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Antinociceptive activity of Xanthium indicum J. Koenig ex Roxb. (Asteraceae) leaves and Leucas aspera (Willd.) Link (Lamiaceae) whole plants.

Introduction

Xanthium indicum J. Koenig ex Roxb. (Asteraceae, local name: ghagra kata) is a weed commonly found in fallow lands and wild areas of Bangladesh. The roots of the plant are used in the folk medicinal system of Bangladesh for treatment of rheumatic pain. Leucas aspera (Willd.) Link (Lamiaceae, local name: dondo kolosh) is another wild herbaceous plant observed commonly throughout Bangladesh. The leaves of the plant are used by folk medicinal practitioners for treatment of headache, rheumatic pain, joint pain, pain in bones and bodyache.

Xanthium indicum has been reported to be used by the Lohit community of Arunachal Pradesh, India for treatment of inflammation-related diseases (Namsa et al., 2009). Several compounds have been isolated from the plant, which reportedly inhibited prostaglandin E(1) and prostaglandin E(2)-induced contractions in guinea pig ileum (Sadhu et al., 2003). Diterpenes, leucasperone A and leucasperosides A and B, isolated from the plant has been shown to inhibit the prostaglandin-induced contraction in guinea pig ileum (Sadhu et al., 2006). Antinociceptive and antioxidant activities have been reported for ethanolic extract of Leucas aspera roots (Rahman et al., 2007). The anti-inflammatory potential of ethanolic extract of the plant has been explored in adjuvant arthritis (Kripa et al., 2011).

Our laboratory has been investigating the bio-activities of various plants used in the folk medicines of Bangladesh for quite some time. In these studies, we have given particular attention to anti-hyperglycemic and antinociceptive activities (Anwar et al., 2010; Jahan et al., 2010; Khan et al., 2010; Mannan et al., 2010; Rahman et al., 2010; Rahman et al., 2011; Shoha et al., 2011; Sutradhar et al., 2011), for diabetes and pain are ailments affecting substantial numbers of people in all countries of the world. Since both Xanthium indicum and Leucas aspera are used by the folk medicinal practitioners for treatment of various types of pain, the objective of the present study was to evaluate the antinociceptive activity of Xanthium indicum leaves and Leucas aspera whole plants. Studies were conducted in acetic acid-induced gastric pain models in Swiss albino mice, where intraperitoneal administration of acetic acid leads to pain-induced writhings or constrictions, and antinociceptive activity of any extract or drug is demonstrated by inhibitions in the number of such writhings.

Materials and Methods

The leaves of Xanthium indicum were collected from Bogra district, Bangladesh during September, 2009. The plant was taxonomically identified at the Bangladesh National Herbarium at Dhaka (Voucher specimen No. 35,026). The air-dried leaves of Xanthium indicum were grounded into a fine powder, and 100g of the powder was extracted with methanol (1:5, w/v) for 24 hrs. The extract was evaporated to dryness. The final weight of the extract was 6.87g.

Whole plants (including flowers) of Leucas aspera were collected from Manikganj district, Bangladesh in January 2011. The plant was taxonomically identified at the Bangladesh National Herbarium at Dhaka (Voucher specimen No. 34,733). Air-dried whole plants were grounded into a fine powder, and 100g of the powder was extracted with methanol (1:5, w/v) for 24 hours. The extract was evaporated to dryness. The final weight of the extract was 8.22g.

Glacial acetic acid was obtained from Sigma Chemicals, USA; aspirin was obtained from Square Pharmaceuticals Ltd., Bangladesh.

In the present study, Swiss albino male mice, which weighed between 20-25g were used. The animals were obtained from International Centre for Diarrhoeal Disease Research, Bangladesh (ICDDR,B). Prior to experiments, all the animals were acclimatized for one week. During this one week period, mice were fed with standard mice feed obtained from ICDDR,B and water, given ad libitum. The study was conducted following approval by the Institutional Animal Ethical Committee of University of Development Alternatives, Dhaka, Bangladesh.

