590 Chiang Mai J. Sci. 2014; 41(3)

Chiang Mai J. Sci. 2014; 41(3) : 590-605http://epg.science.cmu.ac.th/ejournal/Contributed Paper

Wedelia trilobata L.: A Phytochemical andPharmacological ReviewNeelam Balekar [a,b], Titpawan Nakpheng [a] and Teerapol Srichana*[a,b][a] Drug Delivery System Excellence Center, Faculty of Pharmaceutical Sciences, Prince of Songkla University,

Hat-Yai, Songkhla 90112, Thailand.[b] Department of the Pharmaceutical Technology, Faculty of Pharmaceutical Sciences, Prince of Songkla

University, Hat-Yai, Songkhla 90112, Thailand.*Author for correspondence; e-mail: teerapol.s@psu.ac.th

Received: 8 November 2012Accepted: 11 July 2013

ABSTRACTStudies on the traditional use of medicines are recognized as a way to learn

about potential future medicines. Wedelia is an extensive genus of the family Asteraceae,comprising about 60 different species. Wedelia trilobata Linn. has long been used astraditional herbal medicine in South America, China, Japan, India and for the treatmentof a variety of ailments. The aim of this review was to collect all available scientificliterature published and combine it into this review. The present review comprises theethnopharmacological, phytochemical and therapeutic potential of W. trilobata. Anexhaustive survey of literature revealed that tannin, saponins, flavonoids, phenol,terpenoids constitute major classes of phytoconstituents of this plant. Pharmacologicalreports revealed that this plant has antioxidant, analgesic, anti-inflammatory,antimicrobial, wound healing, larvicidal, trypanocidal, uterine contraction, antitumor,hepatoprotective, and in the treatment of diabetes, menstrual pain and reproductiveproblems in women. W. trilobata seems to hold great potential for in-depth investigationfor various biological activities, especially their effects on inflammation, bacterialinfections, and reproductive system. Through this review, the authors hope to attractthe attention of natural product researchers throughout the world to focus on theunexplored potential of W. trilobata, and it may be useful in developing new formulationswith more therapeutic value.

Keywords: Wedelia trilobata (L.), ethnopharmacological use, phytochemistry,pharmacological activity

1. INTRODUCTIONThe manufacture and clinical evaluation

of herbal remedies and/or theirconstituents have made it possible totransform traditional medicine into amodern industry capable of making a

significant contribution to the delivery ofhealthcare [1]. Many medicinal plantscontain large amounts of antimicrobial andantioxidant compounds such as phenolsand flavonoids. Phenolic compounds are a

Chiang Mai J. Sci. 2014; 41(3) 591

group of plant metabolites that havenumerous beneficial activities such asanti-inflammatory, anti- bacterial, anti-mutagenic, anti-viral and antioxidantproperties [2]. This revival of interest inplant derived drugs is mainly due to thecurrent widespread belief that “greenmedicine” is safe, and clinically effective,better tolerated by patients, less expensiveand globally competitive [3,4]. Use ofmedicinal plants is becoming popular inUnited States and Europe [5] and in mostof the developing world, plants or herbalproducts have forever played importantroles in the treatment of wounds, intestinalproblems, coughs and sneezes, generaltorpor etc [6].

Traditional knowledge about medicinalplants has without doubt served as anexample for developing new drugs. Overthe past decades, it has been shown thatWedelia trilobata L. contains high amountof diterpene, eudesmanolide lactones andluteolin with a variety of biological activities.This review assesses the scientific informationon W. trilobata and focuses on the evidenceavailable on its therapeutic benefits.

Wedelia, known officially by the scientificname, Sphagneticola trilobata (L.) Pruski, but stillcommonly referred to by its former name,W. trilobata (L.) Hitchc is a member of thefamily Asteraceae (formerly Compositae), thesunflower or daisy family. W. trilobata is a soilcreeper and forms a thick carpet. The genusWedelia, named in honor of Georg WolfgangWedel (1645-1721), Professor of Botany atJena, Germany, has about 70 species oftropical and subtropical regions. W. trilobatais a very attractive plant because of its nearlyconstant and prolific blooming [7].

2. DESCRIPTION2.1 Habitat

A weed of urban bushland, closed

forests, forest margins, open woodlands,waterways, lake margins, wetlands,roadsides, disturbed sites, waste areas,vacant lots, and coastal sand dunes intropical and sub-tropical regions. It mayalso encroach into lawns, footpaths andparks from nearby gardens [7, 8].

2.2 Geographical DistributionNative to Mexico, Central America

(i.e. Belize, Costa Rica, Guatemala,Honduras, Nicaragua and Panama), the andthroughout the Caribbean, where it is notedas a weed in Trinidad, Puerto Rico, theDominican Republic, Jamaica, Panama, andtropical South America (i.e. French Guiana,Guyana, Surinam, Venezuela, Brazil, Bolivia,Colombia, Ecuador and Peru). Naturalized inSouth Africa, Florida, Louisiana, Hawaii, PuertoRico, and the Virgin Islands. Escaped in manytropical regions of the world, includingAustralia (South-eastern Queensland andnorth-eastern New South Wales), the PacificIslands (i.e. American Samoa, the CookIslands, Fiji, French Polynesia, Guam,Kiribati, the Marshall Islands, Nauru,Niue, New Caledonia, Palau, WesternSamoa, Tonga and Hawaii), Malaysia,Indonesia, Thailand, India, Papua NewGuinea [9, 10].

2.3 Scientific ClassificationKingdom: Plantae – Plants; Subkingdom:

Tracheobionta – Vascular plants; Super-division: Spermatophyta – Seed plants;Division: Magnoliophyta – Floweringplants; Class: Magnoliopsida – Dicotyledons;Subclass: Asteridae; Order: Asterales;Family: Asteraceae – Aster family; Genus:Sphagneticola O. Hoffmann – creeping-oxeye; Species: Sphagneticola trilobata (L.)Pruski – Bay Biscayne creeping-oxeye [9].

592 Chiang Mai J. Sci. 2014; 41(3)

2.4 SynonymsComplaya trilobata (L.) Strother, Silphium

trilobatum L., Thelechitonia trilobata (L.) H.Rob.& Cuatrec., Wedelia carnosa Rich., Wedeliapaludosa DC., Wedelia trilobata (L.) A.S. Hitchc.,Wedelia triloba (Rich.) Bello [9].

