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�:Biological Activities of Aristolochia Plants: A Mini Review
Bachir BENARBA1 & Boumedienne MEDDAH2

1Laboratory for Research on Biological Systems and Geomatics, Department of Biology, Faculty of Nature and Life Sciences, University of Mascara, Algeria

2 Laboratoire de Bioconversion, G�nie Microbiologie et S�curit� Sanitaire, Department of Biology, Faculty of Nature and Life Sciences, University of Mascara, Algeria

Address for correspondence: Dr. Bachir Benarba, Laboratory for Research on Biological Systems and Geomatics, Faculty of Nature and Life Sciences, University of Mascara, Algeria. E-mail: bachirsb@yahoo.fr

 Abstract
The genus Aristolochia (Aristolochiaceae) consists of approximately 500 species, most of which are found in tropical, subtropical, and Mediterranean regions. Plants of the genus Aristolochia have been shown to exhibit interesting biological activities including cytotoxic, apoptosis-induced, antimicrobial, anti-inflammatory, antioxidant, antiallergic activities, among other. The present review summarizes some demonstrated biological activities of the plants of the genus Aristolochia reported to date. 

Contents 
Introduction
Ethno medicinal uses of Aristolochia species
Anticancer activity
Antimicrobial activity
Anti-inflammatory activity
Antioxydant activity
Anti-allergic activity
Conclusion
Introduction 
The Aristolochiaceae family is usually divided into two subfamilies: Asaroideae and Aristolochioideae. The former includes Asarum and Saruma, and the latter includes Aristolochia sensu lato (s.l.) and Thottea genera [1].
Aristolochia, Fam. Aristolochiaceae, is a large genus of herbs and twining plants, found in the tropical and temperate regions of the world [2]. In the Mediterranean and adjacent Near East 50 Aristolochia species occur, all of them being endemic to a particular area [3] (figure). 

Figure 1: Distribution of Aristolochiaceae [4]


Ethno medicinal uses of Aristolochia species

Species of Aristolochia have often been reported as important medicinal plants in ethnobotanical studies [5].
Aristolochia species are traditional and folk medicines in China as anti-inflammatory agents for arthritis, gout, rheumatism and dieresis [6, 7]. They have been used in slimming therapy as a substitute for, or in addition to, traditional Chinese plants [8]. It has been reported that A. contorta from China and Korea roots and fruit are used as anticancer drug (they contain lysicamine and oxaaporphine) [9]. A. longa is widely used in anticancer treatment in Algeria [10, 11, 12] and Morocco [13]. 
The stem of A. manshuriensis (AMA, Guanmuton), a traditional Chinese medicinal herb largely harvested from the Northeast of China, is used as a diuretic, anti-inflammatory, to alleviate swelling and to treat rheumatism [14]. Members of this genus are used in Brazilian traditional medicine as a stomachic, anti-inflammatory, antiasthmatic and abortifacient, as well as an antidote for snakebite, to cure several types of cancer [15]. In Brazil, roots of A. birostris Duch. and A. papillaris Mart. are used as abortive and in treatment of amenorrhea [16]. Roots of A. cymbifera Mart. & Zucc. are used for Asthma, fever, diarrhea, dyspepsia, gout, infection, Amenorrhea, orchitis, intermittent fevers [17]. A. elegans is traditionally used to treat malaria [18].