Antinociceptive activity of the methanol extracts were examined using previously described procedures (Shanmugasundaram and Venkataraman, 2005). Briefly, for methanolic extract of Xanthium indicum leaves, mice were divided into six groups of seven mice each. Group 1 served as control and was administered vehicle only. Group 2 was orally administered the standard antinociceptive drug aspirin at a dose of 200 mg per kg body weight. Groups 3-6 were administered methanolic leaf extract of Xanthium indicum at doses of 50, 100, 200 and 400 mg per kg body weight, respectively. For evaluating the antinociceptive activity of methanol extract of Leucas aspera whole plants, mice were divided into six groups of six mice each. Group 1 served as control and was administered vehicle only. Group 2 was orally administered the standard antinociceptive drug aspirin at a dose of 200 mg per kg body weight. Groups 3-6 were administered methanolic whole plant extract of Leucas aspera at doses of 50, 100, 200 and 400 mg per kg body weight, respectively. Following a period of 60 minutes after oral administration of standard drug or extract, all mice were intraperitoneally injected with 1% acetic acid at a dose of 10 ml per kg body weight. A period of 5 minutes was given to each animal to ensure bio-availability of acetic acid, following which period, the number of writhings was counted for 10 min.

Student's t-test was used to analyze any significant differences between control and experimental groups. P < 0.05, was considered significant as compared to control.

Results and Discussion

The results obtained from antinociceptive tests conducted with methanolic extract of Xanthium indicum leaves is shown in Table 1. The extract at doses of 50, 100, 200 and 400 mg per kg body weight demonstrated significant and dose-dependent reductions in the number of writhings. Percent inhibitions of writhings at the four afore-mentioned doses were, respectively, 31.8, 36.6, 41.2 and 42.9%. The standard antinociceptive drug, aspirin, when administered at a dose of 200 mg per kg body weight inhibited the number of writhings by 50.8%, suggesting that the antinociceptive activity of methanolic extract of Xanthium indicum leaves, although noteworthy, was not as effective as aspirin. However, although not tested in the present study, the extract may still possess an advantage over aspirin in the sense that it can avoid the side-effects associated with regular aspirin administration, like ulceration and internal bleeding.

Methanolic extract of whole plants of Leucas aspera demonstrated better antinociceptive activity in the acetic acid-induced gastric pain test. Even at the lowest dose of 50 mg extract per kg body weight, the extract reduced the number of writhings by 50.1%, which was more or less comparable to that of aspirin. The percent reduction in the number of writhings was dose-dependent and statistically significant at all four doses of the extract tested. At the higher doses of 100, 200 and 400 mg extract per kg body weight, the number of writhings was inhibited, respectively, by 51.9, 55.8 and 57.7%. The results are shown in Table 2.

Both central and peripheral analgesia can be suitably detected with the acetic acid-induced writhing test (Shanmugasundaram and Venkataraman, 2005). Peripheral analgesia involves the peripheral nociceptive afferent neuron, while central analgesia involves an afferent input thus generating a sensation of pain. Under normal circumstances, pain is associated with electrical activity in small diameter primary afferent fibers of peripheral nerves. These fibers can be non-myelinated C-fibers or fine myelinated Ad fibers. The latter causes a sensation of sharp and well-localized pain and the former causes a dull burning pain (Rang et al., 2003). Intraperitoneal administration of acetic acid (1%) leads to pain and inflammation mediated through production of prostaglandins [mainly prostacyclines (PGI2) and prostaglandin-E (PG-E)], which are reported to be responsible for excitation of the Ad-nerve fibers, leading to sensation of pain (Reynolds 1982). Therefore any agent that lowers the number of abdominal constrictions will demonstrate analgesia by inhibition of prostaglandin synthesis.

It is to be noted that several compounds have been isolated from Leucas aspera, which reportedly inhibited prostaglandin E(1) and prostaglandin E(2)-induced contractions in guinea pig ileum (Sadhu et al., 2003, 2006). These compounds may play a role in lessening the sensation of pain in acetic acid-injected mice, and so reduce the number of writhings, which are produced in response to the gastric pain felt by mice on receiving acetic acid. Whether this is the actual mechanism for the observed reduced number of writhings is currently being under investigation in our laboratory. If so, the probable mechanism can lie in the inhibition of cyclooxygenases' expression or activities, which are involved in prostaglandin synthesis. Also to be noted is the fact that Xanthium indicum is used by the Lohit community of Arunachal Pradesh, India for treatment of inflammation-related diseases (Namsa et al., 2009). Since prostaglandins are also implicated in inflammation, the underlying mechanism behind the antinociceptive activity of this plant may be the same as that for Leucas aspera. From the observed antinociceptive effects of whole plant or plant parts of the two plants evaluated, it can be concluded that both plants merit further scientific studies towards discovery of more efficacious painkilling drugs having less or no side-effects.

