An overview on traditional uses and pharmacological profile of acorus calamus Linn. (Sweet flag) and other acorus species.
Acorns calamus (Sweet flag) has a long history of use and has numerous traditional and ethnomedicinal applications. Since ancient times, it has been used in various systems of medicines such as Ayurveda, Unani, Siddha, Chinese medicine, etc. for the treatment of various aliments like nervous disorders, appetite loss, bronchitis, chest pain, colic, cramps, diarrhea, digestive disorders, flatulence, gas, indigestion, rheumatism, sedative, cough, fever, bronchitis, inflammation, depression, tumors, hemorrhoids, skin diseases, numbness, general debility and vascular disorders. Various therapeutic potentials of this plant have been attributed to its rhizome. A number of active constituents from leaves, rhizomes and essential oils of A. calamus have been isolated and characterized. Of the constituents, alpha and betaasarone are the predominant bioactive components. Various pharmacological activities of A. calamus rhizome such as sedative, CNS depressant, anticonvulsant, antispasmodic, cardiovascular, hypolipidemic, immunosuppressive, anti-inflammatory, cryoprotective, antioxidant, antidiarrheal, antimicrobial, anticancer and antidiabetic has been reported. Genotoxicity and mutagenecity of beta and alpha-asarone is reported, which limits their use at high dosage. Though A. calamus has been used since ancient times, many of its uses are yet to be scientifically validated. In the present review an attempt has been made to explore traditional uses and pharmacological properties of A. calamus.
Contents Introduction Habitat and habit Common names Traditional uses Uses in traditional medicine Phytochemical studies Pharmacological and bioactivity studies Anti-inflammatory and immunomodulatory activity Antioxidant and protective effects Anticonvulsant and antispasmodic activity Actions on cardiovascular system (CVS) Actions on respiratory system Actions on nervous system Anti-diabetic properties Hypolipidemic properties Anticancer properties Antimicrobial properties Pesticidal properties Other bioactivities Toxicity studies Acknowledgements References
Acorus calamus Linn., commonly known as 'Sweet flag', is an aromatic medicinal plant, well known for its medicinal values. It is an integral part of the traditional Indian and Chinese systems of medicine and has a long history of use (Wu et al. 2009; Lee et al. 2011). In the vedic periods it was used as a 'rejuvenator' of the brain and nervous system. Charaka categorized it as a lekhaniya (natural substances that remove fat from the body), trptighna (anti-saturative), asthapanopaga (an adjunct to decoction enemas), sitaprasamana (relieves cold sensation on the skin), samjnasthapana (restores consciousness), vayasthapana (promotes longevity), arsoghna (anti-hemorrhoidal) and sirovirecana (cleansing nasal therapy) (Sharma 2000). It is widely used in traditional folk medicine of America and Indonesia for gastrointestinal disorders such as, colic pain, diarrhea and in the therapy of diabetes (Gilani et al. 2006; Si et al. 2010). It is also used in the treatment of cough, fever, bronchitis, inflammation, depression, tumors, hemorrhoids, skin diseases, numbness, general debility, as antidotes for several poisoning (Nadkarni 1998; Vaidyaratnam 1994). This plant is also described in Siddha medicine for its pediatric uses. Paste of the rhizome is used in rural areas of southern India to improve the speech and to memory in children (Meena et al. 2010).
Habitat and habit
A. calamus is a native of central Asia and eastern Europe, and is indigenous to the marshes of the mountains of India (Gupta 1964). It is cultivated throughout India, ascending to an altitude of about 2200 m. It is found/cultivated in the states of Jammu Kashmir, Himachal Pradesh, Manipur, Naga land, Uttarakhand, Uttar Pradesh, Tamil Nadu, Andhra Pradesh, Maharashtra and Karnataka (Pawar et al. 2011; Malabadi et al. 2007; Rao and Sreeramulu 1985).
A. calamus is a semi-evergreen perennial hairless herb that can grow to two meters high. It has a creeping rhizome. It is extensively branched and up to three centimeter in diameter. The rhizome is pale yellow to pinkish-brown on the outside and whitish but sometimes slightly pinkish on the inside. The upper surface is marked with large V-shaped leaf-scars and longitudinally furrowed. The surface beneath has circular pitted scars of rootlets arranged in irregular lines. Rhizome sections exhibit a large stele separated by a yellowish line, the endodermis from a thick cortex; numerous small, oval, vascular bundles are scattered throughout the section. Leaves are bright green having sword-shaped, based equitant, thickened in middle and wavy margins. Flowers of A. calamus have both male and female organs (Hermaphrodite), pollinated by insects (Prajapati et al. 2003; Nadkarni 1998; Wallis 1997).
English-Sweet Flag.; Chinese-Shi chang pu; Arabic-vash, vaj; French-acore calame; German-Kalmus; Italian-calamo aromatic; Dutch-kalmoeswortel; Hindi-Bajai, Gora-bach, Vasa Bach; Marathi-Vekhand; Tamil-Vashambu; Telugu-Vadaja, Vasa; Kannada-Baje; Malayalam-Vayambu; Sanskrit- Bhutanashini, Jatila. Vacha (Seidmann, 2005).
In Sanskrit, the language in which ayurveda is rendered, a number of synonyms are given to A. calamus (Table 1). The synonyms give clue about the properties of this drug.
The genus name, Acorus is derived from Acoron (coreon = the pupil of the eye) and the species calamus is derived from the Greek word, Calamos (a reed). The family Acoraceae comprises about 110 genera and more than 1800 species. The members of the family are rhizomatous or tuberous herbs. The genus Acorus comprises about 40 species, however, only few species like A. calamus (Linn.), A. christophii, A. tatarinowii (Schott.)and A. gramineus (Solandin Ait.) have been investigated for their chemical composition and bioactivities. A. calamus is extensively studied due to its medicinal and pharmacological significance (Ganjewala and Srivastava 2011).
