Antinociceptive, Antiinflammatory and Antibacterial Properties of Tamarix indica Roots
The present study investigated pharmacological activities to provide scientific basis to traditional usage of Tamarix indica. Phytochemical analysis of the dried roots of Tamarix indica (Tamaricaceae) indicated the presence of alkaloids, glycosides, flavonoids, saponins and tannins. The pharmacological interest of these compounds, coupled with the use of this plant in traditional medicine prompted the researchers to check for its possible antinociceptive, anti-inflammatory and antibacterial activities in animal models. The antinociceptive activity was studied using acetic acid-induced writhing in mice while anti-inflammatory tests were studied by using carrageenin induced rat paw edema model. Moreover, antibacterial activities were studied by using the disc diffusion method. The extract produced significant writhing inhibition in acetic acid-induced writhing in mice at the oral dose of 500 mg kg-1 body weight (p<0.001) comparable to the standard drug diclofenac sodium at the dose of 25 mg kg-1 of body weight. When given orally to rats at dose of 200 and 400 mg kg-1, the extract showed a significant (p<0.001) anti-inflammatory activity against carrageenin induced paw edema in rats comparable to the standard drug aspirin. Moreover, The extract of Tamarix indica exhibited significant in vitro antibacterial activity against Staphylococcus saprophyticus, Shigella sonnie, Salmonella typhi, Vibrio cholera, Streptococcus epidermidis, Shigella flexneri and Staphylococcus aureus with the zones of inhibition ranging from 10.76 to 16.34 mm. The obtained results provide a support for the use of this plant in traditional medicine and its further investigation.
January 24, 2011; Accepted: May 16, 2011;
Published: June 14, 2011
Tamarix indica (Family, Tamaricaceae), locally known in Bangladesh as
Nona jhau, is mainly growing up gregariously on newly formed alluvial
land rivers and by the coastal areas. These plants are mainly found as green,
branchlets shrub or small tree. It is distributed in the coast forests of Bengal,
Pakistan, Ceylon, Burma, Malay and Andamans. Different chemical constituents,
particularly from the roots, flower and bark, have been reported in the plant
(Ghani, 2003). This plant used for fuel wood and timber
in certain areas in the world (Khan et al., 2006a).
This plant is mainly found in the salty regions and is found between interdunal
areas of the desert (Qureshi and Bhatti, 2008). It can
be used as prophylactic and therapeutic remedies to cure malaria as folk medicine
(Tagarelli et al., 2010). The bark is bitter and
an astringent, tonic; fruit and roots are useful for dysentery and chronic diarrhoea
(Panhwar and Abro, 2007). The major chemical constituents
of Tamarix indica are tannin (50%), tamarixin, troupin, 4-methylcoumarin
and 3,3-di-O-methylellagic acid (Kirtikar and Basu, 1996;
Sehrawat and Sultana, 2006). Plants extract are rich
by acid compounds that are used as an inhibitor of nephrolithiasis (Bensatal
and Ouahrani, 2008). Several types of polyphenols (anthocyanins, tannins,
flavonones, isoflavonones, resveratrol and ellagic acid) are currently reported
(Sehrawat and Sultana, 2006). Ksouri
et al. (2009) also showed that the effects of antimicrobial activities
and presence of some antioxidant compound i.e. terpenoids (carotenoids and essential
oils). Presence of these compounds exhibit a wide spectrum of medicinal properties,
such as anti-allergic, anti-inflammatory, anti-thrombotic, cardio-protective
and vasodilatory effects (Balasundram et al., 2006;
Khan et al., 2006b). Literature reviews indicated
that no studies combining the antinociceptive, antidiarrhoeal and cytotoxic
activities of the roots have so far been undertaken. Taking this in view and
part of our ongoing search on Bangladeshi medicinal plants the present study
aimed at evaluating the antinociceptive, anti-inflammatory and antibacterial
properties of the roots extract of Tamarix indica.
MATERIALS AND METHODS
Plant material collection and extraction: The roots of Tamarix indica
were collected from the Sundarbans Mangrove Forests, Bangladesh in May
2009 and were taxonomically identified by experts at the Bangladesh National
Herbarium (accession number: 49566). About 400 g of powdered roots were taken
in a clean, flat-bottomed glass container and soaked in 1,300 mL of 80% methanol.
