|
|
|
|
Research Article
|
|
Antinociceptive Activity of Dalbergia spinosa Roxb. Stem Barks |
|
S.Z. Raihan,
M.M. Monir,
P. Biswas,
S.K. Biswas,
A. Chowdhury,
J. Das
and
A.C. Das
|
|
|
ABSTRACT
|
Dalbergia spinosa Roxb. (Family: Fabaceae) has many
traditional uses in Bangladesh. The crude methanol extract of the stem barks
of D. spinosa Roxb. was investigated for possible antinociceptive activity
using acetic acid induced writhing model in mice. Phytochemical analysis was
also performed using standard procedures to detect the presence of phytochemicals
in the crude plant extract. The study results showed 37.20% inhibition of writhings
in the tested mice when methanol extracts were given intraperitoneally (i.p.)
at the dose of 250 mg extract kg-1 b.wt. The maximum inhibition (40.01%)
of nociception effect was achieved at 500 mg extract kg-1 b.wt.,
i.p. which was also compared with the antinociceptive activity of the standard
drug, diclofenac sodium at the dose of 25 mg extract kg-1 b.wt. which
produced 68.37% inhibition of nociception effect. The inhibition of writhings
was calculated in respective to control group and it was found that p-values
(<0.0001) calculated by students t-test were statistically significant.
However, the phytochemical screening revealed the presence of alkaloid, steroid,
flavonoid, tannin, reducing sugar and gum. Finally, it can be concluded that
crude methanol extracts of D. spinosa stem barks contain biologically
active phytoconstituents exhibiting significant dose-dependent antinociceptive
activity in the mice model used. Thus, it is recommended to isolate and characterize
the compounds for the development of new analgesics.
|
|
|
|
|
Received: December 09, 2011;
Accepted: March 02, 2012;
Published: June 12, 2012
|
|
INTRODUCTION
Dalbergia spinosa Roxb. found in Bangladesh (Senthamarai
et al., 2003) was selected and tested to justify its medicinal use.
This medicinal plant belongs to the family of Fabaceae (Jena
et al., 2004). Traditionally, crude drugs have applications for the
treatment of a range of pain complications (Eidi et
al., 2011). D. spinosa is a mangrove plant in the Sundarban islands
and the medicinal plant is also found in India. It is a shrub having small white
flowers. Earlier researchers have identified the presence of glycosides in this
plant species (Anjaneyulu et al., 2005). Various
isoflavones have been isolated and characterized from the roots and stems of
D. spinosa (Kumar and Muller, 1999). The plant
also contains dalspinin, dalspinosin and dalspinin-7-O- β-D-galactopyranoside
(Senthamarai et al., 2003).
Moreover, the two new isoflavone galactosides such as prunetin 4'-O-β-d-galactoside
and 7-methyltectorigenin 4'-O-β-d-galactoside have been isolated from the
leaves and stem-bark of this medicinal plant (Narayanan
and Nacarajan, 1988). Fever, pain, skin infections and urinary tract infections
can be treated with D. spinosa (Senthamarai et
al., 2003). The roots of the plant possess significant anti-inflammatory
activity which is comparable to Indomethacin (Jaiganesh
and Senthamara, 2010) Different types of diseases are commonly treated by
the phytochemical compounds of the traditional medicinal plants (Pareta
et al., 2011). It is reported that the crude extracts have also significant
antimicrobial activities against both gram positive and gram negative bacteria
(Senthamarai et al., 2003). Due to potential
applications of D. spinosa in folkloric medicine, the present study was
conducted to investigate the antinociceptive activity of the crude methanol
extract of stem barks of the plant.
MATERIALS AND METHODS
Collection of plant materials: The stem barks of D. spinosa were
collected from Sundarban Islands on 2009 and subsequently identified by the
taxonomists of Bangladesh National Herbarium, Mirpur, Dhaka. The voucher specimen
of the plant was deposited at the Faculty of Pharmacy, University of Dhaka.
Preparation of plant extracts: The collected stem barks were separated from undesirable materials and they were air-dried for one week. The dried stem barks were ground into a coarse powder with the help of a suitable grinder. The powder was stored in an airtight container and kept in a cool, dark and dry place. About 350 g of powered material was taken in a clean, flat bottomed glass container and soaked in 900 mL of 80% methanol. The container with its contents was sealed and kept for a period of 10 days accompanying occasional shaking and stirring. The whole mixture then underwent a coarse filtration by a piece of clean and white cotton material. Then it was filtered through Whatman filter paper. The obtained filtrate was evaporated under ceiling fan and in a water-bath until dried. It rendered a gummy concentrate of reddish black color residue (yield 4.57%) which was designated as crude methanol extract of the stem barks of D. spinosa. Chemicals and standard drug: Acetic acid (0.7%) and methanol were collected from LOBA Chemicals Pvt. Ltd., India. And Diclofenac sodium was obtained from Square Pharmaceuticals Ltd., Bangladesh. All the chemicals used during the study were of analytical grade.
Preliminary phytochemical screening: Preliminary phytochemical analysis
of the methanol extracts was carried out using standard procedures (Trease
and Evans, 1989).