Uses in traditional medicine
A. calamus rhizome has a long history of usage in many countries: at least 2000 years in China and India. Many native American tribes were familiar with calamus and it was used as an anesthetic for toothache and headaches. The ancient Chinese used it to lessen swelling and for constipation. The rhizome was also used by the ancient Greeks and included in the traditional remedies of many other European cultures.
A. calamus is used for the treatment of various ailments like appetite loss, bronchitis, chest pain, colic, cramps, diarrhea, digestive disorders, flatulence, gas, indigestion, nervous disorders, rheumatism, sedative, and vascular disorders (Kirtikar and Basu 1987). In the Ayurvedic system of medicine, the rhizomes of A. calamus are considered to possess aromatic, stimulant, bitter tonic, emetic, expectorant, emmenagogue, aphrodisiac, laxative, diuretic, antispasmodic, carminative, and anthelmintic properties. It is found to be effective in various disorders like chronic diarrhea, dysentery, bronchial catarrh, intermittent fevers, tympanitis, colic, otitis media, cough, asthma, and glandular and abdominal tumors (Anonymous 2001).
In Western herbal medicine the herb is chiefly employed for digestive problems such as gas, bloating, colic, and poor digestive function. Calamus helps distended and uncomfortable stomachs and headaches associated with weak digestion. Small amounts are thought to reduce stomach acidity, while larger doses increase deficient acid production. They are also employed for kidney and liver troubles, rheumatism, and eczema. In acidity, it is taken with honey and jaggary. In indigestion, vacha is taken with salt and water leads to emesis (Bangasen 1984). In Vamana therapy, it is used as emetic (Vantikrut) while in dyspepsia, it is employed as an appetizer (Vanhikrut) (Chunekar and Pandey 1998). It is widely used as a carminative (Vibandhanhara, Adhmanahara) in distension. It exerts antispasmodic (Shulaghni) effect by relieving abdominal pain. It removes stools (i.e. Shukrut Vishodhini) from body as well as improves its quality. It is also employed as mild diuretic (Mutravishodhini) which improves quality of urine. Vacha in combination with milk and water is useful in obstructive urinary disorders particularly in distended urinary bladder (Bangasen 1984). The decoction or powder of rhizome has been given in various pediatric aliments like cough, fever, abdominal pain, epilepsy etc. (Ignacimuthu et al. 2006; Chellaiah et al. 2006).
A. calamus rhizomes are used for the treatment of host diseases such as mental ailments like schizophrenia, psychoneurosis, insomnia, hysteria, epilepsy and loss of memory in the Indian ayurvedic system of medicine (Prajapati et al. 2003; Nadkarni 1998). Vacha taken with milk for one month improves 'pragnya' and 'shrutidharana' i.e. intellectual, grasping and memory (Shah 2005). Traditionally the new born child is given vasambu (rhizome paste) with honey and gold for proper brain development, speech ability, better visual power, increased seminal power. It stimulates nervous system (i.e. vatanadi sasthana) to get relief from depression. It is useful in improving speech in stammering and other disorders. After birth its paste is applied on tongue with ghee, gold and water to improve memory and grasping qualities (Jadhav 1994). It is a good sedative so that the extract is used for epilepsy, insanity and as a tranquillizer along with Valeriana jatamansi and Nardostacysgrandiflora. It is an ingredient of the ayurvedic preparation "Brahmi Bati" (Budhivardhar) which is indicated in epilepsy, coma, and hysteria and in cases of mental retardation; the same uses are prescribed for an Acorus containing Unani drug "Ma'jun Baladur". In headache, drowsiness, sinusitis, more sleepy feelings the 'Pradhaman nasya' i.e. its dry powder is inhaled in nostrils (Jadhav 1994).
A. calamus rhizomes are found to be very useful as a topical agent in skin related problems. The rhizomes are used in the form of powder, balms, enemas, and pills and also in ghee preparations (Kirtikar and Basu 1987; Anonymous 2001). A tub bath in the decoction of vacha, kustha (Savccera lappa) and vidanga (Embelina ribes) is useful in eczema and other skin diseases. It acts as 'sagnyasthapaka' i.e. it restores sensation useful in various comatose conditions. Its powder is sprinkled on infective and wounds with maggots. On baby's head it is applied in powder form after bath for protecting from cold. After bath its powder is applied to body like talcum powder (Kirtikar and Basu 1987).
The skin of the rhizomes is said to be haemostatic. In hemorrhoids, its fumes are given to pile masses to reduce swelling and pain. In the treatment of hemorrhoids, non-bleeding hemorrhoids are fomented with Poltice (Lukeworm) of vacha and Antheum sowa (Shatapushpa). Its application with mustard seeds paste is useful in hydrocele. In earache and tinnitus, vacha swarasa (juice) is poured in ear. Vacha is prescribed as 'Vedanasthapalea' (analgesic) in arthritis, rheumatoid arthritis, inflammatory conditions as external application in the form of paste (Lepa) (Shastri and Pandey 1997).
In Ayurveda, A. calamus is described as 'Lekhana' (lipid lowering action) (Table 2). The decoction of Vacha and Nimba (Azadirecta indica) is given for emesis in cardiac disease. It has lipid scavenging property that removes excessive fats from the body. Sometimes it is also prescribed along with honey, hot water and ava (Barley) in cardiac diseases (Bangasen 1984).
A wide variety of chemical constituents have been reported from the rhizomes, leaves and essential oil of Acorus calamus (Namba 1993; Wang et al. 1998). The content and composition of chemical constituents in plant parts vary with geographical condition, plant age, climate and ploidy of the plant (Venakutonis and Dagilyte 2003). According to Ogra et al. (2009) and Zhang (2005) the different Acorus species appear to follow a geographical pattern of distribution with respect to ploidy level.
