamine neurotransmitters using were determined. Among the selected elevated plus maze and passive shock avoidance plants, leaves of L. inermis showed paradigms. The extract exhibited prominent potential trypanocidal activities (66). nootropic activity, potentiated clonidine induced hypothermia and decreased lithium induced head ANTIFERTILITY ACTIVITY twitches. However, haloperidol induced catalepsy was not modified (69). Ethanol extract prepared from the powdered seeds of L. inermis failed to show CONCLUSION significant antifertility activity. However in subsequent studies it was observed that the powdered leaves of henna, when Lawsonia inermis is not only a colouring agent, but it also possesses various biological

Journal of Pharmaceutical Sciences of and Pharmacy Sciences Pharmaceutical Journal administered as suspension or incorporated into the diet inhibited the fertility of rats. activities such as antioxidant, antimicrobial, The fertility induced appeared was found to antidiabetic, anticancer, anti-inflammatory,

antiparasitic, antidermatophytic properties, African

be permanent (67). anticancer, antiviral, wound healing,

immunomodulatory, hepatoprotective, IMMUNOMODULATORY ACTIVITY tuberculostatic, antifertility, and protein glycation

AJPSP: Methanol extract of henna leaves at 1 inhibitor properties. Scientific research on L. mg/ml concentration had displayed inermis suggested huge biological properties of immune-stimulant action as indicated by the extracts might provide detailed evidence for promotion of T-lymphocyte proliferative the use of this plant in different medicines. responses. Seven compounds were isolated Furthermore clinical trials on the use of this plant adopting the lymphocyte transformation need to be conducted.

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REFERENCES

1. Farnsworth NR, Akerele O, Bingel AS, Soejarto DD, Guo Z. Medicinal plants in therapy. Bulletin of the World Health Organization 1985:63(6):965-981.

2. Verpoorte R. Pharmacognosy in the new millennium: lead finding and biotechnology.

J. Pharm. Pharmacol. 2000:52(3):253-262.

3. Turner DM. Natural product source material use in the pharmaceutical industry: the Glaxo experience. J. Ethnopharmacol. 1996:51(1-3):39-44.

4. Newman DJ, Cragg GM, Snader KM. The influence of natural products upon drug discovery. Nat. Prod. Rep. 2000:17(3):215-234.

5. Clark AM. Natural products as a resource for new drugs. Pharm. Res. 1996: 13(8):1133-1144.

6. Cragg GM, Newman DJ, Snader KM. Natural products in drug discovery and development. J. Nat. Prod. 1997:60(1):52-60.

7. Cowan MM. Plant Products as antimicrobial agents. Clin. Microbiol. Rev. 2009:12(4):564-582. 8. Kirtikar KR, Basu BD. Indian Medicinal Plants, Vol. 3. International book distributors,

Journal of Pharmaceutical Sciences of and Pharmacy Sciences Pharmaceutical Journal New Delhi, India, 1956.

9. Malekzadeh F. Antimicrobial Activity of Lawsonia inermis L. American Society for

African

Microbiology 1968:16(4):663-634.

10. Rao SS, Regar PL, Singh YV. Agrotechniques for henna (Lawsonia inermis L.)

AJPSP: cultivation, improvement and trade. Central Arid Zone Research Institute, Pali-Marwar, 2005, pp: 25-27.

11. Simon JE, Chadwick AF, Craker LE. In: Herbs an indexed bibliography: The scientific literature on selected herbs aromatic and medicinal plants of the temperate zone. Hamden: Archon Books; 1984. P.1971-1980.

12. Sastri BN. The Wealth of India: Raw Materials, CSIR, New Delhi: 1962. P. 47-50.

13. Chauhan MG, Pillai APG. Microscopic profile of powdered drug used in Indian system of medicine, Jamnagar, Gujarat: 2007. P. 84-85.

14. Vasudevan TN, Laddha KS. Herbal drug microscopy, Yucca publishing house, Dombivli: 2003. P. 68-69.

AJPSP 2011; 2:1 Makhija et al Page 160

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15. Warrier PK. Indian medicinal plants a compendium of 500 species, Vol. 3. Chennai: Orient longman private limited; 2004. P. 303-304.