The container with its contents was sealed and kept for a period of 7 days accompanying
occasional shaking and stirring. The whole mixture then underwent a coarse filtration
by a piece of cotton followed by a filtration through Whatmann filter paper
grade 1 and the filtrate thus obtained was concentrated using a rotary evaporator
(Bibby RE200, Sterilin Ltd., U.K.) to get the crude extract.
Animals used: Young Swiss-albino mice of either sex, weighing 20-25
g, purchased from the Animal Research Branch of the International Centre for
Diarrhoeal Disease and Research, Bangladesh (ICDDR, B) were used for the test.
The animals were kept at animal house (Pharmacy Discipline, Khulna University)
for adaptation after their purchase under standard laboratory conditions (relative
humidity 55-65%, room temperature 25.0±2°C and 12 h light-dark cycle)
and fed with standard diets (ICDDR, B formulated) and had free access to tap
Drugs: Diclofenac (Square Pharmaceuticals Ltd, Bangladesh), Aspirin (Square Pharmaceuticals Ltd., Bangladesh), Gentamycin (Square Pharmaceuticals Ltd., Bangladesh).
Phytochemical tests: The crude extracts of Tamarix indica were
subjected to preliminary phytochemical screening for the detection of major
chemical groups. In each test 10% (w/v) solution of the extract in methanol
was used unless otherwise mentioned in individual test (Evans,
1989; Ghani, 2003).
Determination of antinociceptive activity: Antinociceptive activity
of the crude extract was tested using the model of acetic acid-induced writhing
in mice (Ahmed et al., 2007; Roome
et al., 2008). Analgesic activity of Tamarix indica extract
was compared to the inhibition of writhing of a standard analgesic agent (diclofenac
sodium). Experimental control mice were administered with 10 mL kg-1
1% Tween80 (Alpha labchem) with water; positive control mice were administered
with 25 mg kg-1 bodyweight diclofenac-sodium (Diclofenac) solution
made to 2.5 mL with water and two test concentrations (250 mg kg-1
and 500 mg kg-1 body weight) of the crude extract of Tamarix indica
was triturated by addition of small amount of Tween80 and water was slowly
added to make the final volume of the test solutions to 2.5 mL. Four groups
(Group1, 2, 3 and 4) of experimental animals were randomly selected with 5 animals
in each group for each treatment. Mice were carefully administered with Tween80,
diclofenac sodium and the test solutions by feeding needle. Thirty minutes interval
was given to ensure proper absorption. During the time test mice were noted
for any unwanted reactions. Acetic acid (0.7%) at a dose of 0.1 mg/10 g was
administered intraperitoneally to induce pain sensation. After an interval of
10 min which was given for the absorption of acetic acid, numbers of writhing
were calculated for 10 min.
Anti-inflammatory activity: Anti-inflammatory activity of Tamarix
indica was tested by using carrageenin induced rat paw edema model (Winter
et al., 1962; Ahmed et al., 2004).
Rats were randomly divided into four groups, each consisting of six animals.
Group I was kept as control giving 1% (v/v) tween-80 solution in
water; group II was kept as positive control and was given the standard
drug Aspirin at a dose of 150 mg kg-1 of body weight; group III and
IV were test groups, treated with extracts at the doses of 200 and 400 mg kg-1
of body weight respectively. Control vehicle, standard drug and the extracts
were given orally 1 h prior to the injection of 0.1 mL of 1% freshly prepared
suspension of carrageenin. The paw volume was measured by using a plethysmometer
(Ugo Basile 7140, Italy) just before and 1, 2, 3, 4 and 5 h after the carrageenin
Antibacterial activity: The antibacterial activity of Tamarix indica
extract was studied against Staphylococcus saprophyticus, Shigella
sonnie, Salmonella typhi, Vibrio cholera, Streptococcus epidermidis, Shigella
flexneri and Staphylococcus aureus clinical isolates. All bacterial
strains were kindly provided by IMTECH, Chandigarh (India). Cultures of these
bacteria were grown in a nutrient broth at 37°C and maintained on nutrient
agar (Himedia, India) slants at 40°C. The antibacterial property was studied
by the disc diffusion method (Chattopadhyay et al.,
2002) using extract 200 mg/disc. Control disks contained solvents only (50%
aqueous methanol).Gentamycin was used as positive controls. Minimum Inhibitory
Concentration (MIC) was evaluated by the micro dilution method using 5 mL of
liquid broth with different concentrations of extract (Ahmad
and Beg, 2001; Bayoud et al., 2007).