2.5 Common NamesAtiat (Puluwat), Bay Biscayne creeping

oxeye, creeping daisy, creeping wedelia,rabbit’s paw, Singapore daisy, gold cup, yellowdots, trailing daisy, water zinnia and wildmarigold (English), hansenfuss (Germany),America hama-guruma (Japan), di jin hua(China), kra dum tong (Thailand), wedeliakuning (Malaysia), Singapore-madeliefie(Africa), ampelkrage (Sweden), ut telia(Marshall Islands), and arnica-do-mato,pseudo-arnica, vedelia (Brazil) [10].

2.6 Botanical DescriptionIt is a long lived (perennial) herb with

a creeping or climbing habit. This mat-forming herb often creates a dense groundcover (usually 15-30 cm tall but occasionallyupto 70 cm tall) that crowd out the growthof other species. It may also climb a shortdistance up trees or over other vegetation.The stems are rounded, green or reddishin color, and may be coarsely hairy. Theygrow up to 2 m long and regularly developroots (adventitious roots) at their nodes.Short, semi-upright (ascending), floweringbranches are produced of these creepingstems. The leaves are attractive, bright shinygreen, somewhat fleshy oppositely arranged and simple, the blade obovate to elliptic orovate and are stalkless (sessile) or borneon short stalks (petioles). These leaves 2-9cm long and 2-5 cm wide, acute at the apexand winged and sessile at the base usually havethree lobes (hence the name trilobata) andirregularly toothed (serrated) margins. They areglossy in appearance, mostly hairless

(glabrous), and slightly fleshy (succulent)in nature. The single attractive bright-yellow flower heads are daisy-like inappearance and are borne on the end ofterminal and axillary stalks (peduncles) 2to 9 cm long, with 2 to 4 series of bractsforming the involucre at the base of theflower. Each flower-head has 8-13 yellowish‘petals’ (ray florets) that are 6-15 mm longwith 1- to 3 finely toothed tips and arepistillate. In the centre of these flower-headsthere are numerous tiny yellow tubular discflorets 4-5 mm long, and mixed with chaffybracts. The ray and disc florets are bothyellow. The base of each flower-head (capitulum) is enclosed in a row (involucre)of narrow (lanceolate) green bracts (about 1cm long). Flowering occurs throughout theyear, but is most common from spring toautumn. The fruit is a 2 to 4-angled achene,with short, narrow pappus scales on the top[11]. The ‘seeds’ (i.e. achenes), when present,are 4-5 mm long and topped with a crownof short fringed scales. They are elongated inshape, brown in colour and have a roughsurface texture. However, very few seedsreach maturity in cultivated or naturalisedplants in Australia [12].

Figure 1. Photograph of Wedelia trilobataLinn.

Chiang Mai J. Sci. 2014; 41(3) 593

2.7 Ethnopharmacological UsesThe aerial parts of this plant are used

in traditional medicine in the Caribbeanand Central America against bronchitis,colds, abdominal pains, dysmenorrheal[13], and even as a fertility enhancer [14].In folk medicine, it is employed to treatbackache, muscle cramps, rheumatism,stubborn wounds, sores and swellings, andarthritic painful joints [15]. The MiskitoIndians of eastern Nicaragua use leaves fortreatment of kidney dysfunctioning, cold,stingray wounds, snakebite, purge andamenorrhea [16,17]. Coe and Anderson(1996) reported that fruits, leaves and stemare used in childbirth and in the treatmentof bites and stings, fever and infection [14].W. trilobata, was utilized in Hong Kongas a substitute for W. chinensis, a traditionalChinese medicine used for the treatmentof the common cold, hepatitis, indigestionand infections [18]. In Trinidad andTobago, used for reproductive problems,amenorrhea, dysmenorrheal [19]. It is usedfor the treatment of fever and malaria inVietnam [20].

Unpublished reports indicate thataqueous infusion has been employedlocally and empirically in Southern part ofBrazil in the management of diabetes. Infact, it is popularly referred to as insulinadue to its observed antidiabetic properties[20]. Flowers and leaf part of the plant wereused in the ladies for the purpose ofamenorrhea, childbirth, abortion and toclear the placenta after birth [21,22]. Theliterature review reveals that the freshentire plant is used as molluscicidal activity,antibacterial and antimycobacterialactivity [21].

Suriname’s traditional medicine usesthe stem, leaves, and flower boiled in waterfor hepatitis, indigestion due to sluggishliver, white stools, burning in the urine and

stopping of urine and for infections. Boiledfresh stems and leaves were used for bathethose suffering from backache, musclecramps, rheumatism, or swellings. Used forpainful joints of arthritis, fresh leaves andstems are mashed and spread on a cloth andapplied to area, wrapped securely with awarm covering in South America. Wedeliaspecies is used in lower Thailand forheadache and fever [22].

2.8 Phytochemical ReportsThe main secondary metabolites from

this plant mainly consist of terpenoids,flavonoids and polyacetylenes as well assteroids [20, 23-25]. The leaves and stemcontains eudesmanolide lactones, luteolinand kaurenoic acid [26,27]. It has followingdifferent classes of phytoconstituents.Sesquiterpenoids, triterpenoid andditerpenoid: from the aerial part [18,20, 23-25, 28-34] and the flower of W. trilobata,[18,20, 23-25, 28-35], sterols: [23,25,31],flavonoids: [25], Benzene derivatives: [25,31].

Wedelolactone was reported forhepatoprotective activity, antibacterial,anti hemorrhagic and antiepileptic activity[36-38]. These necessitate estimation ofwedelolactone in Wedelia species forexploring its bioactivity. Wedelolactonewas estimated by high performance thinlayer chromatographic technique. Thestandard wedelolactone showed retentionfactor (Rf) value of 0.56 and this constituentwas found to be present in the testedethanolic extract of W. trilobata. The Rfvalues obtained for wedelolactone in theextracts was found to be 0.56. The amountof wedelolactone was estimated by comparingthe peak area of standard and that presentin the ethanolic extract. The content ofwedelolactone present in the extract wasfound to be 0.084% w/w [21].

594 Chiang Mai J. Sci. 2014; 41(3)

Essential oils are valuable plant productsgenerally of complex compositioncomprising the volatile principles containedin the plant and more or less modifiedduring the preparation process. Mostconstituents of oil belong to the large groupof terpenes. The essential oil obtained fromthe leaves of W. trilobata was analyzed byGC/MS. α- pinene (above 30%), α-phellandrene (17.4%) and limonene (16.3%)were the major components [39].