Animals and experimental model: The experiment of antinociceptive activity
was conducted on Swiss albino mice aged 4-5 weeks and weighing 25-30 g of both
sexes collected from the Animal Research Branch of the International Centre
for Diarrhoeal Disease and Research, Bangladesh (ICDDR, B). Before starting
of the experiment, all the mice were subjected to acclimatize for one week under
standard experimental conditions (relative humidity 40-65%, room temperature
25±2.0°C and 12 h light/12 h dark cycle). They were fed ICDDR, B
formulated rodent food and water ad libitum. The antinociceptive activity
was determined by acetic acid induced writhing method (Whittle,
1964). The Swiss albino mice were divided into four groups consisting of
5 mice per group.
The first group (Group I) served as control receiving 1% Tween solution in
water (10 mg kg-1 b.wt.), the second group (Group II: Positive control)
received Diclofenac Sodium (25 mg kg-1) orally 30 min before acetic
acid injection (0.7%, 1 mL 100 kg-1 i.p.) and third and fourth groups
received the extract at doses of 250 and 500 mg kg-1 i.p., respectively
before administration of acetic acid. The mice were then placed in individual
cages and the number of abdominal contractions was observed 5 min after stimulation
for a period of 10 min.
Statistical analysis: The percentage inhibition of writhing was obtained using the following formula:
The results of the experiment were expressed as means±Standard Error of Mean (SEM). Students t-test (GraphPad Software) was used to determine a significant difference between the control and experimental groups where p values of less than 5% (p<0.05) was chosen as the level of significance. RESULTS Preliminary phytochemical screening: The results of phytochemical analysis of the methanol extract of D. spinosa stem barks are summarized in Table 1. Phytochemical study revealed the presence of alkaloid, steroid, flavonoid, tannin, reducing sugar and gum. Antinociceptive activity: In acetic acid-induced writhing model, the crude methanol extract showed a significant dose-dependent decrease in the number of writhings. The extracts at the dose of 250 and 500 mg crude extract kg-1 body weight showed 37.20% and 40.01% inhibition of writhings in the experimental animals in a dose dependent manner. The obtained results were also comparable to the inhibition observed with a standard antinociceptive drug, Diclofenac sodium at 25 mg kg-1 b.wt. (77.21% inhibition). The results of antinociceptive activities are shown in Table 2.
DISCUSSION
Preliminary phytochemical analysis showed the presence of alkaloid, steroid,
flavonoid, tannin, reducing sugar and gum in the crude methanol extract of the
plant. It was revealed that the pain perception was inhibited by flavonoids,
alkaloids and tannins (Ramaswamy et al., 1985;
Zakaria et al., 2006; Rahman
et al., 2011) which were also confirmed by this study. Moreover,
flavonoids could show the anti-inflammatory, antioxidant and free radical scavenging
activities (Hossinzadeh et al., 2002; Okwu
and Orji, 2007). Thus, the plant might also possess anti-inflammatory and
antioxidant activities which have been investigating in our laboratory.
Table 1: |
Results of preliminary phytochemical screening of methanol
extract of D. spinosa (MEDS) stem bark |
|
+: Present, -: Absent |
Table 2: |
Effect of MEDS stem bark on acetic acid-induced writhing
in mice |
|
Values are expressed as Mean±SEM, n: No. of mice, CI:
Confidence interval |
Clinical tests have confirmed the efficacy and safety of traditional medicinal
plants to control pain and inflammation (Musa et al.,
2007; Narendhirakannan et al., 2007; Woode
et al., 2009). Acetic acid induced abdominal constriction is a sensitive
procedure to establish peripherally acting analgesics. The response is thought
to be mediated by the prostaglandin pathways (Le Bars et
al., 2001). The promising antinociceptive activity of the crude methanol
extract of D. spinosa stem barks might be due to the presence of analgesic
principles which interfere in the biosynthesis of prostaglandins and some other
autacoids. It has also been found that certain flavonoids play an important
role in the inhibition of production of prostaglandins which also inhibit the
key enzymes such as lipoxygenase, phospholipase and cyclooxygenase responsible
for prostaglandin biosynthesis. Inhibition of these enzymes provides the mechanism
by which flavonoids inhibit inflammatory processes (Manthey,
2000).
It is therefore possible that the inhibitory effects of nociceptive activities observed in the extract may be attributed in part to its flavonoid content. From the study, it may be concluded that the crude methanol extract of D. spinosa stem barks possesses significant antinociceptive activities. Thus, the results tend to corroborate the traditional use of this plant in the treatment of pain. However, further investigations are required to identify the active constituent(s) and to verify the therapeutic merits of the active constituent(s). CONCLUSION In conclusion, it can be claimed that D. spinosa possesses significant analgesic action which gives a scientific support to the traditional use of the plant in the management of pain. However, a lot of research work is required to identify the exact mode of action for antinociceptive activity and to isolate the active phytoconstituent(s) responsible for such potential. ACKNOWLEDGMENTS All authors are thankful to the authority of Square Pharmaceuticals Limited and International Centre for Diarrhoeal Disease and Research, Bangladesh for providing support to complete this research work successfully.
|
REFERENCES |
Anjaneyulu, A.S.R., V.L. Rao and K. Sreedhar, 2005. Chemical examination of the mangrove plant Dalbergia spinosa Roxb. Indian J. Chem., 44: 209-211.