A total of fifty three organic volatile compounds of rhizomes of Nepalese Acorus calamus L were isolated and identified, which belongs to alcohol (11), aldehyde (14), ester (3), furan (1), hydrocarbon (19), ketone (4), N-containing miscellaneous (1). [beta]-Asarone (46.78%) was found to be a major bioactive compound (Gyawali and Kim 2009).
At least one hundred and eighty five compounds in the oil of the triploid European A. calamus var. calamus, and ninety-three compounds in the oil of the tetraploid Indian A. calamus var. angustatus with f-asarone as the major constituent is reported. Sixty-seven hydrocarbons, fifty three carbonyl compounds, fifty six alcohols, eight phenols, two furans and four oxido compounds were detected in an alcohol extract of A. calamus var. calamus (Motley et al., 1994).
The content of [beta]-asarone in essential oil of Acorus spp. varies with the grade of polyploidy of the various Acorus cytotypes, sub varieties and/or species. [beta]-Asarone (90-96%) is abundantly found in tetraploid variety. In A. calamus var, angustata ENGER (tetraploid), about 80% [beta]-asarone is reported. In triploid plants (e.g. A. calamus var. calamus L.), 5% [beta]-asarone was present in the oil, while diploid plants such as A. calamus var. americanus WULFF lacked [beta]-asarone, but it has high amount of geranyl acetate (Wagner et al. 2011). In the rhizomes of some Chinese Acorus tatarinowii samples, formerly known as Acorus gramineus, an uncommonly high amount of [beta]-asarone (up to 80%) were detected (Wagner and Urlich-Merzenich 2013). The percentage of chemical components varies depending on the part of the plant from which the oil is extracted (Motley 1994). [beta]-Asarone [(Z)-asarone] is the major constituent in the leaves (27.4-45.5%), whereas acorenone is dominant in the rhizomes (20.86%) followed by isocalamendiol (12.75%). Monoterpene hydrocarbons, sequestrine ketones, (trans- or Alpha) Asarone (2,4,5-trimethoxy-l-propenylbenzene), and beta-asarone (cis-isomer) and eugenol were also identified (Balakumbahan et al. 2010; Raja et al. 2009). Other constituents such as alkaloids, flavanoids, gums, lectins mucilage, phenols, quinine, saponins, sugars, tannins and triterpenes are also recorded from this plant. Various sugars such as maltose (0.2%), glucose (20.7%) and fructose (79.1%) are reported (Balakumbahan et al. 2010). Calamenone (a tricyclic sesquiterpene) as well as calamendiol and isocalamendiol (both sesquiterpenes) occur in the roots. The volatile oil also contains terpenoids like calamine, calamenol, calamenone, eugenol, camphene, pinene and asaronaldehyde. Acorafuran is a sesquiterpenoid found in Calamus oil (Pandy et al. 2009).
[beta]-Asarone, geranylacetate, methyleugenol, cismethylisoeugenol, [beta]-farnesene, shyobunone, epishyobunone and isoshyobunone are abundantly present in the essential oil. The other chemical components include [alpha]- and [gamma]-asarone, calamenene, asaronaldehyde, acorenone, calamenone, n-heptanic acid, calamendiol, numerous sesquiterpenes, tannins, starches, mucin, soft gums and resins (Motley et al., 1994).
Other compounds identified in A. calamus were 4-terpineol, 2-allyl-5-ethoxy-4-methoxyphenol, epieudesmin, lysidine, spathulenol, borneol, furylethyl ketone, nonanoic acid, 2,2,5,5tetramethyl-3-hexanol, bornyl acetate, galgravin, retusin, (9E,12E,15E)-9,12,15-octadecatrien-l-ol, butyl butanoate, geranyl acetate, sakuranin, acetic acid, camphor, isoelemicin, a-ursolic acid, acetophenone, dehydroabietic acid, isoeugenol methylether, apigenin 4',7-dimethyl ether, dehydrodiisoeugenol, linalool, elemicin, linolenic acid (Balakumbahan et al. 2010).
Pharmacological and bioactivity studies
Rhizomes and its essential oils possess important bioactivities such as immunomodulatory and anticellular (Jayaraman et al. 2010; Mehrotra et al. 2003; Kim et al. 2009; Lad et al. 2010), antidiabetic (Wu et al. 2009; Lee et al. 2011; Lee et al. 2010; Si et al. 2010), antitumor/anticancer (Gaidhani et al. 2009; Chaitali et al. 2010) and antimicrobial properties (MacGaw et al. 2002; Phongpaichit et al. 2005; Devi and Ganjewala 2009).
Anti-inflammatory and immunomodulatory activity
Several studies have recognized the anti-inflammatory potential of A. calamus. The anti-inflammatory activity of A. calamus in rats using acute and chronic experimental models is evaluated. The oral administration of the extract showed inhibition of the carragenin-induced paw edema, inhibition of cotton pellet granuloma formation, and inhibition of croton oil granuloma pouch inflammatory response. The rhizomes extract showed significant anti-inflammatory effect in acute, chronic, and immunologic models of inflammation (Varde et al. 1988; Vohra et al. 1989). In another study anti-inflammatory activity of A. calamus leaf extract have been elucidated using human keratinocyte HaCaT cells. The extract inhibited production of pro-inflammatory cytokines through multiple mechanisms (Kim et al. 2009). Acetone extract of A. calamus displayed anti-inflammatory response in albino rat, where, inflammatory effect was completely diminished and the normal status of paw was achieved when 25-75% acetone extract was tested against inflammation within 30 min (Lad et al. 2010).
Mehrotra et al. (2003) demonstrated immunomodulatory properties of ethanolic extract of A. calamus rhizome. The extract inhibited proliferation of mitogen (phytohaemagglutinin; PHA) and antigen (purified protein derivative; PPD)-stimulated human peripheral blood mononuclear cells (PBMCs), nitric oxide and interleukins-2 production.