The fourteen volatile componentswere identified from the essential oils ofW. trilobata leaves, stem and flowers. Theessential oil was characterized by a highpercentage of hydrocarbon sesquiterpenes(HS) (25.5-86.4%), hydrocarbon monoterpenes(HM) (22.9-72.3%) and low levels ofoxygenated sesquiterpenes (OS) (0.0- 7.4%).The major components of volatile oils weregermacrene D (11.9-35.8%), α-phellandrene(1.4-28.5%), α-pinene (7.3-23.8%),E-caryophyllene (4.6-19.0%), bicycloger-macrene (6.0-17.0%), limonene (1.8-15.1%)and α-humulene (4.0-11.6%). The contentof the monoterpenes α-pinene, α-phellandrene and limonene increased fromthe mid-rainy season until the middle ofthe next dry season [40]. Table 1 shows thechemical structures of phytoconstituentpresent in W. trilobata.

2.9 Pharmacological ActivitiesTable 2 summarizes the pharmacological

activity of W. trilobata.

Antidiabetic ActivityMale albino rats with diabetes induced

by the administration of streptozotocin(45 mg/kg, i.v.) were treated with oraladministration of W. trilobata (50 mg/kg).It was found to reduce blood glucose levelsand improved weight gained which wasaccompanied by a marked restoration of

decreased vitamin C and reducedglutathione in liver and kidney tissues ofSTZ-treated rats. In vitro data revealed thatW. trilobata caused an inhibition of lipidperoxidation under Fe2+ or sodiumnitroprusside assaults. Conversely, W.trilobata also caused a reduction in the highlevels of thiobarbituric acid reactivesubstances (TBARS) observed in the liver,kidney, and testes as well as high serumtriglyceride, ALT and AST of diabetic rats[41]. Rungprom et al. (2010) demonstratedthat the methanolic extract of W. trilobatawas found to be the potent α-glucosidaseinhibitor comparable to the authentic drug,Acarbose® [42].Table 1. Secondary metabolites of Wedeliatrilobata (L.)

Compound name Structure Plant part References

Diterpenes Aerial part 25, 32-35ent-kaur-16-en-19 oicacid (Kaurenoic acid)

ent-kaura-9(11), 16- Aerial part 25, 31, 33dien-19 oic acid(Grandiflorenic acid) leaves 35

ent-kaura-9(11), 16- Aerial part 25dien-19 oic acidmethyl ester

3α- (senecioyloxy)- Aerial part 32ent-kaur-16-en-19 oicacid

(3α)-3-(angeloyloxy)- Aerial part 23-25, 32ent-kaur-16-en-19 oicacid

3α-(angeloyloxy) 9β- Aerial part 23hydroxy-ent-kaur-16-en-19 oic acidMethyl-3α-(angeloyloxy) 9β- Aerial part 23hydroxy-ent-kaurenoate

(3α)-3-(tiglinoyloxy)- Aerial part 23-25ent-kaur-16-en-19 oicacid

(3α)-3- Aerial part 23-25, 32(cinnamoyloxy)-ent-kaur-16-en-19 oic acid

(3α)- (cinnamoyloxy)- Aerial part 239βhydroxy-ent-kaur-16-en-19 oic acidMethyl-3α-(cinnamoyloxy)-9β- Aerial part 23hydroxy-ent-kaurenoate

Chiang Mai J. Sci. 2014; 41(3) 595

Table 1. Continued.Compound name Structure Plant part References

15α-(cinnamoyloxy)- Aerial part 32ent-kaur-16-en-19 oicacid(3α)-3-(cinnamoyloxy)-17- Aerial part 25hydroxy-ent-kaur-15-en-19 oic acid(Wedelidin A)(3α)-3-(cinnamoyloxy)-17- Aerial part 25oxo-ent-kaur-15-en-19oic acid (Wedelidin B)

Wedelia-seco- Aerial part 23, 24kaurenolide

Triterpenes

Friedelinol Aerial part 25

Friedelin Aerial part 25

Sesquiterpenes1β, 4α-dihydroxy-6β-isobutyryloxy-9α- Aerial part 29(tigloyloxy)prostatolide9α-(angeloyloxy)-1β,4α-dihydroxy-6 β- Whole plant 29isobutyryloxyprostatolide1β, 9α-diacetoxy-4α-hydroxy-6 β- Whole plant 29isobutyryloxyprostatolide1β, 9α-diacetoxy-4α-hydroxy-6 β- Whole plant 29methacryloxyprostatolide

Wedeliatrilolactone B Aerial part 25

Trilobolide 6-O- Flower 23isobutyrate Leaves 28, 29

Whole plant 18, 20,30

Trilobolide 6-O- leaves 23angelate

Trilobolide 6-O- leaves 20, 23methacrylate

Oxidoisotrilobolide 6- leaves 23, 30O-isobutyrate

Oxidoisotrilobolide 6- leaves 23O-angelate

Oxidoisotrilobolide 6- leaves 23O-methacrylate

Wedelolide A Leaves 20

Table 1. Continued.Compound name Structure Plant part References

Wedelolide B Leaves 20

Wedelolactone Entire plant 21

Ivalin Aerial part 25

Germacrene D Aerial part 23

α-humulene Aerial part 23, 40

Caryophyllene Aerial part 23, 40

SteroidsStigmasterol Aerial part 25

(7α)-7- Aerial part 25hydroxystigmasterol

(3β)-3-hydroxy Aerial part 25stigmasta-5, 22-dien-7-one

Sitosterol Aerial part 23, 40

Daucosterol Aerial part 31

Squalene Aerial part 23, 40

Flavonoids3-hydroxy-6- Aerial part 25methoxychromen-4-one

Apigenin Aerial part 25

Diosmentin Aerial part 25

Benzene derivativesBenzeneacetic acid 2- Aerial part 25phenylethenyl ester

Isocinnamic acid Aerial part 25

4-methoxy catechol Aerial part 25

596 Chiang Mai J. Sci. 2014; 41(3)

Table 1. Continued.Compound name Structure Plant part References

p-cymene Aerial part 23, 40

Protocatechualdehyde Aerial part 31

Caffeic acid Aerial part 31

Cyclic terpenesα-phellandrene leaves 39, 40

α-pinene leaves 40

d-limonene leaves 39, 40

Bicyclogermacrene leaves 40

Antileishmaniasis activityKaurenic acid (ent-kaur-16-in-19-oic),

isolated from the Venezuelan plant W. trilobatawas evaluated on Leishmania (V) braziliensisboth in vivo and in vitro. The compoundhad a lethal effect on axenic amastigotes andpromastigotes with LD50 of 0.25 and 0.78μg/ml, respectively, in 24 h. Additionally,a 70% reduction was observed in the size ofthe skin lesions in Balb/c mice with no evidenttoxic effect. The results indicated that thiscompound has a potent leishmanicidaleffect on L. (V.) braziliensis [34].