16. Ahmed S, Rahman A, Alam A, Saleem M, Athar M, Sultana S. Evaluation of the efficacy of Lawsonia alba in the alleviation of carbon tetrachloride induced oxidative stress. J. Ethnopharmacol. 2000:69(2):157-164.

17. Bich DH, Chung DQ, Chuong BX, Dong NT, Dam DT, Hien PV, Lo VN, Mai PD, Man PK, Nhu DT, Tap N, Toan T. The Medicinal Plants and Animals in Vietnam, Vol. 2. , Hanoi, Vietnam: Science and Technology Publishing House; 2004.

18. Reddy KR. Folk Medicine from Chittoor District, Andhra Pradesh, India, Used in the Treatment of Jaundice. International Journal of Crude Drug Research 1988:26(3):137- 140.

19. Dixit SN, Srivastava HS, Tripathi RD. Lawsone, the antifungal antibiotic from the leaves of Lawsonia inermis and some aspects of its mode of action. Indian Phytopathological 1980:31:131-133.

20. Afzal M, Al-oriquat G, Al-Hassan JM, Muhammad N. Isolation of 1,2-Dihydroxy-4- glucosyloxynaphthalene from Lawsonia inermis. Heterocycles 1984:22(4):813.

21. Gupta S, Ali M, Alam MS. A napthaquinone from lawsonia inermis stem bark. Phytochemistry 1993:33(3):723-724.

Journal of Pharmaceutical Sciences of and Pharmacy Sciences Pharmaceutical Journal 22. Takeda Y, Fatope MO. New phenolic glucosides from Lawsonia inermis. J. Nat. Prod.

1988:51(4):725-729. African

23. Cuong NX, Nhiem NX, Thao NP, Nama NH, Dat NT, Anh HLT, Huong LM, Kiem PV, Minh CV, Wonc J, Chung W, Kim YH. Inhibitors of osteoclastogenesis from

AJPSP: Lawsonia inermis leaves. Bioorg. Med. Chem. Lett. 2010:20(16):4782-4784.

24. Chakrabartty T, Poddar G, St-Pyrek J. Isolation of dihydroxy lupene and dihydroxy lupane from the bark of Lawsonia inermis. Phytochemistry 1982:21(7):1814-1816.

25. Handa G, Kapil A, Sharma S, Jagdev S. Lawnermis acid a new anticomplementary triterpenoids from Lawsonia inermis seeds. Indian J. Chem. 1997:36(3):252-256.

26. Gupta S, M Ali M, Alam MS, Niwa M, Sakai T. 24β-ethylcholest-4-en-3-β-ol from the Roots of Lawsonia inermis. Phytochemistry 1992:31(7):2558-2560.

27. Gupta S, Ali M, Alam MS, Sakai T, Niwa M. A new aliphatic hydrocarbon from Lawsonia inermis bark. Indian J. Chem. 1992:31:705-707.

AJPSP 2011; 2:1 Makhija et al Page 161

African Journal of Pharmaceutical Sciences and Pharmacy 2011;2:1

28. Chakrabartty D, Poddar G, Deshmukh SK. Triterpenoids and other constituents of Lawsonia inermis. Indian J. Chem. 1977:15:96-97.

29. Bhardwaj DK, Sheshadri TR, Singh R. Xanthones from Lawsonia inermis. Phytochemistry 1977:16(10):1616-1617.

30. Bhardwaj DK, Jain RK, Jain BC, Mehta CK. 1-hydroxy-3, 7- dimethoxy-6-acetoxy

xanthone a new xanthone from Lawsonia inermis. Phytochemistry 1978:17(8):1440- 1441.

31. Bhardwaj DK, Murari R, Sheshadri TR, Singh R. Lacoumarin from Lawsonia inermis. Phytochemistry 1976:15(11):1789.

32. Afzal M, Al-oriquat G, Al-Hassan JM, Muhammad N. Flavone glycosides from Lawsonia inermis. Heterocycles 1980:14:1973-1976.

33. Wong KC, Teng YE, Volatile Components of Lawsonia inermis L. Flowers. J. Essent. Oil Res. 1995:7(4):425-428.

34. Reichling VJ, Harkenthal M, Brandt H, Bayerl C. Temporare Henna Taottoos. Deutsche Apotheker Zeitung 1999:(139)33:35-41.

