Antinociceptive Effect of Methanol Extract of Dalbergia Sissoo Leaves in Mice Md

Mannan et al. BMC Complementary and Alternative Medicine (2017) 17:72 DOI 10.1186/s12906-017-1565-y RESEARCH ARTICLE Open Access Antinociceptive effect of methanol extract of Dalbergia sissoo leaves in mice Md. Abdul Mannan*, Ambia Khatun and Md. Farhad Hossen Khan Abstract Background: Dalbergia sissoo DC. (Family: Fabaceae) is a medium to large deciduous tree, is locally called “shishu” in Bangladesh. It is used to treat sore throats, dysentery, syphilis, bronchitis, inflammations, infections, hernia, skin diseases, and gonorrhea. This study evaluated the antinociceptive effect of the methanol extract of D. sissoo leaves (MEDS) in mice. Methods: The extract was assessed for antinociceptive activity using chemical and heat induced pain models such as hot plate, tail immersion, acetic acid-induced writhing, formalin, glutamate, and cinnamaldehyde test models in mice at the doses of 100, 200, and 400 mg/kg (p.o.) respectively. Morphine sulphate (5 mg/kg, i.p.) and diclofenac sodium (10 mg/kg, i.p.) were used as reference analgesic drugs. To confirm the possible involvement of opioid receptor in the central antinociceptive effect of MEDS, naloxone was used to antagonize the effect. Results: MEDS demonstrated potent and dose-dependent antinociceptive activity in all the chemical and heat induced mice models (p < 0.001). The findings of this study indicate that the involvement of both peripheral and central antinociceptive mechanisms. The use of naloxone verified the association of opioid receptors in the central antinociceptive effect. Conclusions: This study indicated the peripheral and central antinociceptive activity of the leaves of D. sissoo. These results support the traditional use of this plant in different painful conditions. Keywords: Dalbergia sissoo, Antinociceptive, Opioid receptors Background filtrate is used to wash hair for removing dandruff and Dalbergia sissoo DC. (Family: Fabaceae) is a medium to for long hair [10]. large deciduous tree, is commonly known as sisu, shee- Isoflavones, biochanin-A, muningin, sissotrin, amyrin, sham, tahli, tali, jag at different parts of the world and is stigmasterol have been isolated from the aerial parts of locally called “shishu” in Bangladesh. It is usually used to Dalbergia sissoo Roxb [11]. The stem-bark has been treat sore throats, dysentery, syphilis, and gonorrhoea reported to contain dalberjenone, dalbergin and methyl [1]. It is used in conditions like blood purifier, leprosy, dalbergin and a new 4-phenyl chromene, dalbergichro- headaches, bronchitis, inflammations, infections, hernia, mene [12]. The mature pods contain a new isoflavone and skin diseases [2]. It has been also reported to use as glucoside identified as caviunin 7-0-gentiobioside [13], stimulant, astringent [3]; colorectal cancer and bacterial isoflavone diglucoside recognize as isocaviunin 7- infections [4]. Juice of the leaves is useful for anthelmin- 0-gentiobioside [14]. The root bark of Dalbergia sissoo tic, eye and nose diseases. It also used in scabies, contain a chalcone, 2, 3- dimethoxy-40-g, g- burning sensation, scalding urine and digestive disorders dimethylallyloxy-20-hydroxychalcone; an isoflavone, 7-g, [5–8]. It is used as expectorant, anti-emetic, leucoderma, g-dimethylallyloxy- 5-hydroxy-40-methoxyisoflavone; a ulcers, and gout like medicinal properties [9]. Boiled leaf flavone, 7-hydroxy-6-methoxyflavon; a isoflavone, biochanin A and rotenoid, dehydroamorphigenin [15]. Rhamnose, galactose, and glucuronic acid have been isolated from the leave [16]. Compounds obtained from * Correspondence: [email protected] Department of Pharmacy, Stamford University Bangladesh, 51, Siddeswari Dalbergia sissoo like an isoflavone, biochanin is a potent Road, Dhaka 1217, Bangladesh © The Author(s). 2017 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. Mannan et al. BMC Complementary and Alternative Medicine (2017) 17:72 Page 2 of 13 chemotherapeutic cancer preventive agent with a dis- Equipment Co. Ltd.) and 17.59 g extract (Yield 5.86%) tinct estrogenic activity has been isolated from the fresh was obtained. This extract was used for the acute flowers of Dalbergia sissoo. Two rare glycosides such as toxicity and antinociceptive activity studies. kaempferol and quercetin rutinosides were isolated in a low yield from the flowers of Dalbergia sissoo [17, 18]. Animals Pharmacological studies have evaluated the analgesic Swiss albino mice (20–25 g) were collected from the and anti-pyretic [19], anti-diabetic [20], anti-inflammatory Animal Research Branch of the International Center for [21] and anti-diarrheal effect of the extract [22]. The plant Diarrheal Disease and Research, Bangladesh (ICDDR, B). bark has been reported the anti-nociceptive [23], anti- Animals were kept under standard laboratory conditions oxidant [24] and anti-spermatogenic