Extract/compound Pharmacological activity Reference

Table 2. Biological and pharmacological activities (in vitro and in vivo) of W. trilobata.

ANTIBACTERIAL ACTIVITYN-hexane extract of aerial Inhibitory effect on Gram positive bacteria, 44part without flower Bacillus cereus, Bacillus subtilis, Mycobacterium

megmatis, Staphylococcus aureus, Staphylococcusepidermidis and Gram negative bacteria, Proteusvulgaris, Pseudomonas aeruginosa, Salmonella group C,Salmonella paratyphi, Shigella sonnei

Ethyl acetate extract of Inhibitory effect on Gram negative bacteria, 44aerial part without flower Salmonella group C

Ethanol extract of stem Inhibitory effect on Bacillus subtilis, Pseudomonas 27fluorescens, Clavibacter michiganensis sub sp.michiganensis, Xanthomonas oryzae pv. oryzae,Xanthomonas axanopodis pv. malvacearum andstrains of Staphylococcus aureus, Escherichia coli,Pseudomonas aeruginosa and Klebsiella pneumonia

Ethanol extract of leaves Strong inhibitory effect on P. aeruginosa, K. 27pneumoniae, P. fluorescens, X. oryzae pv. oryzae, X.axanopodis pv. malvacearum, moderately inhibitedthe E.coli, C. michiganensis sub sp. michiganensis butless activity was observed on S. aureus.

Ethanol extract of flower Strong inhibitory effect on Staphylococcus aureus, 27X. oryzae pv. oryzae moderately inhibited theK.pneumoniae, P. fluorescens, X. axanopodis pv.malvacearum but less activity was observed onE.coli, P. aeruginosa, Clavibacter michiganensissub sp. Michiganensis

Chiang Mai J. Sci. 2014; 41(3) 597

Extract/compound Pharmacological activity Reference

Table 2. Continued.

Methanol extract of flower Moderate inhibitory activity against Bacillus cereus, 46Bacillus subtilis, Escherichia coli, Klebsiella pneumonia,Staphylococcus aureus and Shigella flexneri

ANTIFUNGAL ACTIVITYEthanol extract of stem, Weak inhibition against Aspergillus flavus 27leaves and flower A. niger, A. nidulans, A. Flaviceps, Fusarium solani,

F. oxysporum, F. verticilloides

ANTIOXIDANT ACTIVITYEthanol extract of leaf, DPPH radical scavenging activity was more for leaves 27stem and flower than stem and flower

Ethyl acetate fraction of DPPH radical scavenging activity 40Wedelia trilobata

Methanol extract of flower DPPH radical scavenging activity 46

ANTIINFLAMMATORY ACTIVITYEthanol extract of leaf, All the three extract was effective in inhibiting heat 27stem and flower induced albumin denaturation. Maximum inhibition

87.14% was observed from leaf extract followed bystem (86.76%) and flower (61.63%).

The extracts inhibited the heat induced hemolysis of 27RBCs to varying degree. The maximum inhibitions78.11% from leaf extract followed by stem (74.17%)and flower (58.74%).

The ethanolic extract exhibited significant 27antiproteinase activity from different parts. Themaximum inhibition was observed from leaf ethanolicextract (84.19%), in decreasing order was stem(81.84%) and flower ethanolic extract (67.17%).

WOUND HEALING ACTIVITYEthyl acetate (WEA) and The WEA displayed antibacterial and fibroblast 47chloroform:methanol (50:50) stimulatory activities while WCM exhibited antioxidant(WCM) fractions from activityethanolic extract ofW. trilobata leaves

ent-kaura-9(11), 16-dien-19- Offered wound healing activity due to a combination 35oic acid isolated from of antimicrobial, stimulation of fibroblast growthW. trilobata leaves

CNS DEPRESSANT ACTIVITYThe petroleum ether, The petroleum ether extract represented good 43chloroform, ethyl acetate and CNS depressant activitymethanol extract of leaves

CYTOTOXIC ACTIVITYn-hexane and ethyl alcohol The ethyl alcohol extracts of flower had good 45extracts anti-migration and anti-invasion ability especially

on 80 μg/mL dose

598 Chiang Mai J. Sci. 2014; 41(3)

Extract/compound Pharmacological activity Reference

Table 2. Continued.

ANALGESIC ACTIVITYEthanol extracts Blocked the writhing response by 49.17% 48

Kaurenoic acid, a diterpene Exhibited analgesic effect by inhibiting cytokine 49obtained from Wedelia production and activation of the NO-cyclictrilobata GMP-protein kinase G-ATP sensitive potassium

channel signaling pathway.

ANTILEISHMANIASIS ACTIVITYKaurenoic acid (ent-kaur- Potent leishmanicidal effect on L. (V.) braziliensis 3416-in-19-oic), isolated fromthe Venezuelan plantW. trilobata

ANTIDIABETIC ACTIVITYAqueous extract of leaves Reduction in blood glucose level in streptozotocin 41

induced diabetes

Methanolic extract of aerial α-glucosidase inhibitor 42parts

Central nervous system (CNS) depressantactivity

The petroleum ether, chloroform, ethylacetate and methanol extract of leaves of W.trilobata (30 mg/kg, i.p.) were evaluated forCNS depressant activity using pentobarbitone-induced sleeping time, and locomotor activityin mice. The petroleum ether extractpotentiated pentobarbitone sodium inducedsleeping time in mice than other extracts. Theanimal treated with petroleum ether extractshowed reduction in the locomotor activityscores was significantly higher than that ofstandard drug diazepam and other extract.The petroleum ether extract representedgood CNS depressant activity [43].