35. Muhammad HS, Muhammad S. The use of Lawsonia inermis linn. (Henna) in the management of burn wound infections. Afr. J. Biotechnol. 2005:4(9):934-937.

Journal of Pharmaceutical Sciences of and Pharmacy Sciences Pharmaceutical Journal 36. Omar MA. Effects of 2-hydroxy-1, 4-napthoquinone, a natural dye of henna, on

aldehyde oxidase activity in guinea pigs. Journal of Medical Science 2005:5(3):163- African

168.

37. Dasgupta T, Rao AR, Yadava PK. Modulatory effect of Henna leaf (Lawsonia inermis)

AJPSP: on drug metabolising phase I and phase II enzymes, antioxidant enzymes, lipid peroxidation and chemically induced skin and forestomach papillomagenesis in mice. Mol. Cell. Biochem. 2003:245(1-2):11-22.

38. Guha G, Rajkumar V, Kumar A, Mathew L. Antioxidant activity of Lawsonia inermis extracts inhibits chromium (VI)-induced cellular and DNA toxicity. Evid. Base. Compl. Alternative Med. 2009:6(4):1-10.

39. Mikhaeil BR, Badria FA, Maatooq GT, Amer MM. Antioxidant and immunomodulatory constituents of henna leaves. Zeitschrift für Naturforschung C 2004:59:468-476.

40. Bonjar S. Evaluation of antibacterial properties of some medicinal plants used in Iran. J. Ethnopharmacol. 2004:94(2-3):301-305.

AJPSP 2011; 2:1 Makhija et al Page 162

African Journal of Pharmaceutical Sciences and Pharmacy 2011;2:1

41. Habbal OA, Ai-Jabri AA, El-Hag AH, Al-Mahrooqi ZH, Al-Hashmi NA. In-vitro antimicrobial activity of Lawsonia inermis Linn (henna) - A pilot study on the Omani henna. Saudi Med. J. 2005:26(1):69-72.

42. Ali NAA, Julich WD, Kusnick C, Lindequist U. Screening of Yemeni medicinal plants for antibacterial and cytotoxic activities. J. Ethnopharmacol. 2001:74(2):173-179.

43. Kirkland D, Marzin D. An assessment of the genotoxicity of 2-hydroxy-1, 4- naphthoquinone, the natural dye ingredient of Henna. Mutation Research 2003:537(2):183-199.

44. Arun P, Purushotham KG, Jayarani J, Kumari V. In vitro Antibacterial activity and Flavonoid contents of Lawsonia inermis (Henna). International Journal of PharmTech Research, 2010:2(2):1178-1181.

45. Natarajan MR, Lalitha DK. Leaf extracts of Lawsonia inermis as antifungal agent. Current Science 1987:56(19):1021-1022.

46. Singh VK, Pandey DK. Fungitoxic studies on bark extract of Lawsonia inermis against ringworm fungi. Hindustan Antibiotics Bulletin 1989:31(1-2):32-35.

47. Raveesha KA, Satish S, Mohana DC, Raghavendra MP. Antifungal activity of some plant extracts against important seed borne pathogens of Aspergillus sp. Journal of Agricultural Technology 2007:3(1):109-119.

Journal of Pharmaceutical Sciences of and Pharmacy Sciences Pharmaceutical Journal 48. Anand KK, Singh B, Chand D, Chandan, BK. An evaluation of Lawsonia alba extract

as hepatoprotective agent. Planta Medica 1992:58(1):22-25. African

49. Bhandarkar M, Khan A. Protective effect of Lawsonia alba against carbontetrachloride induced hepatic damage in albino rats. Indian J. Exp. Biol. 2003:41(1): 85-87.

AJPSP: 50. Hemalatha K, Natraj HN, Kiran AS. Hepatoprotective activity of leaves of Lawsonia alba. Indian J. Nat. Prod. 2004:20(4):14-17.

51. Latha PG, Suja SR, Shyamal S, Rajasekharan S. Some hepatoprotective garden plants. Natural Product Radiance 2005:4(4):278-279.

52. Syamsudin I, Winarno H. The effects of Inai (Lawsonia inermis) leave extract on blood sugar level: An Experimental Study. Res. J. Pharmacol. 2008:2(2):20-23.