Anticancer activity
Some Aristolochia species were reported to have demonstrated anticancer activities, 
such as A. baetica [19], A. acuminata [20], A. birostris [21], A. constricta [22],  and A. rotunda [23].
It has been reported that extracts of plants belonging to the genus Aristolochia induced growth inhibition of human cell lines in a dose dependent fashion.
We have recently demonstrated that an aqueous extract of A. longa induced cell death of Burkitt�s lymphoma cell line (BL41) in a dose-dependent manner. The IC50 of A. longa aqueous extract was estimated at about 15,63 �g/ml. The extract induced apoptosis in Burkitt�s lymphoma BL41 cells, by triggering the mitochondrial pathway (disruption of ��m, activation of caspases-9 and -3 and PARP cleavage) [24].
A. macroura ( from Argentina) methanolic extract inhibited growth of  Hep G2 cell line in a concentration-dependent manner, the IC50 was estimated to be 513�91 �g/ml [25]. Chaouki et al. (2010) studied the antiproliferative effect of four different polarity extracts from A. baetica L. growing in Morocco on MCF-7 cells. All the extracts showed potent cell proliferation inhibition on MCF-7 cells. The chloroform extract (IC50: 216.06 � 15 �g/ml) was the most active [26]. The in vitro cytotoxicity of the essential oils from the rhizome and the aerial part of A. mollissima on six human cancer cell lines (Human breast cancer cell lines (MCF-7 andMDA-MB-435S), human cervix carcinoma cell line (HeLa), human liver carcinoma cell lines (Bel-7402 and Hep G2) and human renal adenocarcinoma cell line (ACHN)) was investigated. Dichloromethane and methanolic extracts of A. triangularis were cytotoxic to the KB (Human oral epidermoid carcinoma) cells, with IC50 47�12 and 87�22, respectively [27]. The cytotoxicity of the rhizome oil on four cancer cell lines (ACHN, Bel-7402, Hep G2 and HeLa; IC50: 22,3; 33.1�; 33.2�; 38.6 (�g/ml) respectively) were significantly stronger than that of the oil from aerial part of A. mollissima. Among six cancer lines tested, ACHN, MCF-7 and MDA-MB-435S were more sensitive to two oils than other cancer cell lines [28].
Extracts of Aristolochia plants could be a promising source of cytotoxic molecules. Hegde et al. (2010) isolated from the 70% aqueous methanolic extract of the Chinese plant A. manshuriensis two novel substituted phenanthrene compounds (figure). They were identified as inhibitors of the CDK2 enzyme. Compound 1 was found to be a potent inhibitor of the CDK2 enzyme with an IC50 of 140 nM, whereas compound 2 was found to be less active with an IC50 of >10 �M [29].

Figure 2: Inhibitors of the CDK2 enzyme isolated from [29]

Licarin A isolated from Aristolochia pubescens [30] was found to show significant cytotoxic activity against six human cancer cell lines: A-549 (lung), MCF7 (breast), HCT-15 (colon) [31], P-388, KB16 and HT-29 [32]. 

Antiproliferative activity of argentilactone isolated from Aristolochia argentina against 
MCF-7, NCI-ADR (breast expressing the multidrug resistance phenotype), NCI 460 (lung, nonsmall cells), UACC62 (melanoma), 786-0 (kidney), OVCAR03 (ovarian), PCO 3 (prostate), and HT-29 (colon) was demonstrated. 


Figure 3�: Argentilactone isolated from Aristolochia argentina [33]


The natural form of argentilactone [(R)-(6)] proved to be more potent for breast cancer cells that express multidrug resistance phenotype (NCI-ADR) [33].

Antimicrobial activity

Navarro-Garc�a et al. (2011) evaluated the dichloromethane extract from A. brevipes (Rhizoma) and the compounds isolated from this extract against several mycobacterial strains, sensitive, resistant (monoresistant), and clinical isolates (multidrug-resistant). Results of this study demonstrated that the dichloromethane extract (rhizome) of A. brevipes possessed strong in vitro antimycobacterial activity against Mycobacterium tuberculosis H37Rv (Minimum Inhibitory Concentration value [MIC], 12.5 �g/ml). The most active compound against all mycobacterial strains tested was the compound aristolactam I, with MIC values ranging between 12.5 and 25 �g/ml  [34]. The methanol extracts and the hexane fractions of either the rhizome or the leaves of A. paucinervis (Morocco) exhibited an inhibitory activity against Helicobacter pylori at a concentration of d"128 �g/ml [35]. Deffated chloroform fraction obtained from the rhizomes of the plant produced an intense time-dependent bactericidal effect against C. perfringens, achieving over a 24 h- period a 5 log10-unit decrease in CFU/ml at a concentration>1.25xMIC [36]. The hexane extract from stems of A. cymbifera Duch., showed good activity against Staphylococcus spp. and Pseudomonas aeruginosa. The bioactive compound in this fraction was the diterpene 2-oxo-populifolic acid [37]. Camporese et al. (2003) evaluated Twenty-one extracts from seven herbal drugs, A. trilobata leaves and bark, Bursera simaruba (Burseraceae) bark, Guazuma ulmifolia (Sterculiaceae) bark, Hamelia patens (Rubiaceae) leaves and Syngonium podophyllum (Araceae) leaves and bark, used in traditional medicine of Belize (Central America). Activity was tested against standard strains of Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, Staphylococcus aureus ATCC 25923 and Enterococcus faecalis ATCC 29212. They found that the hexane extracts of A. trilobata leaves and bark were the most active extracts against Staphylococcus aureus (MIC = 0.31 and 0.625 mg/ml, respectively) [38]. It has been reported that the chloroform and hexane extracts of A. trilobata leaves and bark were active against Escherichia coli and Pseudomonas aeruginosa and Staphylococcus aureus [39]. Furthermore, extracts of A. bracteata were tested against Gram-positive and Gram-negative (Bacillus subtilis and Escherichia coli) bacteria. Aristolochic acids I isolated from ethyl acetate and methanol extracts were shown good antimicrobial activity and the zone of inhibition of both at higher concentration 50 �g/ml was similar with the standard aristolochic acid [40]. A. longa was reported as as good antimicrobial drug [41]. Aristolochic acids, isolated from this plant inhibited Escherichia coli, Pseudomonas aeruginosa, Streptococcus faecalis, Staphylococcus aureus and Staphylococcus epidermidis [39]. The antimicrobial activity of essential oils from the rhizome and the aerial part of A. mollissima was evaluated against 20 microorganisms using disc diffusion and broth microdilution methods. The gram-positive bacteria were more sensitive to both oils than gram-negative bacteria and yeasts. The rhizome oil showed the strongest bactericidal activity against Staphylococcus saprophyticus, whereas the oil from the aerial part of A. mollissima exerted the strongest bactericidal activity against methicillin-resistant and methicillin-senstive Staphylococcus aureus [42].