References

Anwar, M.M., M.A. Kalpana, B. Bhadra, S. Rahman, S. Sarker, M.H. Chowdhury and M. Rahmatullah, 2010. Antihyperglycemic activity and brine shrimp lethality studies on methanol extract of Cajanus cajan (L.) Millsp. leaves and roots. Advances in Natural and Applied Sciences, 4: 311-316.

Jahan, F.I., M.S. Hossain, A.A. Mamun, M.T. Hossain, S. Seraj, A.R. Chowdhury, Z. Khatun, N.Z. Andhi, M.H. Chowdhury and M. Rahmatullah, 2010. An evaluation of antinociceptive effect of methanol extracts of Desmodium gangeticum (L.) DC. stems and Benincasa hispida (Thunb.) Cogn. leaves on acetic acidinduced gastric pain in mice. Advances in Natural and Applied Sciences, 4: 365-369.

Khan, Z.I., B. Nahar, M.A. Jakarta, S. Rahman, M.H. Chowdhury and M. Rahmatullah, 2010. An evaluation of antihyperglycemic and antinociceptive effects of methanol extract of Cassia fistula L. (Fabaceae) leaves in Swiss albino mice. Advances in Natural and Applied Sciences, 4: 305-310.

Kripa, K.G., D. Chamundeeswari, J. Thanka, C. Uma Maheswara Reddy, 2011. Modulation of inflammatory markers by the ethanolic extract of Leucas aspera in adjuvant arthritis. Journal of Ethnopharmacology, 134: 1024-1027.

Mannan, A., H. Das, M. Rahman, J. Jesmin, A. Siddika, M. Rahman, S. Rahman, M.H. Chowdhury and M. Rahmatullah, 2010. Antihyperglycemic activity evaluation of Leucas aspera (Willd.) Link leaf and stem and Lannea coromandelica (Houtt.) Merr. bark extract in mice. Advances in Natural and Applied Sciences, 4: 385-388.

Namsa, N.D., H. Tag, M. Mandal, P. Kalita and A.K. Das, 2009. An ethnobotanical study of traditional anti inflammatory plants used by the Lohit community of Arunachal Pradesh, India. Journal of Ethnopharmacology, 125: 234-245.

Rahman, M.S., S.K. Sadhu and C.M. Hasan, 2007. Preliminary antinociceptive, antioxidant and cytotoxic activities of Leucas aspera root. Fitoterapia, 78: 552-555.

Rahman, M., A. Siddika, B. Bhadra, S. Rahman, B. Agarwala, M.H. Chowdhury and M. Rahmatullah, 2010. Antihyperglycemic activity studies on methanol extract of Petrea volubilis L. (Verbenaceae) leaves and Excoecaria agallocha L. (Euphorbiaceae) stems. Advances in Natural and Applied Sciences, 4: 361-364.

Rahman, M.M., M.N. Hasan, A.K. Das, M.T. Hossain, R. Jahan, M.A. Khatun and M. Rahmatullah, 2011. Effect of Delonix regia leaf extract on glucose tolerance in glucose-induced hyperglycemic mice. African Journal of Traditional, Complementary and Alternative Medicines, 8: 34-36.

Rang, H.P., M.M. Dale, J.M. Ritter and P.K. Moore, 2003. Pharmacology, Churchill Livingstone, 5th Edition; pp: 562-563.

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Sadhu, S.K., E. Okuyama, H. Fujimoto and M. Ishibashi, 2003. Separation of Leucas aspera, a medicinal plant of Bangladesh, guided by prostaglandin inhibitory and antioxidant activities. Chemical & Pharmaceutical Bulletin, 51: 595-598.