Antioxidant and protective effects
Several researchers have evaluated antioxidant potential of Acorns spp. and validated its protective roles in free radical and reactive oxygen species (ROS) generated disorders. The ethyl acetate extract of A. calamus exhibited strong antioxidant effect by inhibiting l,l-diphenyl-2-picrylhydrazyl (DPPH) free radical (Acuna et al. 2002). In another in vitro experiment, maximum DPPH scavenging activity of 86.43% was recorded at 0.2g/ml of extract (Govindarajan et al. 2003). The free radical scavenging activity of A. calamus has been found to be useful to overcome excess production of ROS generated due to continuous exposure to loud noise (Manikandan and Devi 2005). The ethyl acetate and methanolic extracts of A. calamus have protected most of the changes induced by noise stress in the rat brain. These changes were evaluated by measurement of the activities of superoxide dismutase, catalase, glutathione peroxidase, levels of reduced glutathione, level of vitamin C, E, protein thiols and lipid peroxidation. P-Asarone is believed to be involved in reducing the stress (Manikandan and Devi 2005). In another study, the antioxidant property of a-asarone was demonstrated against noise-stress-induced changes in the rat brain. In this study, a-asarone was administered intra-peritoneally one-half hour before the animals were exposed to noise-stress for 30 days. The antioxidant activity was measured by assessing the activity of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), levels of reduced glutathione (GSH), vitamin C, vitamin E, protein thiols and lipid peroxidation (LPO) in different regions of the rat brain. [alpha]-Asarone exhibited protective effect by normalizing the increased SOD and LPO, and decreasing CAT, GPx, GSH, vitamins C and E, and protein thiols (Manikandan and Devi 2005).
Palani et al. (2010) studied the antioxidant activities of ethanolic extract of A. calamus on acetaminophen induced toxicity in male albino rats. Acetaminophen increased the level of hemoglobin, total leukocyte count, packed cell volume, differential leukocyte count (DLC), mean corpuscular volume, granulocytes, raised body weight, uric acid and platelet concentration. A. calamus extract significantly increased activities of the renal superoxide dismutase, glutathione peroxidase, catalase and decreased the level of monodialdehyde content of acetaminophen-treated rats. A. calamus extract inhibited hemolysis caused by acetaminophen. Histopathological studies showed the protective nature of the ethanolic extract against acetaminophen induced necrosis and renal damage in rats (Palani et al. 2010).
A. calamus rhizome and leaves are traditionally used for the treatment of various neurological disorders due to its neuroprotective properties. Hydro-alcoholic extract of A. calamus has demonstrated neuroprotective effects in the middle cerebral artery occlusion-induced ischemia in rats (Shukla et al. 2006). [alpha] and [beta]-Asarone inhibited N-Methyl-D-Aspartate (NMDA) and glutamate induced excitotoxicity in primary cortical cultures (Cho et al. 2002).
Studies by Sandeep and Nair (2010) have demonstrated in vitro radio-protective effects of A. calamus. The protective effects were evaluated by measuring the degree of lipid peroxidation. In vitro DNA damage was measured by assessing the radiation induced relaxation of supercoiled plasmid DNA (pBR322) and alkaline single cell gel electrophoresis or comet assay was used to analyze any damage to cellular DNA. The free radical scavenging property of A. calamus safeguarded DNA and membrane damage caused due to [gamma]-radiations in murine cells and human peripheral blood leukocytes.
Anticonvulsant and antispasmodic activity
The anticonvulsant effect against the pain models in mice was observed when methanol extract of A. calamus was administered orally at the doses of the 100 and 200mg/kg. The anticonvulsant effect was studied through the Pentylenetetrazol-induced seizures method. This study suggested that the anticonvulsant effects might be potentiated by the activity of gamma aminobutyric acid (GABA) (Jayaraman et al. 2010). Calamus oil has also been reported for its antiepileptic property in adult albino mice, where it successfully prevented seizures in maximal electroshock seizure test (Khare and Sharma 1982). [alpha]-Asarone has shown a tendency to protect against metrazole mediated convulsions (Sharma et al. 1961).
In another study, A. calamus crude extract was found to display anti-spasmodic activity. In the isolated rabbit jejunum preparation, the crude extract of A. calamus caused inhibition of spontaneous and high [K.sup.*] (80 mM)-induced contractions, with [EC.sub.50] values of 0.42 and 0.13 mg/ml respectively, resulting in spasmolytic activity which is mediated through the calcium channel blockade (CCB). Results of the study suggest that the spasmolytic effect of the plant extract is mediated through the presence of CCB-like constituent(s), which is concentrated in the n-hexane fraction, and this study provides a strong mechanistic base for its traditional use in gastrointestinal disorders such as colic pain and diarrhea. Additionally, the methanolic extract of A. calamus exhibited anti-diarrheal potential against castor oil-induced diarrhea. The extract significantly reduced induction time of diarrhea and total weight of the feces (Shoba and Thomas 2001). The essential oil was found to be more antispasmodic than the alcohol and aqueous extracts (Bose et al. 1960). Further, it was observed that a-asarone is more active than that of essential oil (Das et al. 1962).
Actions on cardiovascular system (CVS)
A. calamus extract has been reported for its properties of lowering blood pressure and vascular modulation (Shaha and Gilani 2010). The essential oil was found to combat auricular fibrillation, auricular flutter, and ventricular arrhythmias after two-stage coronary ligation in dogs. It prolonged the conduction time and refractory period in isolated rabbit auricles (Madan et al. 1960). The alcoholic extract of A. calamus exhibited a dose dependant hypotensive action on blood pressure of dog (Moholkar et al. 1975). In a clinical study on forty-five patients of ischemic heart disease, the health of A. calamus-treated group was found to be significantly improved. A. calamus was effective in the improvement of chest pain, dyspnea of effort, reduction of body weight index, improving ECG, decreasing serum cholesterol and serum low-density lipoprotein (SLDL), and increasing serum high-density lipoproteins (SHDL) (Mamgain and Singh 1994).