Statistical analysis: Students t-test was used to determine significant differences between the control group and test group.
|| Phytochemical properties of Tamarix indica crude roots
|+ve: Presence, -ve: Absence
|| Effects of Tamarix indica crude root extract on writhing
effect on acetic acid induced mice
|For test group the results were statistically significant
(p<0.001). -: Tamarix indica Crude Extract. 30 min after treatment,
0.7% acetic acid was injected i.p. 10 min after injection writhing responses
was recorded for 10 min. N = 5
|| Effect of methanolic extract of Tamarix indica on
carrageenin induced rat paw edema
|#Increase in paw edema volume (mL)x1000±SEM.
Values are expressed as Mean±SEM. (No. of animals, n = 6); *indicates
p<0.05, **indicates p<0.001 vs. control; Test group: Tamarix indica;
p.o.: Per oral
Phytochemical test: Phytochemical tests were performed with the crude extract of Tamarix indica and the results are tabulated in Table 1. The results manifest the presence of alkaloids, glycosides, flavonoids, saponins and tannins from crude extract of the roots.
Antinociceptive activity test: The methanolic extract showed significant inhibition of writhing when compared to the control (Table 2). At dose of 250 and 500 mg kg-1 of body weight, the extracts produced 47.94 and 64.47% inhibition in test animals, respectively. The results were found to be statistically significant (p<0.001) and were comparable to the standard drug diclofenac sodium, which showed about 85.95% writhing inhibition at dose of 25 mg kg-1 (p<0.001).
Carrageenin-induced rat paw edema: In the carrageenin induced rat paw
edema model of anti-inflammatory activity, the methanolic extract of roots of
Tamarix indica showed a significant inhibitory effect on the edema formation
from the first hour to fifth hour.
The highest inhibitory effect was found during the third hour where the inhibition
was 24.59% (p<0.001) and 40.28% (p<0.001) at the doses of 200 and 400
mg kg-1, respectively. These findings were comparable to standard
drug aspirin where the inhibition was 51.23% (Table 3).
In vitro antibacterial activity: Table 4 showed the extract of Tamarix indica exhibited significant in vitro antibacterial activity against Staphylococcus saprophyticus, Shigella sonnie, Salmonella typhi, Vibrio cholera, Streptococcus epidermidis, Shigella flexneri and Staphylococcus aureus with the zones of inhibition ranging from 10.76 to 16.34 mm.
Plants are employed as important source of medication in many traditional medications
(Grover et al., 2002; Keung
and Vallee, 1998; Neves et al., 2009). Since
Tamarix indica belongs to the coastal forests, part of the plant constituents
may be polar in nature. Methanol was used which has a wide range of solubility
in both polar and non-polar region. To avoid any solvent effect on the experimental
animals, the solvent was evaporated completely to dryness (Ahmed
et al., 2007).
Preliminary phytochemical screening of the extract showed the presence of alkaloids,
glycosides, flavonoids, saponins and tannins. Polyphenolic compounds, like flavonoids
and tannins, commonly present in mangrove plants have been reported to have
multiple pharmacological effect, including antinociceptive activity. Roome
et al. (2008) showed that plants containing flavonoids and pentacyclic
triterpenes may cause pain inhibition in mice. Presence of glycosides can cause
the antinociceptive activity.
Antonociceptive activity was explored with two different concentrations of
250 and 500 mg kg-1 body weight. Antinociceptive activity of Tamarix
indica was tested by acetic acid-induced writhing model in mice. Acetic
acid-induced writhing model causing pain sensation by tiggering localized inflammatory
response. Acetic acid, which is used to induce writhing, causes analgesia by
liberation of endogeneous substances, which in turn excite the pain nerve endings
(Taesotikul et al., 2003). Increased levels of
PGE2 and PGF2α in the peritoneal fluid have been
reported to be responsible for pain production caused by intraperitoneal administration
of acetic acid (Derardt et al., 1980). The result
of the test showed that the methanolic extract of the roots of Tamarix indica
at dose 500 mg per kg body weight exhibit significant writhing inhibition
(p<0.001) as compared with the standard drug diclofenac sodium (Table
2). The polar compounds present in the plant extract may be responsible
for the obtained antinociceptive activity. According to the basis of this result
it can be concluded that the extract possesses antinociceptive activity.