53. Sultana N, Choudhary MI, Khan AJ. Protein glycation inhibitory activities of Lawsonia inermis and its active principles. J. Enzym. Inhib. Med. Chem. 2009:24(1):257-261.

54. Nayak BS, Isitor G, Davis EM, Pillai GK. The evidence based wound healing activity

of Lawsonia inermis Linn. Phytotherapy Research 2007:21(9):827-831.

AJPSP 2011; 2:1 Makhija et al Page 163

African Journal of Pharmaceutical Sciences and Pharmacy 2011;2:1

55. Hamdi YP, Benazzouz M, Belkhiri H, Chari Z, Serakta M, Bensgni L. Healing effect of Lawsonia inermis L. (henna) as exemplified by the third degree burns. Revue de Medecines et Pharmacopees Africaines 1997:11-12:151-156.

56. Gupta S, Ali M, Pillai KK, Alam MS. Evaluation of anti-inflammatory activity of some

constituents of Lawsonia inermis. Fitoterapia 1993:64(4):365-366.

57. Ali BH, Bashir AK, Tanira MOM. Antiinflammatory, antipyretic and analgesic effects of Lawsonia inermis L. (henna) in rats. International Journal of Experimental and Clinical Pharmacology 1995:51(6):356-363.

58. Sharma VK. Tuberculostatic activity of henna Lawsonia inermis Linn. Tubercle 1990:71(4): 293-296.

59. Khan MMAA, Jain DC, Bhakuni RS, Zaim M, Thakur RS. Occurrence of some antiviral sterols in Artemisia annua. Plant Sci. 1991:75(2):161-165.

60. Endrini S, Rahmat A, Ismail P, Taufiq-Yap YH. Comparing of the cytotoxicity properties and mechanism of Lawsonia inermis and Strobilanthes crispus extract against several cancer cell lines. Journal of Medical Sciences 2007:7(7):1098-1102.

61. Ali M, Grever MR. A cytotoxic napthoquinone from Lawsonia inermis. Fitoterapia 1998:69(2):181-183.

Journal of Pharmaceutical Sciences of and Pharmacy Sciences Pharmaceutical Journal 62. Wasim R, Agrawal RC, Ovais M. Chemopreventive action of Lawsonia inermis Leaf

Extract on DMBA-induced skin papilloma and B16F10 Melanoma Tumour. African

Pharmacologyonline 2009:2:1243-1249.

63. Singh A, Singh DK. Molluscicidal activity of Lawsonia inermis and its binary and

AJPSP: tertiary combinations with other plant derived molluscicides. Indian J. Exp. Biol. 2001:39(3):263-268.

64. Wurochekke AU, Chechet G, Nok AJ. In-vitro and In-vivo antitrypanosomal brucei infection in mice. Journal of Medical Science 2004:4(3):236-239.

65. Natarajan V, Mahendraraja S, Menon T. Antidermatophytic activities of Lawsonia alba. Biomedicine 2000:20(4):243-245.

66. Okpekon T, Yolou S, Gleye C, Roblot F, Loiseau P, Bories C, Grellier P, Frappier F, Laurens A, Hocquemiller R. Antiparasitic activities of medicinal plants used in Ivory Coast. J. Ethanopharmaco. 2004:90(1):91-97.

67. Munshi SR, Shetye TA, Nair RK. Antifertility activity of three indigenous plant

preparations. Planta Medica 1977:31(1):73-75.

AJPSP 2011; 2:1 Makhija et al Page 164

African Journal of Pharmaceutical Sciences and Pharmacy 2011;2:1

68. Dikshit V, Dikshit J, Saraf M, Thakur V, Sainis K. Immunomodulatory activity of naphthoquinone fraction of Lawsonia inermis Linn. Phytomedicine 2000:7:102-103.

69. Iyer MR, Pal SC, Kasture VS, Kasture SB. Effect of Lawsonia inermis on memory and

behaviour mediated via Monoamine neurotransmitters. Indian J. Pharmacol.

1998:30(3):181-185.

Journal of Pharmaceutical Sciences of and Pharmacy Sciences Pharmaceutical Journal

African

AJPSP:

AJPSP 2011; 2:1 Makhija et al Page 165