Antimicrobial activityA biological screening of activity

against Gram-positive and Gram-negativebacteria, yeasts, and fungi of crude extractsfrom W. trilobata (10 μg/ml) was reported.The N-hexane extract showed antibacterial

activity against Bacillus subtilis, Mycobacteriumsmegmatis, Staphylococcus aureus, andStaphylococcus epidermidis (Gram-positivebacteria); along with Proteus vulgaris,Pseudomonas aeruginosa, Salmonella groupC, Salmonella paratyphi, and Shigella sonnei(Gram-negative bacteria). The ethyl acetateextract was active only against Salmonellagroup C; and the aqueous extract wasinactive against the tested bacteria. Noneof the tested extracts showed biologicalactivity against the yeasts (Candida albicans,Candida tropicalis, Rhodotorula rubra) or thefungi (Aspergillus flavus, Aspergillus niger,Mucor sp., Trichophyton rubrum) [44].

Ethanol extract of leaf, stem and flowerof W. trilobata (10 μg/ml) was assessed forits antimicrobial efficacy using disc methodagainst different fungi (A. flavus, A. niger,A. nidulans, A. flaviceps, Alternaria carthami,Alternaria helianthi, Cercospora carthami,Fusarium solani, Fusarium oxysporum,Fusarium verticilloides and Nigrospora oryzae)

Chiang Mai J. Sci. 2014; 41(3) 599

and bacteria (B. subtilis, Pseudomonasfluorescens, Clavibacter michiganensis subsp. michiganensis, Xanthomonas oryzae pv.oryzae, Xanthomonas axanopodis pv.malvacearum and strains of S. aureus, E. coli,P. aeruginosa and K. pneumoniae). Theethanolic stem extract significantly inhibitedthe growth of almost all the bacteria isolatesbut did not show any significant effect onfungal isolates. The leaf extract showed morepotent against P. aeruginosa, K. pneumoniae,P. fluorescens, X. oryzae pv. oryzae, X.axanopodis pv. Malvacearum [27].

W. trilobata flowers, leaves and stemswere extracted with ten times of ethyl alcohol.The extract was then partitioned by N-hexane, ethyl acetate, N-butyl alcohol andwater to evaluate its antimicrobial activity.The result showed that most extracts hadantimicrobial activities except the waterextracts from flower. The ethyl acetateextract was the most effective among all theextracts [45].

The methanolic flower extract of W.trilobata was screened for antibacterialactivity by disc diffusion method againstBacillus cereus, Bacillus subtilis, Escherichiacoli, Klebsiella pneumonia, Staphylococcusaureus and Shigella flexneri. The extractshowed a moderate inhibitory activityagainst all bacterial species with zones ofinhibition of 10-16 mm in comparison withchloramphenicol which showed zones ofinhibition 12-24 mm [46].

Antioxidant activityEthanol extract of leaf, stem and flower

(0.5 mg/ml) of W. trilobata was evaluatedfor its antioxidant activity by measuringthe scavenging activity of 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical and theferric reducing antioxidant power (FRAP)assay. It was observed that ethanol extractof the leaf of W. trilobata offered higher

activity than that of stem and flower. At aconcentration of 0.1 mg/ml, the scavengingactivity of ethanol extract of the stem andflower and leaves reached 82.64, 55.41 and86.17% respectively but less than those ofascorbic acid (98%) and BHT (97.8%) at 0.1mg/ml, the study showed that the extractshave the proton donating ability and couldserve as free radical inhibitors orscavenging, acting possibly as primaryantioxidants. The FRAP values for theethanol extract of leaf and stem weresignificantly lower than that of ascorbicacid but higher than that of BHT [27].

The methanolic extract of W. trilobataflower showed good antioxidant activity(IC50 = 90 μ g/ml) in DPPH method.Reference standard ascorbic acid showedIC50 of 60 μg/ml. The extract of W. trilobataflower exhibited higher ABTS (2,2’-azino-bis (3- ethylbenzothiazoline-6- sulphonicacid) radical scavenging activity with IC50

of 80 μg/ml whereas gallic acid showedIC50 of 30 μg/ml [46].

Anti-inflammatory activityEthanol extract of leaf, stem and flower

(0.5 mg/ml) of W. trilobata was evaluatedfor its in vitro anti-inflammatory usingalbumin denaturation, membrane stabiliza-tion assay and proteinase inhibitory assay.All the three extracts were effective ininhibiting heat induced albumin denaturation.Maximum inhibition 87.14% was observedfrom leaf extract followed by stem (86.76%)and flower (61.63%). All the extracts wereeffective in inhibiting the heat inducedhemolysis. The extracts inhibited the heatinduced hemolysis of RBCs to varyingdegree. The maximum inhibitions 78.11%from the leaf extract followed by thestem (74.17%) and flower (58.74%).The W. trilobata ethanolic extract exhibitedsignificant antiproteinase activity from

600 Chiang Mai J. Sci. 2014; 41(3)

different parts. The maximum inhibitionwas observed from leaf ethanolic extract(84.19%), in decreasing order was stem(81.84%) and flower ethanolic extract(67.17%) [27].

The ethyl alcohol and water extractsof W. trilobata flowers were used to treatRAW 264.7 macrophage, which inducedinflammation by LPS. In the nitric oxideassay, the extracts of W. trilobata flowerhad better inhibitory ability against LPSinduced inflammation [45].

Wound healing activityAn ethanolic extract of W. trilobata

leaves was subjected to column chromato-graphy. Hexane, ethyl acetate (WEA) andchloroform:methanol (50:50) (WCM)fractions were obtained. The fractions weretested using relevant in vitro wound healingassays. WEA (3 μg/mL) promoted fibroblastL929 viability up to more than 90% beforeand more than 85% after hydrogen peroxideinduced oxidative stress. WEA (3 μg/mL)induced a 70% migration rate in the in vitroscratch assay and the collagen content wasincreased to 261 μg/mL compared to thecontrol (57.5 μg/mL) [47].

The ent-kaura-9(11), 16-dien-19-oic acidisolated from W. trilobata leaves offeredwound healing activity due to a combinationof antimicrobial, stimulation of fibroblastgrowth and protection of the cells fromhydrogen peroxide-induced injury, all ofwhich could play some role in its effecton tissue repair. It showed promisingantibacterial activity with MIC value of15.62 μg/mL against S. aureus and 7.81μ g/mL against S. epidermidis. Theent-kaura-9(11), 16-dien-19-oic acid (2.5-0.08μ g/mL) produced an increase in thepercentage viability of mouse fibroblastL929 cells from 97-117% and protection ofthe fibroblast L929 cells against oxidative

stress induced by hydrogen peroxide(94-80%) [35].