activity [25]. The oil (room temperature: 25.0 ± 2.0 °C, relative humidity: 55– extracted from wood scrapings of Dalbergia sissoo has 65% and 12 h light/dark cycle) with food and water dur- shown dose dependent larvicidal activity against mosqui- ing the adaptation period. The animals were acclimated toes [26]. Anti-osteogenic activity has also been performed to the laboratory environment for a period of two weeks this crude extract of Dalbergia sissoo [27]. The plant ex- prior to performing the experiments. Mice were fasted tract has been used as an alternative to synthetic pesticides overnight prior to the experiments. All the experimental for termite control in buildings [28]. The seeds extract has animals were treated following the Ethical Principles and modest analgesic and notable anti-pyretic activities [29]. Guidelines for Scientific Experiments on Animals (1995) The ethanolic extract of the bark of Dalbergia sissoo Roxb. formulated by The Swiss Academy of Medical Sciences was investigated for its activity against helminthic infec- and the Swiss Academy of Sciences. The Ethics Commit- tions [30]. In the search for the molluscicidal activity, the tee of Stamford University Bangladesh (SUB/IAEC/ plant extracts were evaluated against egg masses and 16.01) approved all experimental protocols. adults of Biomphalaria pfeifferi and the snail intermediate host of Schistosoma manson in Nigeria [31]. In-vivo, Drugs and treatments intrastriatal injection of mitochondrial toxin in rats pro- The control group orally received deionized water at the duce excitotoxic lesions, while chronic administration of volume of 0.1 mL/mouse 30 min before the experiments. 3-NP produces excitotoxic-like lesions regionally re- The positive control group intraperitoneally (i.p.) stricted to striatum in both rats and non-human primates received standard drug morphine in hot plate, and tail [32]. It prevents central nervous system damage [33]. immersion test at the dose of 5 mg/kg and diclofenac According to the previous study, leaves and barks sodium in acetic acid-induced writhing, formalin- of D. sissoo presented the antinociceptive properties induced licking, glutamate-induced paw licking, and [19, 23]. Based on the studies, we designed and con- cinnamaldehyde-induced licking test at the dose of ducted the present work to assess the effect of methanol 10 mg/kg 15 min before the experiments. MEDS was ad- extract of D. sissoo leaves with six pain models in mice ministered orally at the doses of 100, 200, and 400 mg/ and two mechanism of action study for possible activity. kg (b.w.) 30 min before the experiments. To evaluate the The present research also warrants further investigation involvement of opioid-mediated antinociceptive activity, into the complex mechanism of action of methanol naloxone was administered (i.p.) at the dose of 2 mg/kg extract of D. sissoo leaves on induced antinociception. 15 min before morphine sulfate (5 mg/kg, i.p.) or MEDS (100, 200, and 400 mg/kg, p.o.) administration in the hot Methods plate and tail immersion test. Methylene blue (20 mg/ Plant material and extraction kg) and glibenclamide (10 mg/kg) were intraperitoneally The fresh leaves of D. sissoo were collected from Khan- employed 15 min before control (Equivalent volume of pur, Bogra, Bangladesh in the month of June 2015. The deionized water, 0.1 mL/mouse, p.o.), or MEDS (100, plant samples were then identified and authenticated by 200, and 400 mg/kg, p.o.) administration to evaluate the Sarder Nasir Uddin, Senior Scientific Officer, Bangladesh involvement of cyclic guanosine monophosphate National Herbarium, Dhaka, Bangladesh. A voucher spe- (cGMP) and ATP-sensitive K+ channel pathway respect- cimen number (DACB: 42317) has already been depos- ively. All the doses of drugs and MEDS were prepared ited in the Herbarium for further reference. The fresh using deionized water. leaves were dark dried for one week and grinded to a fine powder. Powdered dried leaves (300 g) were macer- Acute toxicity test ated with 1000 mL of methanol with occasional stirring Acute toxicity studies were carried on mice according to at 25 ± 2 °C for 3 days in a beaker. The extract was then the method proposed by Ghosh, MEDS at the doses of 50, filtered using a Whatman No. 1 filter paper and a steril- 100, 300, 1000, and 3000 mg/kg body weight were orally ized cotton filter. The solvent was completely removed administered to separate groups of the mice (n =5)after by rotary evaporator (BC-R 201 Shanghai Biochemical overnight fasting. Subsequent to administration of MEDS, Mannan et al. BMC Complementary and Alternative Medicine (2017) 17:72 Page 3 of 13 the mice observed closely for the first 3 h for toxic mani- and MEDS. Then, the % MPE was calculated from using festations like enlarged motor activity, salvation, convul- the same formula used in hot plate test.

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