The most widely used primary test for the screening of new anti-inflammatory
agents is the carrageenin-induced rat paw edema model (Winter
et al., 1962). The edema formation is a biphasic event. The initial
phase, observed during the first hour, is attributed to the release of histamine
and serotonin (Vinegar et al., 1996) and the delayed
edema is due to the release of bradykinin and prostaglandins (Di
Rosa et al., 1971). It has been reported that the second phase of
edema is sensitive to steroidal and non-steroidal anti-inflammatory agents (Di
Rosa et al., 1971). The extract reduced the paw volume significantly
from 1 to 5 h in which the highest effects were found at the third hour. These
results tend to suggest the probable anti-inflammatory activity of the extract.
In this experiment, methanolic extract of Tamarix indica showed moderate sensitivity to the five of the test organisms both gram positive and gram negative type of bacteria. The highest zone on inhibition (16.34 mm) was recorded against Shigella sonnie. Moreover, the experiment was only conducted with five species of bacteria as test samples. Therefore further research is essential to evaluate the sensitivity of the plant extract against other species of bacteria, fungi, virus of other microorganisms.
Finally, it can be concluded from the study that the antinociceptive, cytotoxic and diuretic effects of the methanolic roots extract of Tamarix indica may be presence of different chemical compounds which works through the specific and non-specific mechanisms. However, extensive studies are needed to evaluate the precise mechanism(s), active principles and the safety profile of the plant as a remedy for different therapeutical conditions.
The authors are thankful to Prof. Dr. Samir Kumar Sadhu, Head, Pharmacy Discipline, Khulna University; Ahmed Ayedur Rahman, Assistant professor, Pharmacy Discipline, Khulna University; Dr. Mahiuddin Alamgir, Research Scientist, National Measurement institute (NMI), Australia for their encouragement during the research time. All the informants of the study area are cordially acknowledged for their valuable cooperation.
Ahmad, I. and A.Z. Beg, 2001. Antimicrobial and phytochemical studies on 45 Indian medicinal plants against multi-drug resistant human pathogens. J. Ethnopharmacol., 74: 113-123.
CrossRef | Direct Link |
Ahmed, F., A.H. Al Mamun, I.Z. Shahid, A.A. Rahman and S.K. Sadhu, 2007. Antinociceptive, antidiarrhoeal and cytotoxic activity of Aegiceras corniculatum. Int. J. Orient. Pharm. Exp. Med., 7: 191-196.
CrossRef | Direct Link |
Ahmed, F., M.S.T. Selim, A.K. Das and M.S.K. Choudhuri, 2004. Anti-inflammatory and antinociceptive activities of Lippia nodiflora Linn. Pharmazie, 59: 329-330.
Direct Link |
Balasundram, N., K. Sundram and S. Samman, 2006. Phenolic compounds in plants and agri-industrial by-products: Antioxidant activity, occurrence and potential uses. Food Chem., 99: 191-203.
CrossRef | Direct Link |
Bayoud, B., S.E. Djilani, B. Legseir, M.R. Ouahrani and A. Djilani, 2007. Antibacterial activity of ethanol extracts and total alkaloids of Datura Stramonium and Ruta Graveolens. J. Life Sci., 1: 78-81.
Bensatal, A. and M.R. Ouahrani, 2008. Inhibition of crystallization of calcium oxalate by the extraction of Tamarix gallica L. Urol. Res., 36: 283-287.
Chattopadhyay, D., G. Arunachalam, A.B. Mandal, T.K. Sur, S.C. Mandal and S.K. Bhattacharya, 2002. Antimicrobial and anti-infl ammatory activity of folklore: Mellotus peltatus leaf extract. J. Ethnopharmacol., 82: 229-237.
Deraedt, R., S. Jouquey, F. Delevallee and M. Flahaut, 1980. Release of prostaglandins E and F in an algogenic reaction and its inhibition. Eur. J. Pharmacol., 61: 17-24.