Cytotoxic activityIn transient transfection assay the

N-hexane and ethyl alcohol extracts ofW. trilobata flower could activate PPARα.In MTT assay of SK-Hep-1, extract ofW. trilobata flower had the best inhibitoryability. The ethyl alcohol extract of W.trilobata had the best ability to diminishthe expression of matrix metalloproteinase(MMP)-9 and MMP-2. The ethyl alcoholextracts of flower had good anti-migrationand anti-invasion ability especially on 80μg/mL dose [45].

Analgesic activityComparative study in mice on the

analgesic activity of the ethanol extracts ofW. trilobata, W. biflora and E. alba wasevaluated by acetic acid induced writhingmethod. It was found that W. trilobataextract showed dose-dependent blocking ofwrithing response. Dose of 500 mg/kg ofW. trilobata extract and aspirin (500 mg/kg) block the writhing response by 49.17%and 68.68% [48].

Kaurenoic acid (10 mg/kg) obtainedfrom Wedelia trilobata kaurenoic acidinhibited overt nociception like behaviorinduced by phenyl-p-benzoquinone,complete Freund’s adjuvant (CFA) andformalin. Kaurenoic acid (1-10 mg/kg p.o.)also inhibited acute carrageenin and PGE2

induced and chronic CFA inducedmechanical hyperalgesia. [49].

Reproductive problemsPrevious research has shown that

Caribbean women and Creoles have alwaysused bitter herbs to control their fertility.A study was conducted focused on theplants used for reproductive purposes in

Chiang Mai J. Sci. 2014; 41(3) 601

Trinidad and Tobago. The plants used toaddress women’s reproductive problemswere used mainly for infertility, menstrualpain and childbirth. Results showed thatW. trilobata was used for menstrual painand for the female complaints. The non-experimental validation method can beused to advise the public on which plantsare safe, effective and useful, and which arenot; pending clinical trials. This isimportant since few clinical trials wereconducted on Caribbean plants [19].

Other activityThe methanolic extract of W. trilobata

flower was tested for pTZ57R/T plasmidDNA protection against hydroxyl radical,evidenced by fragmentation assay. Theextract showed stronger protective effectagainst DNA damage by hydroxyl radicalreleased by Fenton’s reaction [46].

Toxicity studiesThe ethanolic extract (25- 1.56 μg /ml),

ethyl acetate fraction (6.25- 0.39 μg/ml) andchloroform: methanol fraction (22- 0.39μ g/ml) of leaves of W. triolbata wereevaluated for cytotoxicity using L929mouse fibroblast cell using MTT (3-(4,5-dimethylthiazolyl)-, 5 diphenyltetrazoliumbromide) assay. At these concentrationsextract as well as fractions produced cellsurvivability of more than 80%. Howeverat higher concentrations of the abovedose levels there was evidence ofcytotoxicity [47].

The ent-kaura-9(11), 16-dien-19-oic acidat dose range of 10- 0.08 μg/ml producedcell viability of L929 mouse fibroblast inthe range of 89-117%. Further increase inconcentration above 10 μg/ml producedcellular damage resulting in cell death [35].

The J774-G8 macrophages were treatedwith kaurenoic acid isolated from W.

trilobata at a concentration varyingbetween 10- 250 μg/ml. The effect of thecompound on cellular viability wasevaluated using MTT assay Low toxicitywas observed for J774-G8 macrophageswith a LD50 of 25 μg/ml and high viability(70-92%), while a moderate viability wasobserved for infected macrophages(37-81%), with concentrations of 25 μg/ml or less [34].

3. DISCUSSIONThe present review emphasizes the

phytochemical, traditional, pharmacologicaland, clinical reports on W. trilobata.Tannin, saponins, flavonoids, phenolic,terpenoids constitute major classes ofphytoconstituents of this plant. The plantcontains a range of phytochemical substancescredited with various pharmacologicalproperties. Recent research carried outindicates its uses such as antioxidant, anti-inflammatory, antimicrobial, wound healing,antidiabetic activity. In recent years,the search for phytochemicals possessingantioxidant, antimicrobial and antiinflammatory properties have been on therise due to their potential use in the therapyof various chronic and infectious diseases.Epidemiology and experimental studieshave implicated oxidative cellular damagearising from an imbalance between freeradical generating and scavenging systemsas the primary cause of cardio-vascular,diseases, cancer, aging etc. Due to risk ofadverse effects encountered with the use ofsynthetic antibiotics, medicinal plantsmay offer an alternative source forantimicrobial agent with significant activityagainst pathogenic and infective micro-organisms. In addition, a number ofantibiotics have lost their effectiveness dueto the development of resistant strains,mostly through the expression of resistance

602 Chiang Mai J. Sci. 2014; 41(3)

genes. Strong antioxidants, antimicrobialand anti inflammatory activities specificallyin the ethanolic leaf and stem extracts ofW. trilobata were observed. These activitiesmay be due to strong occurrence ofpolyphenolic compounds such as flavonoids,tannins, terpenoids, phenols and saponins[29]. The authors are involved in evaluatingthe wound healing effect of W. trilobatawith a view to isolating bioactivephytoconstituent(s). One of thephytoconstituent isolated and evaluated forwound healing potential is grandiflorenicacid (ent- kaura-9(11)-dien-19-oic acid) [37,55]. The presence of a wide range ofchemical compounds indicates that the plantcould lead the way for the development ofnovel agents having good biologicalactivity. Exploration of the chemicalcompounds of the plant will provide thebasis for developing such a lead. Thephytomedicines can be developed as analternative and are relatively inexpensivethan modern drugs. One of the reasons istheir use, preparation, and safety is alreadyunderstood in traditional systems ofmedicines [57]. Many chemical compoundsare present in the plant but isolation ofactive constituents can be carried out usingdifferent extraction methods such asmicrowave extraction, isolation and byusing various appropriate chromatographictechniques. Despite a long tradition of use ofW. trilobata for treatment of variousailments, it still remains unexploredpharmacologically to prove its traditionalclaims. Thus it can be considered as avaluable plant in both traditional andmodern drug development areas for itsversatile medicinal uses. Emphasis must belaid on the pharmacological activity of thephytoconstituent to unravel the hiddenmedicinal qualities of this plant as well as

the local knowledge system should beglobalize which would increase the benefitsobtained to wider population. There areno clinical data available that wouldprovide evidence of efficacy of W. trilobatain humans. Extracts and constituents of W.trilobata may have considerable clinicalpotential in humans and need to be studiedfurther in in vivo models and ultimatelyin clinical studies.