Actions on respiratory system
A. calamus has been found to be a famous remedy for the respiratory disorders due to the unique combination of airways relaxant constituents that were found in the crude extract of A. calamus such as papaverine-like dual inhibitor of calcium channels and phosphodiesterase in the hexane fraction and anticholinergic, rolipram-like phosphodiesterase-4 inhibitor in the ethyl acetate fraction. The associated cardiac depressant effect has provided a pharmacological basis for the traditional use of A. calamus in the treatment of the disorders of airways such as asthma (Jabbar and Hassan 2010; Shaha and Gilani 2010).
In a clinical trial of patients with moderate to severe bronchial asthma, the fresh rhizome of A. calamus was administered by a chewing method for 2-4 weeks. The A. calamus was found to have antiasthmatic potential without any side effects (Rajasekharan and Srivastava 1977). In another study, small pieces of the rhizome were administered to asthmatic patients by a chewing method, and significant effect in relieving of bronchospasm was observed without any side effects (Chandra 1980).
Actions on nervous system
The in vitro acetyl cholinesterase (AChE) inhibitory potential of the hydroalcoholic extract and of the essential oil from A. calamus rhizomes and that of its major constituents was evaluated based on Ellman's method in 96-well microplates using bovine erythrocytes. The [IC.sub.50] values obtained for the hydroalcoholic extract, the essential oil, beta-asarone and alpha-asarone were 182.31 [+ or -] 16.78 [micro]g/ml, 10.67[+ or -] 0.81 [micro]g/ml, 3.33[+ or -] 0.02 [micro]M and 46.38 [+ or -] 2.69 [micro]M, respectively. The A. calamus essential oil and its constituents exhibited significant AChE inhibitory potential. [beta]-Asarone showed the maximum inhibitory potential (Mukherjee et al. 2007). Methanolic extract of A. calamus displayed significant AChE inhibition at a concentration 200 [micro]g/ml (Oh et al. 2004). Because cognitive performance and memory are related to acetyl choline levels, the AChE-inhibitory effect of the plant may account for its traditional use.
A. calamus is well-known traditionally for its sedative properties. The volatile fraction of the petroleum ether extract potentiated the sedative activity with pentobarbital, hexobarbital, and ethanol in mice (Dandiya and Cullumbine 1959). The essential oil showed sedative-tranquilizing action in rats, mice, dogs, and enhanced motor activities in mice. It was observed that a high dose of oil inhibited monoamine oxidase activity (Dhalla and Bhattacharya 1968).
A. calamus extract has the potential to be used in the treatment of diabetes (Wu et al. 2009). Ethanol extract of A. calamus has been reported to enhance differentiation in adipocytes which is very useful in the treatment of type 2 diabetes. However, [beta]-asarone from essential oil of A. calamus has shown inhibitory effect on adipogenesis in 3T3-L1 cells. It has been suggested that [beta]-asarone might have suppressed the expression of adipogenic transcription factors (Lee et al. 2011). In earlier study, the same group has reported that asarones inhibit adipogenesis and may reduce intracellular triglyceride levels by stimulating the phosphorylation of hormone sensitive lipase which triggers Iipolysis in 3T3-L1 adipocytes (Lee et al. 2010). A. calamus extract has also been reported to cause suppression of blood glucose level in the normal mice. A. calamus extract had hypoglycemic effects and alpha glucosidase inhibition and improved the postprandial hyperglycemia and cardiovascular complications (Si et al. 2010).
A. calamus extract demonstrated its cholesterol-reducing effect by decreasing cholesterol biosynthesis in the liver (D'Souza et al. 2007). The alcoholic extract of A. calamus containing saponins, prevent the cholesterol absorption and interferes with its enterohepatic circulation and also increase its fecal excretion (Parab and Mengi 2002).
The hypolipidemic mechanism of action of [alpha]-asarone has been established in a rat model where it is shown to inhibit hepatic HMG-CoA reductase (Rodriguez-Paez et al. 2003). In silico studies have revealed that [alpha]-asarone binds to the active site of HMG-CoA reductase. The methoxy groups play a key role in the binding and probably also in its biological activity, as shown by extensive SAR studies reported for analogs of [alpha]-asarone (Medina-Franco et al. 2005).
A. calamus rhizomes have been reported to have promising antiproliferative activities (Gaidhani et al. 2009; Chaitali et al. 2010). Lectins derived from A. calamus rhizomes shows potent antimitogenic activity toward mouse splenocytes and human lymphocytes. These lectins also significantly inhibited the growth of J774, a murine macrophage cancer cell line and, to a lesser extent, WEHI-279, a B-cell lymphoma cell line (Bains et al. 2005). The ethanolic extract of A calamus exhibited in vitro anticellular property by inhibiting production of nitric oxide, interleukin-2, and tumor necrosis factor-[alpha] (Mehrotra et al. 2003).
Epieudesmin and galgravin from methanolic extracts of A. calamus leaves have been identified as anticancer agents. Epieudesmin have antineoplastic activity against the murine P388 lymphocytic leukemia cell line and several human cancer cell lines (BXPC-3, MCF-7, SF268, NC1-H460, KM20L2, and DU-145). Galgravin prevented neuronal death and stimulating neurite growth. Studies have also suggested that the anticancer activity of calamus oil may be attributed to [beta]-asarone (Palani et al. 2010).