Eidi, A., M. Eidi, V. Mozaffarian, A. Rustaiyan, A. Mazooji, Z. Khaboori and F. Nabiuni, 2011. Antinociceptive and anti-inflammatory effects of ethanolic extract of Salvia syriaca L. in mice. Int. J. Pharmacol., 7: 394-399. CrossRef | Direct Link |
Hossinzadeh, H, M. Ramezani, M. Fedishei and M. Mahmoudi, 2002. Antinociceptive, anti-inflammatory and acute toxicity effects of Zhumeria majdae extracts in mice and rats. Phytomedicine, 9: 135-141. PubMed |
Jaiganesh, K.P. and R. Senthamarai, 2010. Anti inflammatory activity of root extracts of Dalbergia spinosa Roxb. Adv. Pharmacol. Toxicol., 11: 33-36. Direct Link |
Jena, S., P. Sahoo, S. Mohanty and A.B. Das, 2004. Identification of RAPD markers, in situ DNA content and structural chromosomal diversity in some legumes of the mangrove flora of Orissa. Genetica, 122: 217-226. CrossRef | Direct Link |
Kumar, K.C.S. and K. Muller, 1999. Medicinal plants from Nepal: II. Evaluation as inhibitors of lipid peroxidation in biological membranes. J. Ethnopharmacol., 64: 135-139. CrossRef |
Le Bars, D.L., M. Gozariu and S.W. Cadden, 2001. Animal models of nociception. Pharmacol. Rev., 53: 597-652. PubMed | Direct Link |
Manthey, J.A., 2000. Biological properties of flavonoids pertaining to inflammation. Microcirculation, 7: S29-S34. Direct Link |
Musa, Y.M., A.K. Haruna, A.H. Yaro, A.A. Ahmadu, H. Usman, 2007. Analgesic and anti-inflammatory effects of leaf extracts of Pseudocedrella kotschyii Harms (Meliaceae). J. Pharmacol. Toxicol., 2: 542-550.
Narayanan, V. and N.S. Nacarajan, 1988. Two isoflavone galactosides from Dalbergia spinosa. Phytochemistry, 27: 2364-2365. Direct Link |
Narendhirakannan, R.T., S. Subramanian and M. Kandaswamy, 2007. Evaluation of anti-inflammatory activity of Cleome gynandra L. leaf extract on acute and chronic inflammatory arthritis studied in rats. J. Pharmacol. Toxicol., 2: 44-53. CrossRef | Direct Link |
Okwu, D.E. and B.O. Orji, 2007. Phytochemical composition and nutritional quality of Glycine max and Vigna unguiculata (L.) Walp. Am. J. Food Technol., 2: 512-520. CrossRef | Direct Link |
Pareta, S.K., K.C. Patra, P.M. Mazumder and D. Sasmal, 2011. Establishing the principle of herbal therapy for antiurolithiatic activity: A review. J. Pharmacol. Toxicol., 6: 321-332. CrossRef | Direct Link |
Rahman, M.A., E. Haque, M. Hasanuzzaman and I.Z. Shahid, 2011. Antinociceptive, antiinflammatory and antibacterial properties of Tamarix indica roots. Int. J. Pharmacol., 7: 527-531. CrossRef |
Ramaswamy, S., N.P. Pillai, V. Gopalkrishnan, N.S. Parmar and M.N. Ghosh, 1985. Analgesic effect of O-(β-hydroxyethyl) rutoside in mice. Indian J. Exp. Biol., 23: 219-220. PubMed | Direct Link |
Senthamarai, R., G. Umadevi and K.P.J. Gancsh, 2003. Antimicrobial activity of root extracts of Dalbergia spinosa Roxb. Ancient Sci. Life, 22: 1-3.
Trease, G.E. and W.C. Evans, 1989. Trease and Evan's Textbook of Pharmacognosy. 13th Edn., Cambridge University Press, London, Pages: 546
Whittle, B.A., 1964. The use of changes in capillary permeability in mice to distinguish between narcotic and nonnarcotic alalgesics. Br. J. Pharmacol. Chemother., 22: 246-253. PubMed | Direct Link |
Woode, E., R.A. Poku, G.K. Ainooson, E. Boakye-Gyasi, W.K.M. Abotsi, T.L. Mensah and A.K. Amoh-Barimah, 2009. An evaluation of the anti-inflammatory, antipyretic and antinociceptive effects of Ficus exasperata (Vahl) leaf extract. J. Pharmacol. Toxicol., 4: 138-151. CrossRef | Direct Link |
Zakaria, Z.A., R.N.S.R.M. Nor, M.R. Sulaiman, Z.D.F.A. Ghani, G.H. Kumar and C.A. Fatimah, 2006. Antinociceptive and anti-inflammatory properties of Melastoma malabathricum leaves chloroform extract in experimental animals. J. Pharmacol. Toxicol., 1: 337-345. CrossRef | Direct Link |
|
|
|
 |