DISCLOSURE STATEMENTNo competing financial interests exist.

REFERENCES

[1] Ekpo M., Mbagwu H., Jackson C. andEno M., Antimicrobial and woundhealing activities of Centrosemapubescens (Leguminosae), J. Phys.Chem. Solids, 2011; 1: 1-6.

[2] Bazzaz B.S.F., Arab A., Emami S.A.,Asili J., Hasanzadeh-Khayyat M. andSahebkar A., Antimicrobial andantioxidant activities of methanol,dichloromethane and ethyl acetateextracts of Scutellaria lindbergiiRech.f., Chiang Mai J. Sci. 2013; 40:49-59.

[3] Pradhan D., Panda P.K. and Tripathy G.,Wound healing activity of aqueous andmethanolic bark extracts of Vernoniaarborea Buch-Ham. in Wistar rats,Nat. Prod. Rad., 2009; 8: 6-11.

[4] Joseph B.S. and Justin R.S., Acomparative study on variousproperties of five medicinallyimportant plants, Int. J. Pharmacol.,2011; 7: 206-211.

[5] Clare B.A., Conroy R.S. and Spelman K.,The diuretic effect in human subjectsof an extract of Taraxacum officinalefolium over a single day, J. Altern.Complement. Med., 2009; 15: 929-934.

Chiang Mai J. Sci. 2014; 41(3) 603

[6] Adetutu A., Morgan W.A. andCorcoran O., Ethnopharmacologicalsurvey and in vitro evaluation ofwound-healing plants used in South-western Nigeria, J. Ethnopharmacol.,2011; 137: 50-56.

[7] Thamam R.R., Wedelia trilobata:Daisy invader of the Pacific islands, IAStechnical report 99/2. Institute ofApplied Science, University of SouthPacific, Suva, Fiji, 1999: 1-10.

[8] PIER (Pacific Island Ecosystems atRisk), Wedelia trilobata, 2003. Onlinedocument at: http://www.hear.org/pier/species/species/wedelia_trilobata.htm. (Accessed April 10, 2012).

[9] USDA-GRIN, Sphagneticola trilobata(L.) Pruski. GRIN Taxonmy forPlants. U.S. Department of AgricultureGermPlasm Resources InformationNetwork. Mem, New York Bot,Gard., 1996. Online document at:http://www.ars-grin.gov/cgi-bin/npgs/html/taxon.pl. (Accessed May16, 2012).

[10] USDA-NRCS, The plant database,version 3.5. National plant data center,Baton Rouge, USA, 2002. Onlinedocument at: http://plants.usda.gov.(Accessed May 16, 2012).

[11] Whistler W.A., Wayside Plants of theIslands: A Guide to the Lowland Floraof the Pacifiic Islands including Hawai’i,Samoa, Tonga, Tahiti, Fiji, Guam andBelau: Isle Botanica; Honolulu, 1995.

[12] Wagner W.L., Herbst D.R. andSohmer S.H., Manual of the FloweringPlants of Hawaii. Vol. 2. Honolulu:University of Hawaii Press and BishopMuseum Press, 1990.

[13] Germos n-Robenau L., Hacia unaFarmacopea Cariben~a, Tramil 7, SantoDomingo-Rep blica Dominicana,1995.

[14] Coe F.G. and Anderson G.J., Screeningof medicinal plants used by the Garifunaof eastern Nicaragua for bioactivecompounds, J. Ethnopharmacol., 1996;53: 29-50.

[15] Arvigo R. and Balik M., RainforestRemedies, One Hundred Healing Herbsof Belize, USA: Lotus Press, TwinLakes, WI, 1993.

[16] Barrett B., Medicinal plants ofNicaragua’s Atlantic coast, USA: Econ.Bot., 1994; 48: 8-20.

[17] Duke J., American agriculturalresearch service-phytochemical andethnobotanical Database, 2002. Onlinedocument at: http://www.ars-grin.gov/duke/index.html. (AccessedFebruary 10, 2012).

[18] Huang X.S., Jiang R.W. and Ooi V.,Trilobolide-6-O-isobutyrate, a eudes-manolide from Wedelia trilobata, ActaCrystallogr., 2003; E59: 771–772.

[19] Lans C., Ethnomedicines used inTrinidad and Tobago for reproductiveproblems, J. Ethnobiol. Ethnomed.,2007; 3: 13-25.

[20] That Q.T, Jossang J., Jossang A., KimP.P. and Jaureguiberry G., WedelolidesA and B: Novel sesquiterpene -lactones,(9R)-eudesman-9, 12-olides, from Wedeliatrilobata, J. Org. Chem., 2007; 72:7102–7105.

[21] Shanmugam S., Malligeswari K.S.,Venugopalan R. and Shanmugam T.Estimation of Wedelolactone contentin Wedelia species by HPTLCtechnique, J. Pharm. Res., 2011; 4: 193-194.

[22] Meena A.K., Rao M.M., Meena R.P.,Panda P. and Renu, Pharmacologicaland phytochemical evidences for theplants of Wedelia genus- A review,Asian J. Pharm. Res. 2011; 1: 7-12.

604 Chiang Mai J. Sci. 2014; 41(3)

[23] Bohlmann F., Ziesche J., King R.M.and Robinson H., Eudesmanolidesand diterpenes from Wedelia trilobataand an ent-kaurenic acid derivativefrom Aspilia parvifolia, Phytochem.,1981; 20: 751–756.

[24] Duke J.A., Handbook of PhytochemicalConstituents of GRAS Herbs andOther Economic Plants, Boca Raton,FL: CRC Press, 1992.

[25] Qiang Y., Dub D.L., Chenc Y.J. andGao K., ent-Kaurane diterpenes andfurther constituents from Wedeliatrilobata, Hel. Chim. Acta, 2011; 94:817-823.