Phongpaichit et al. (2005) reports promising antifungal activity of A. calamus extracts against Trichophyton rubrum, Microsporum gypseum and Penicillium mameffei with [IC.sub.50] values of 0.2, 0.2 and 0.4mg/ml, respectively. However, it showed moderate activity against yeasts: Candida albicans, Cryptococcus neoformans and Saccharomyces cerevisiae (MIC 0.1-1 mg/ml). Scanning electron microscopic studies revealed that hyphae and conidia treated with extract were shrunken and collapsed due to cell fluid leakage (Phongpaichit et al. 2005). The minimum inhibitory concentration (MIC) of the rhizome and leaf extracts for fungi, Aspergillus niger, A. flavus and Microsporum canis was achieved at 2-4 mg/ml whereas, against yeasts Cryptococcus gastricus and C. albicans it was relatively higher i.e. 4-8 mg/ml. In addition, authentic [alpha]- and [beta]-asarones were also tested for their antimicrobial potential. Both [alpha]- and [beta]-asarones exhibited very strong antimicrobial activities against the fungi and yeasts than those of rhizome and leaf extracts. The study clearly suggested that A. calamus rhizomes and leaves must possess active principle [alpha]- and [beta]-asarones which is believed to be responsible for their antimicrobial activities, further it was established that [beta]-asarone has high antimicrobial activity as compared to the [alpha]-asarone (Devi and Ganjewala 2009).
Rajput and Karuppayil (2013) demonstrated anticandida properties of A. calamus rhizome (ethyl acetate) extract and its active principle, [beta]-asarone. [beta]-Asarone exhibited promising growth inhibitory activity at 0.5 mg/ml and it was fungicidal at 8 mg/ml. Minimum fungicidal concentration (MFC) of [beta]-asarone was highly toxic to C. albicans, killing 99.9% inoculum within 120 min of exposure. [beta]-Asarone inhibited C. albicans morphogenesis and biofilm development at sub-inhibitory concentrations. [beta]-Asarone was non-toxic to human RBCs, even at concentrations approaching minimum inhibitory concentration (MIC) value. Dose dependant reduction in ergosterol content was observed in [beta]-asarone treated cells where complete inhibition was achieved at growth inhibitory concentration, indicating the growth inhibitory effect of [beta]-asarone through inhibition of ergosterol biosynthesis (Rajput and Karuppayil 2013). A. calamus leaves extract show peroxidase activity. The enzyme was purified and evaluated through the chromatography and peak giving fractions were tested for the antifungal activity by gel filtration using Superose 1210/300 GL column (Ghosh 2006).
The leaf and rhizome part of A. calamus is found to possess the antibacterial activity. A. calamus rhizomes exhibit strong antibacterial activity against P. aeruginosa, S. aureus, B. subtilis showing MIC at 0.25 (Sabitha et al. 2003). Mycobacterium spp. and B. subtilis were susceptible to calamus oil (Radusiene et al. 2006). Devi and Ganjewala (2009), demonstrated that the rhizome and leaf ethyl acetate extract did not inhibit Gram +ve and -ve bacteria except E. coli strains. Similar activity was observed with authentic [alpha] and [beta]-asarone (Devi and Ganjewala 2009). Various studies suggest that the antibacterial activity may be attributed to a and [beta]-asarone (Bhuvaneswari and Balasundaram 2006; Devi and Ganjewala 2009).
Badam (1995) reported that the alcohol extract of the rhizome showed potent antiviral activity against Herpes Simplex Virus HSV-1 and HSV-2 at a concentration well below the cytotoxic concentration. Pretreatment of vero cells with the extract did not inhibit viral replication of HSV-1 and HSV-2. It shows that host cells were not affected by the extract. [beta]-asarone possesses strong inhibitory activity against the replication of both virus types. The crude alcohol extract and b-asarone showed toxicity to the host cells (Badam 1995).
The essential oils of the Pakistanian A. calamus exhibit potential pesticidal activity and also found to be effective on the cuts and wounds (Tariq et al. 2010). Essential oil was toxic against late 3rd instar larvae of Dengue fever virus vector mosquito, the Aedes aegypti. The [LC.sub.50] was found to be 1250 (Tariq et al. 2009). Asarones (2,4,5-trimethoxypropenyl-benzenes) isolated from the essential oil of A. calamus L. rhizomes, exhibited growth inhibitory and anti-feedant effect to the variegated cutworm (Balakumbahan et al. 2010). The repellent effect of petroleum ether extract of A. calamus has been investigated against Tribolium castaneum. 2,4,5-Trimethoxybenzaldehyde was identified as bioactive compound by [sup.1]H NMR, [sup.13]C NMR, H-H Cozy and HMBC spectra analyses (Hossain et al. 2008).
The pharmacological properties of A. calamus and its active constituents are shown in their entirety in Table 3.
A. calamus extract prevented the development the Fe[Cl.sub.3] induced epileptogenesis by modulating antioxidant enzymes (Pradhan et al. 2007). Anti-mutagenic effect of A. calamus has been studied using Salmonella typhimurium tester strains, where it showed decrease in revertants colonies against Na[N.sub.3] induced mutagenecity (Aqil et al. 2008).
It is demonstrated that [beta]-asarone is potentially toxic and carcinogenic (Keller and Stahl 1983; Taylor et al. 1967). In this study, rats were fed with diets containing various concentrations of [beta]-asarone for two years. The tumors were identified as leiomyosarcomas of the small intestine and were found in 800 ppm (0.08%) and 2000 ppm (0.2%). Also thrombosis within the chambers of the heart was observed in the 800 and 2000 ppm (0.08 and 0.27%) (Taylor et al. 1967). [beta]-asarone has an oral [LD.sub.50] of 1010 mg/kg bw in rats and an i.p. [LD.sub.50] of 184 mg/kg bw in mice (JECFA 1981). The oral [LD.sub.50] of calamus oil in rats is reported to be 8880 mg/kg bw (Opdyke 1977). Jenner et al. (1964) reported an oral [LD.sub.50] in rats of 777 mg/kg bw for Jammu calamus oil (containing approximately 75% [beta]-asarone).