Anti-inflammatory activity 

The anti-inflammatory property of Aristolochia species is probably the result of a direct interaction between aristolochic acid and derivatives of phospholipase A2 [43]. Sosa et al. (2002) evaluated Hexane, chloroform and methanol extracts of seven herbal drugs used in the folk medicine of Central America against skin disorders (A. trilobata leaves and bark, Bursera simaruba bark, Hamelia patens leaves, Piper amalago leaves, and Syngonium podophyllum leaves and bark) for their topical anti-inflammatory activity against the Croton oil-induced ear oedema in mice; they found that the most active drug was A. trilobata leaves.  The chloroform extract of A. trilobata leaves showed an anti-inflammatory effect comparable to that of indomethacin: its ID50 value was 108 mg/cm2 whereas that of the reference drug is 93 mg/cm2. They concluded that among the plants used, the most promising one was that of A. trilobata leaves, which antiphlogistic potency is close to that of the pure non-steroidal anti-inflammatory drug indomethacin [44]. Recently, Battu  et al. (2011) investigated (for the first time) the anti-inflammatory activity of the petroleum ether, ethyl acetate, and ethanol extracts of A. tagala roots in vivo (the carrageenan-induced hind paw edema in rats) and in vitro (macrophage cell line RAW264.7 stimulated with proinflammatory stimuli (lipopolysaccharide interferon � or the calcium ionophore A23187)). All the extracts exhibited excellent anti-inflammatory effects demonstrating that that the A. tagala roots had excellent anti-inflammatory activity and validated the traditional indications of this plant [45].

Antioxydant activity 
Important antioxidant activity has been reported for Aristolochia plants. A. cymbifera (rhizomes) alcoholic extract (50% ethanol) showed good antioxidant potential (IC50 = 124.5 � 6.6 using  DPPH photometric assay) [46]. Thirugnanasampandan et al. (2008) evaluated antioxidant properties of five ristolochia species: Aristolochia brasiliensis Mart. And Zucc., Aristolochia bracteolata Retz., ristolochia indica Linn., and Aristolochia tagala Cham. All the extracts (petroleum ether, chloroform, and ethyl acetate extracts) exhibited antioxidant properties. The most effective free radical scavenging activity was shown by all three extracts of A. tagala [47]. Furthermore, phenolic extracts of A. longa growing in Algeria were shown to posses promising antioxidant activity [48].


Antiallergic activity 
It has been reported that some Aristolochia species were used as antiallergic plants. In India, roots of A. indica L are used to treat skin allergies [49]. Chitme et al. (2010) investigated antiallergic activity of A. bracteolata in animal model. It has been demonstrated that chloroform extract of the plant had potent inhibitory effect on 48/80 induced pruritis and dermatitis activity in swiss albino mice. In addition, a significant inhibitory effect on blood histamine levels was observed. Mast cell membrane stabilization, inhibiting NO and histamine pathway was the proposed mechanism responsible of this activity [50].  

Conclusion 

Aristolochia is an important genus, in terms of its medicinal value, in the family Aristolochiaceae. As summarized in the present review, we may conclude that plants of the genus represent a promising source of new drugs for treating various diseases. 


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Alviano WS, Alviano DS, Diniz CG, Antoniolli AR., Alviano CS., Farias LM., Carvalho MAR., Souza MMG., Bolognese AM., 2008. In vitro antioxidant potential of medicinal plant extracts and their activities against oral bacteria based on Brazilian folk medicine. Archives of Oral Biology. 53, 545 � 552.

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Chitme HR, Malipatil M, Chandrashekhar VM, Prashant PM (2010) Antiallergic activity of Aristolochia bracteolata lank in animal model. Indian Journal of Experimental Biology. 48: 46 � 52. 
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