Sadhu, S.K., E. Okuyama, H. Fujimoto and M. Ishibashi, 2006. Diterpenes from Leucas aspera inhibiting prostaglandin-induced contractions. Journal of Natural Products, 69: 988-994.

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Shoha, J., H. Jahan, A.A. Mamun, M.T. Hossain, S. Ahmed, M.M. Hossain, S. Rahman, R. Jahan and M. Rahmatullah, 2011. Antihyperglycemic and antinociceptive effects of Curcuma zedoaria (Christm.) Roscoe leaf extract in Swiss albino mice. Advances in Natural and Applied Sciences (in press).

Sutradhar, B.K., M.J. Islam, M.A. Shoyeb, H.N. Khaleque, M. Sintaha, F.A. Noor, W. Newaz and M. Rahmatullah, 2011. An evaluation of antihyperglycemic and antinociceptive effects of crude methanol extract of Coccinia grandis (L.) J. Voigt. (Cucurbitaceae) leaves in Swiss albino mice. Advances in Natural and Applied Sciences (in press).

Corresponding Author: Professor Dr. Mohammed Rahmatullah, Pro-Vice Chancellor and Dean, Faculty of Life Sciences University of Development Alternative House No. 78, Road No. 11A (new) Dhanmondi, Dhaka-1205 Bangladesh Telephone: +88-01715032621 Fax: +88-02-815739 E-mail: rahamatm@hotmail.com

A.N.M. Farhad Hossan, Farhana Zaman, Mukta Rani Barman, Safia Khatoon, Marzia Zaman, Fatema Khatun, Tamim Mosaiab, Fariha Mostafa, Mariz Sintaha, Farhana Jamal, Mohammed Rahmatullah

Faculty of Life Sciences, University of Development Alternative, Dhanmondi, Dhaka-1205, Bangladesh.
Table 1: Antinociceptive effect of crude methanol extract of Xanthium
indicum leaves in the acetic acid-induced gastric pain model mice.

                    Dose (mg/kg      Mean number
Treatment           body weight)     of writhings       % inhibition

Control (Group 1)       10 ml      9.00 [+ or -] 0.65      --
Aspirin (Group 2)      200 mg      4.43 [+ or -] 1.09      50.8 *
Xanthium indicum        50 mg      6.14 [+ or -] 1.26      31.8 *
(Group 3)
Xanthium indicum       100 mg      5.71 [+ or -] 1.06      36.6 *
(Group 4)
Xanthium indicum       200 mg      5.29 [+ or -] 0.78      41.2 *
(Group 5)
Xanthium indicum       400 mg      5.14 [+ or -] 1.03      42.9 *
(Group 6)

All administrations (aspirin and extract) were made orally. Values
represented as mean [+ or -] SEM, (n=7); * P < 0.05; significant
compared to control.

Table 2: Antinociceptive effect of crude methanol extract of Leucas
aspera whole plants in the acetic acid-induced gastric pain model
mice.
                    Dose (mg/kg       Mean number
Treatment           body weight)      of writhing       % inhibition

Control (Group 1)       10 ml      8.50 [+ or -] 0.88      --
Aspirin (Group 2)      200 mg      4.17 [+ or -] 0.79      50.9 *
Leucas aspera           50 mg      4.33 [+ or -] 0.42      50.1 *
(Group 3)
Leucas aspera          100 mg      4.17 [+ or -] 0.75      51.9 *
(Group 4)
Leucas aspera          200 mg      3.83 [+ or -] 0.64      55.8 *
(Group 5)
Leucas aspera          400 mg      3.67 [+ or -] 0.33      57.7 *
(Group 6)

All administrations (aspirin and extract) were made orally. Values
represented as mean [+ or -] SEM, (n=6); * P < 0.05; significant
compared to control.
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Title Annotation:Original Article
Author:Hossan, A.N.M. Farhad; Zaman, Farhana; Barman, Mukta Rani; Khatoon, Safia; Zaman, Marzia; Khatun, Fa
Publication:Advances in Natural and Applied Sciences
Article Type:Report
Geographic Code:9BANG
Date:Apr 1, 2011
Words:2416
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