CrossRef | PubMed | Direct Link |
Di Rosa, M., P.J. Giroud and D.A. Willoughby, 1971. Studies on the mediators of the acute inflammatory response induced in rats in different sites by carrageenan and turpentine. J. Pathol., 101: 15-29.
PubMed | Direct Link |
Evans, W.C., 1989. Terace and Evan's Textbook of Pharmacognosy. 13th Edn., Cambridge University Press, London,.
Ghani, A., 2003. Medicinal Plants of Bangladesh-Chemical Constituents and Uses. 2nd Edn., The Asiatic Society of Bangladesh, Dhaka, Bangladesh, pp: 362-363, 502-505.
Grover, J.K., S. Yadav and V. Vats, 2002. Medicinal plants of India with anti-diabetic potential. J. Ethnopharmacol., 81: 81-100.
CrossRef | Direct Link |
Keung, W.M. and B.L. Vallee, 1998. Kudzu root: An ancient chinese source of modern antidipsotropic agents. Phytochemistry, 47: 499-506.
Khan, M.A., H. Barth, G.C. Kust and B. Boer, 2006. Sabkha Ecosystems: Volume II: The South and Central Asian Countries. Springer, Netherlands.
Khan, M.A., R. Ansari, B. Gul and M. Qadir, 2006. Crop diversification through halophyte production on salt-prone land resources. CAB Rev.: Perspect. Agric. Vet. Sci. Nutr. Natl. Resour., 1: 1-9.
CrossRef | Direct Link |
Kirtikar, K.R. and B.D. Basu, 1996. Indian Medicinal Plants. 2nd Ed., Indian Press, Allahabad, pp: 61-62.
Ksouri, R., H. Falleh, W. Megdiche, N. Trabelsi and B. Mhamdi et al., 2009. Antioxidant and antimicrobial acivities of the edible medicinal halophyte Tamarix gallica L. and related polyphenolic constituents. Food Chem. Toxicol., 47: 2083-2091.
Neves, J.M., C. Matos, C. Moutinho, G. Queiroz and L.R. Gomes, 2009. Ethnopharmacological notes about ancient uses of medicinal plants in Tras-os-Montes (Northern of Portugal). J. Ethnopharmacol., 124: 270-283.
Panhwar, A.Q. and H. Abro, 2007. Ethnobotanical studies of Mahal Kohistan (Khirthar National Park). Pak. J. Bot., 39: 2301-2315.
Direct Link |
Qureshi, R. and G.R. Bhatti, 2008. Diversity of micro-habitats and their plant resources in Nara desert. Pak. J. Bot., 40: 979-992.
Direct Link |
Roome, T., A. Dar, S. Ali, S. Naqvi and M.I. Choudhary, 2008. A study on antioxidant, free radical scavenging, anti-inflammatory and hepatoprotective actions of Aegiceras corniculatum (stem) extracts. J. Ethnopharmacol., 118: 514-521.
CrossRef | PubMed |
Sehrawat, A. and S. Sultana, 2006. Tamarix gallica ameliorates thioaceamide-induced hepatic oxidative stress and hyperproliferative response in Wistar rats. J. Enzyme Ihibit. Med. Chem., 21: 215-223.
Taesotikul, T., A. Panthong, D. Kanjanapothi, R. Verpoorte and J.J.C. Scheffer, 2003. Anti-inflammatory, antipyretic and antinociceptive activities of Tabernaemontana pandacaqui poir. J. Ethnopharmacol., 84: 31-35.
CrossRef | PubMed |
Tagarelli, G., A. Tagarelli and A. Piro, 2010. Folk medicine used to heal malaria in Calabria (southern Italy). J. Ethnobiol. Ethnomed., 6: 27-27.
Vinegar, R., W. Schreiber and R. Hugo, 1996. Biphasic development of carrageenin edema in rats. J. Pharmacol. Exp. Ther., 166: 96-103.
Winter, C.A., E.A. Risley and G.W. Nuss, 1962. Carrageenin-induced edema in hind paws of the rat as an assay for anti-inflammatory drugs. Proc. Soc. Exp. Biol. Med., 111: 544-547.
Direct Link |