[26] Block L.C., Santos A.R.S., Marcia deSouza M., Scheidt C., Yunes R.A., SantosM.A., Monache F.D. and Filho V.C.,Chemical and pharmacological examina-tion of antinociceptive constituents ofWedelia paludosa, J. Ethnopharmacol.,1998; 61: 85-89.

[27] Govindappa M., Naga S.S., PoojashriM.N., Sadananda T.S. and ChandrappaC.P., Antimicrobial, antioxidant andin vivo anti-inflammatory activity ofethanol extract and active phytochemicalscreening of Wedelia trilobata (L.)Hitchc, J. Pharmacog. Phytother., 2011;3: 43-51.

[28] Farag S.F., El-Emary N.A. and NiwaM., Trilobolide-6-O-isobutyrate, aeudesmanolide from Wedelia trilobata,Chem. Pharm. Bull., 1996; 44: 661-664.

[29] Zhang Y.H., Liu M.F., Ling T.J. andWei X.Y., Allelopathic sesquiterpenelactones from Wedelia trilobata, J.Trop. Subtrop. Bot., 2004; 12: 533-537.

[30] Huang X., Ou S., Tang S., Fu L. andWu J., Simultaneous determination oftrilobolide-6-O-isobutyrates A and B inWedelia trilobata by gaschromatography, Chin. J. Chromatogr.,2006; 24: 499-502.

[31] Wu M.L., Zhang Z.D., Xu Q.J., XieR.R. and Li Q.Q., Chemicalconstituents of Wedelia trilobata,Chin. Tradit. Herb Drugs, 2010; 41:681-685.

[32] Bohlmann F. and Van N.L., Naturallyoccurring terpene derivatives Part 97,new kaurenic acid derivatives fromWedelia species, Phytochem., 1977; 16:579-581.

[33] Batista R., Chiari E. and Oliveira A.B.,Trypanosomicidal kaurane diterpenesfrom Wedelia paludosa, Planta Med.,1999; 65: 283-284.

[34] Brito S., Crescente O., Fern ndez A.,Coronado A. and Rodriguez N., Efficacyof a kaurenic acid extracted from theVenezuelan plant Wedelia trilobata(Asteracea) against Leishmania (Viannia)braziliensis, Biomed., 2006; 26: 180-87.

[35] Balekar N., Nakpheng T., KatkamN.G., Srichana T. Wound healingactivity of ent-kaura-9(11), 16-dien-19-oic acid isolated from Wedeliatrilobata (L.) leaves, Phytomed., 2012;19: 1178-1184.

[36] Murugaian P., Ramamurthy V. andKarmegan N., Hepatoprotective activityof Wedelia calendulaceae L. againstacute hepatotoxicity in rats, Res. J.Agric. Biol. Sci., 2008; 4: 685-687.

[37] Upadhyay K., Gupta N.K. and DixitV.K., Development and characterizationof phyto-vesicles of wedelolactone forhepatoprotective activity, Drug Dev.Ind. Pharm., 2012; 38: 1152-1158.

[38] Shaikh M.F., Sancheti J. and SathayeS., Phytochemical and pharmacologicalinvestigations of Eclipta alba (linn.)hassak leaves for antiepileptic activity,Int. J. Pharm. Sci., 2012; 4: 319-323.

Chiang Mai J. Sci. 2014; 41(3) 605

[39] Craveiro A.A., Matos F.J.A., AlencarJ.W., Machado M.I.L., Krush A. andSilva M.G.V., Volatile constituents oftwo Wedelia species, J. Essent. Oil Res.,1993; 5: 439-441.

[40] da Silva C.J., Barbosa L.C.A., DemunerA.J., Montanari R.M., Francino D.,Meira R.M.S.A. and de souza A.O.,Chemical composition and histochemis-try of Sphagneticola trilobata essentialoil, Braz. J. Pharmacog., 2012; 22: 1-8.

[41] Kade I.J., Barbosa N.B.V., IbukunE.O., Igbakinc A.P., Nogueirab C.W.and Rochab J.B.T., Aqueous extractsof Sphagneticola trilobata attenuatesstreptozotocin-induced hyperglycaemiain rat models by modulating oxidativestress parameters, Biol. Med., 2010; 2:1-13.

[42] Rungprom W., Siripornvisal S.,Keawsawai S. and Songtrai M.,α-Glucosidase inhibitors from medicinalplants for diabetes therapy, Agric. Sci.J., 2010; 41: 301-304.

[43] Tambe V.D., Nirmal S.A. and JadhavR.S., Central nervous system depressantactivity of Wedelia trilobata leaves, DhakaUniv. J. Pharm. Sci., 2007; 6: 59-60.

[44] Taddei A. and Rosas-Romero A.J.,Antimicrobial activity of Wedelia trilobatacrude extracts, Phytomed., 1999; 6: 133-134.

[45] Wei L.C., Studies on the antibacterialactivity, antioxidation, anti-inflammation

and anticancer activity of Wedeliatrilobata (L.) Hitchc. 2009. Availableat: http://asiair.asia.edu.tw/handle/310904400/2920. (Accessed February12, 2012).

[46] Chethan J., Sampath Kumara K.K.,Niranjana S.R. and Prakash H.S.,Evaluation of antioxidant and anti-bacterial activities of methanolic flowerextract of Wedelia trilobata (L.) Hitch.Afr. J. Biotechnol., 2012; 11: 9829-9834.

[47] Balekar N., Katkam N.G., NakphengT., Jehtae K. and Srichana T.,Evaluation of the wound healingpotential of Wedelia trilobata (L.)leaves, J. Ethnopharmacol, 2012; 141:817-24.

[48] Sureshkumar S., Bhama S., Siva KumarT., Chandrasekar M.J.N. and Rajesh R.,Analgesic activities of the medicinalplants of Wedelia trilobata, Wedeliabiflora and Eclipta alba in standardexperimental animal models, Biosci.Biotechnol. Res. Asia, 2007; 4: 201-206.

[49] Mizokami S.S., Arakwa N.S., AmbrosioS.R., Zarpelon A.C., Casagrande R.,Cunha T.M., Frreira S.H., CunhaF.Q. and Verri W.A., Kaurenoic acidfrom inhibits inflammatory pain:Effect on cytokine production andactivation of the NO−Cyclic GMP−protein kinase G−ATP-sensitivepotassium channel signaling pathway,J. Nat. Prod., 2012; 75: 896-904.