It was found that [beta]-asarone at a dose of 5000 ppm (0.5%) showed mutagenicity in Salmonella typhimurium while [alpha]-asarone was inactive at 5000 ppm (Hsia et al. 1979). However, there are no reports on in vivo genotoxicity of [beta]-asarone. A study by Chamarro et al. (1998) demonstrated that [alpha]-asarone has hepatocarcinogenic and mutagenic activity in mice. In this study a dominant lethal mutation as well as direct [alpha]-asarone toxicity to spermatozoa has been observed.
In acute and chronic toxicity experiments, ethanolic extract of A. calamus did not cause significant changes in Winstar rats. This study concluded that the ethanolic extract of A. calamus does not have toxicity on acute and chronic administration in Winstar rats (Shah et al. 2012). There was no significant toxicity in rodents when orally administered with hydroalcholic extract of A calamus (Muthuraman and Singh 2012). Moreover, the rhizome (but not the isolated essential oil) has been used in India for thousands of years without reports of cancer which suggests that using the whole herb may be safe, though more research is needed (Chevallier 1996).
Received 10 June 2013
Received in revised form 19 August 2013
Accepted 29 September 2013
SBR is thankful to DST, New Delhi, for providing DST-1NSPIRE fellowship, Ref No. DST/INSPIRE FELLOWSH1P/2010/(290). We are thankful to Prof. S.B. Nimse, Hon'ble Vice Chancellor, SRTM University, for his kind support.
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Sandeep B. Rajput (a), Madan B. Tonge (b), S. Mohan Karuppayil (a), *
(a) DST-FIST and UGC-SAP Sponsored School of Life Sciences, SRTM University, Nanded 431-606, MS, India
(b) Prabhu Ayurvedic Clinic, Nanded 431-606, MS, India
* Corresponding author. Tel.: +91 9028528438: fax: +91 2462 229245.
E-mail address: email@example.com (S.M. Karuppayil).
Table 1 Synonyms of A calamus in Ayurveda. Sanskrit name Meaning Ugragandha Aggressive odor Shadgrandha Having multiple nodes Golomi Having hairs (as on a cow) Shatvarvika Having six nodes Lomasha Having hairs on nodes Aruna Reddish brown rhizomes Ikshuparni Leaves resembles sugarcane leaves Jatila Hairy rhizome Taxonomy (Seidemann, 2005) Kingdom: Plantae Subkingdom: Tracheobionta (Vascular plant) Superdivision: Spermatophyta (Seed plants) Division: Magnoliophyta (Flowering plants) Class: Liliopsida (Monocotyledons) Subclass: Arecidae Order: Arales Family: Acoraceae Genus: Acorus L. Species: calamus Synonyms: Acorus asiaticus Nakai; Acorus terrestris Spreng. Table 2 Ayurvedic terms indicating properties of Acorus calamus. Ayurvedic term Use/properties Vantikrut Induces vomiting in Vamana therapy (a therapy where the patient is made to vomit) Vanhikrut Used as appetizer in dyspepsia Vibandhhara or Adhanahara Carminative Shulaghni Antispasmodic (relieves abdominal pain) Shakrut vishodhini Removes stool from body Mathrushodihni Act as a diuretic Bhodhaneeya Arousing consciousness Karshini Reduces body weight Rokshoghni Checks or destroys the organisms Bhutaharet/Jantuharet Antimicrobial or antihelminthic properties Anilhara or Vatanasaka Anti-inflammatory, analgesic, pain reducing Vednasthapaka Analgesic, anti-inflammatory, arthritis Lekhana Lipid lowering Swaralu Improving speech or voice Smarani Memory promoter Shleshmaghni Pacifies kapha Vijaya Victory over diseases Mangalya Helps to keep healthy Table 3 Pharmacological properties of Acorous calamus and its active constituents. Activity Active compound or extract Antifungal Candida albicans, Cryptococcus Essential oil, [beta]-asarone neoformans Epidermophyton floccosum, Microsporum gypseum Trichophyton mentagrophytes and T. rubrum Ascosphaera apis Essential oil Aspergillus oryzae, A. Essential oil nidulans, A. fumigates, Penicillum aculactum, Phomopsis destuctum Curvularia lunata, Altemaria Essential oil altemata Macrophomina phaseolina Essential oil Fusarium moniliforme, Trichosporium Essential oil vesiculosum Helminthosporium oryzae Essential oil Antibacterial Aeromonas hydrophila Essential oil, [alpha]-asarone, [beta]-asarone Bacillus cereus, B. subtilis, Essential oil Shigella dysenteriae, Shigella flexneri, Vibrio cholera. Salmonella paratyphi. Pseudomonas pseudoalcaligenes. B. proteus. Aerobic spore Essential oil bearers, Staphylococcus pyogens. Shigella shiga Staphylococcus aureus, Essential oil Escherichia coli. Pseudomonas aeruginosa, Klebsiella pneumoniae Anti-inflammatory/immunomodulatory Anti-inflammatory effect in Leaf extract human HaCaT cells immunomodulatory activity in Ethanolic extract human PBMCs Anti-inflammatory activity in Acetone extract albino rats Anti-inflammatory activity in Rhizome extract rat models Anti-inflammatory activity in Leaf extract human keratinocytes Antioxidative/protective effect Antioxidant and Ethanolic extract nephroprotective effect in male albino rats Antioxidant activity in rats [alpha]-Asarone, ethyl brain acetate and methanolic extract Antioxidant activity Neuroprotective effect in Ethyl acetate extract ischemic rats Hydroalcoholic extract, [alpha]- and [beta]-asarone Anticonvulsant/antispasmodic Anticonvulsant activity in Methanolic extract, a-asarone mice models Antiepileptic activity in Calamus oil adult albino mice Antispasmolytic activity in Crude extract rabbit Prevents convulsions and Asarones electroshock seizures in rats Anticonvulsant action in Ethanolic extract amygdale kindled rats Anticancer Anti-carcinogenic activity in [alpha]-Asarone human carcinoma cells Anti-proliferative activity in Rhizome extract mice Anti-cancer activity against Epieudesmin and galgravin human cancer cells from methanolic extract Anticancer activity in human (3-Asarone from calamus oil cancer cells Antimitogenic activity in Lectins from rhizome mouse and human cancer lines Anticellular activity in human Ethanolic extract cancer cells Hypolipidemic Decreased cholesterol and Saponins from hydro-alcoholic triglyceride levels in rats extract Inhibited cholesterol Rhizome extract synthesis in rat liver Inhibited hepatic HMG-CoA [alpha]-Asarone reductase in rats Antidiabetic Exhibited antidiabetic effect Ethanolic extract by enhancing differentiation in adipocytes of mouse Suppress blood glucose levels Calamus extract in normal mice Cardiovascular related activity Lowers blood pressure in cats, Essential oil dogs and rabbits Hypotensive activity in dogs Alcoholic extract Cardiac depressant/antiasthmatic Airways relaxant activity Crude extract Antiasthmatic activity Rhizome extract CNS depressant/AChE-inhibitory Calming effect in monkeys Asarones AChE inhibitory activity in Hydroalcoholic and methanolic bovine erythrocytes extract, [alpha]-and [beta]-asarone Sedative effect in mice, dogs Volatile fraction of and rats petroleum ether extract, essential oil Hypnosis potentiating activity Volatile oil Activity Reference Antifungal Candida albicans, Cryptococcus Rajput and Karuppayil (2013), neoformans Epidermophyton Thirach et al. (2003) floccosum, Microsporum gypseum Trichophyton mentagrophytes and T. rubrum Ascosphaera apis Jatisatiener and Jatisatiener (1999) Aspergillus oryzae, A. Chantawannakul et al. (2005) nidulans, A. fumigates, Penicillum aculactum, Phomopsis destuctum Curvularia lunata, Altemaria Alankararao and Prasad (1981) altemata Macrophomina phaseolina Begum et al. (2004) Fusarium moniliforme, Trichosporium Ghosh (2006) vesiculosum Helminthosporium oryzae Saxena et al. (1990) Antibacterial Aeromonas hydrophila Bhuvaneswari and Balasundaram (2006) Bacillus cereus, B. subtilis, Chowdhury et al. (1993) Shigella dysenteriae, Shigella flexneri, Vibrio cholera. Salmonella paratyphi. Pseudomonas pseudoalcaligenes. B. proteus. Aerobic spore Alankararao and Prasad (1981) bearers, Staphylococcus pyogens. Shigella shiga Staphylococcus aureus, Chowdhury et al. (1993), Escherichia coli. Pseudomonas Parekh et al. (2006), aeruginosa, Klebsiella Rajendhran et al. (1998) pneumoniae Anti-inflammatory/immunomodulatory Anti-inflammatory effect in Kim et al. (2009) human HaCaT cells immunomodulatory activity in Mehrotra et al. (2003) human PBMCs Anti-inflammatory activity in Lad et al. (2010) albino rats Anti-inflammatory activity in Varde et al. (1988) rat models Anti-inflammatory activity in Kim et al. (2009) human keratinocytes Antioxidative/protective effect Antioxidant and Palani et al. (2010) nephroprotective effect in male albino rats Antioxidant activity in rats Manikandan and Devi (2005), brain Manikandan et al. (2005) Antioxidant activity Neuroprotective effect in Acuna et al. (2002) Shukla et ischemic rats al. (2006) Anticonvulsant/antispasmodic Anticonvulsant activity in Jayaraman et al. (2010), mice models Sharma et al. (1961) Antiepileptic activity in Khare and Sharma (1982) adult albino mice Antispasmolytic activity in Shoba and Thomas (2001) rabbit Prevents convulsions and Dandiya and Sharma (1962), electroshock seizures in rats Dandiya and Menon (1963) Anticonvulsant action in Hazra et al. (2005) amygdale kindled rats Anticancer Anti-carcinogenic activity in Hu and Ji (1986) human carcinoma cells Anti-proliferative activity in Gaidhani et al. (2009), mice Chaitali et al. (2010) Anti-cancer activity against Balakumbahan et al. (2010) human cancer cells Anticancer activity in human Palani et al. (2010) cancer cells Antimitogenic activity in Bains et al. (2005) mouse and human cancer lines Anticellular activity in human Mehrotra et al. (2003) cancer cells Hypolipidemic Decreased cholesterol and Parab and Mengi (2002) triglyceride levels in rats Inhibited cholesterol D'Souza et al. (2007) synthesis in rat liver Inhibited hepatic HMG-CoA Rodriguez-Paez et al. (2003) reductase in rats Antidiabetic Exhibited antidiabetic effect Wu et al. (2009) by enhancing differentiation in adipocytes of mouse Suppress blood glucose levels Si et al. (2010) in normal mice Cardiovascular related activity Lowers blood pressure in cats, Shaha and Gilani (2010), dogs and rabbits Dandiya and Cullumbine (1959) Hypotensive activity in dogs Moholkar et al. (1975) Cardiac depressant/antiasthmatic Airways relaxant activity Jabbar and Hassan (2010), Shaha and Gilani (2010) Antiasthmatic activity Rajasekharan and Srivastava (1977), Chandra (1980) CNS depressant/AChE-inhibitory Calming effect in monkeys Chak and Sharma (1965) AChE inhibitory activity in Mukherjee et al. (2007), Oh et bovine erythrocytes al. (2004) Sedative effect in mice, dogs Dandiya and Cullumbine (1959), and rats Dhalla and Bhattacharya (1968) Hypnosis potentiating activity Malhotra et al. (1962)
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|Author:||Rajput, Sandeep B.; Tonge, Madan B.; Karuppayil, S. Mohan|
|Publication:||Phytomedicine: International Journal of Phytotherapy & Phytopharmacology|
|Date:||Feb 15, 2014|
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